How To Streamline Diagnostic Development and Manufacturing

Diagnostics guide Accelerate molecular and immunodiagnostic test development and manufacture 2 Diagnostics have always played a critical role in treating illnesses and managing health. The ability to gain a fast, accurate understanding of a person’s disease status helps to drive effective treatment in a timely manner and improve patient outcomes. With new, more targeted, and personalized therapies coming to the market quickly and changing the way we treat complex conditions, it’s increasingly important that diagnostics developers and manufacturers keep pace. Technological advancements, along with recent industry shifts and learnings, are setting the stage for exciting changes in diagnostics and clinical care. Introduction 3 Lab-based molecular diagnostics Lab-based immunodiagnostics Point-of-care molecular diagnostics Point-of-care immunodiagnostics Diagnostic services With our diagnostics services and our extensive portfolio of magnetic beads, membranes, and paper, we aim to be at the forefront of technological development to support molecular and immunodiagnostic workflows for tests used in a lab or a point-of-care (POC) environment. With technical expertise and extensive manufacturing capabilities supported by security of supply commitments, we provide solutions, backed by validated protocols, which minimize disruption to your existing workflows and help you commercialize your assay faster. Explore our range of services: 4 Lab-based molecular diagnostics Sample collection Nucleic acid extraction/isolation Amplification and enrichment Detection Process Sample collected e.g. by blood draw, swab or collection of other body fluid into stabilization matrix to preserve content Sample is prepared and nucleic acid is isolated Specific sequences are amplified with primers/probes or library preparation for NGS Detection via digital droplet PCR (ddPCR), quantitative PCR (qPCR), LAMP readout or by NGS Cytiva products • 903 card • SeraSil-Mag™ silica beads • Sera-Mag™ carboxyl-modified magnetic beads and speedbeads • Sera-Mag oligo (dT) magnetic beads • Sera-Xtracta™ HMW DNA kit • Sera-Xtracta cell-free DNA kit • Sera-Xtracta virus/pathogen kit • Various membranes and pads may also be an option here • Nucleotides • Taq DNA polymerase • Sera-Mag select size selection and PCR clean-up reagent • Sera-Mag carboxyl-modified magnetic beads and speedbeads • Sera-Mag streptavidin magnetic beads • Sera-Mag streptavidin-blocked magnetic beads Cytiva services Custom bead formats/types | Custom bead conjugations Custom reagents Lyo-Stable™ lyophilization service | Contract manufacturing Click anywhere in the table to explore 5 Sample collection Development of lab-based assays for molecular diagnostics Molecular diagnostics utilizes the techniques of molecular biology to analyze biological markers in an individual’s genome to understand how the cells express their genes as proteins. Molecular diagnostics is used to predict and diagnose disease, select treatments, and monitor the effectiveness of therapies. Fluorescence in situ hybridization (FISH), quantitative PCR (qPCR), microarrays, and next-generation sequencing (NGS) are the main technologies for studying genomic abnormalities in clinical applications. Only microarrays and NGS are designed to look at multiple areas of the genome simultaneously. Developing solutions for molecular diagnostics that work well with the analytical technologies available is an area of key focus for us. The increasing availability of genome sequencing has been a key factor for the growth of molecular diagnostics. Genome sequencing allows disease diagnosis at an earlier stage, and with the discovery of relevant biomarkers, it enables more targeted therapeutic interventions. The trend toward a more personalized approach to healthcare is therefore another driver for the expanding importance of molecular diagnostics. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 6 Whatman™ 903 proteinsaver card Whatman™ 903 proteinsaver card is a 903-sample protein collection card with five half-inch sample spots for protein collection, transport, and storage. Ready-to-use cards and kits save time and costs during protein collection, transport, and archiving. The proteinsaver card has a wraparound cover with spaces for name and date of collection. The card fits into our Whatman foil barrier resealable bags for storage. The card is imprinted with the universal biohazard symbol in accordance with USP regulations. Features and benefits Storage: each circle holds 75 to 80 µL of sample Strict quality control and ISO9001 manufacturing standards: ensures high quality, reproducibility, and purity We can design customized components and solutions to help you build smarter diagnostic assays using Whatman papers and filters. Product Product code 903 proteinsaver cCard (EU) 10531018 903 proteinsaver cCard (US) 10534612 Application note Use of the Sera-Xtracta™ virus/pathogen kit for purification of SARS-CoV-2 samples collected on Whatman 903 proteinsaver cards. Learn more Learn more about 903 proteinsaver card. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 7 Nucleic acid extraction and isolation Magnetic beads: a simple guide Magnetic beads are made up of tiny particles of iron oxides, such as magnetite (Fe3O4), which give the beads superparamagnetic properties. Superparamagnetic beads are different from ferromagnets in that they exhibit magnetic behavior only in the presence of an external magnetic field. This property is dependent on the small size of the particles in the beads and enables the beads to be separated in suspension, along with anything they are bound to. Because they don’t attract each other outside of a magnetic field, they can be used without any concern about unwanted clumping. Many types of magnetic beads are available. Different surface coatings and chemistries give each type of bead its own binding properties, which can be used for magnetic separation (isolation and purification) of nucleic acids, proteins, or other biomolecules in an easy, effective, and scalable way. This ease-of-use makes magnetic beads automation friendly and well suited for a range of applications including sample preparation for NGS and PCR, protein purification, molecular and immunodiagnostics, and magnetic-activated cell sorting (MACS), and many others. They also ease some of the challenges associated with extracting nucleic acids from different sample types. 7 Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 8 Type Properties Applications Variations Carboxylate-modified magnetic beads • Can associate with nucleic acids for direct capture • Surface suitable for conjugation through covalent bonding • Can capture molecules containing amino groups Conjugation or direct binding applications: • Covalent attachment • Affinity purification and pull-down • Nucleic acid isolation and purification • NGS size selection High-speed version available Amine-blocked magnetic beads Surface suitable for conjugation through covalent bonding • Nonsurfactant, nonprotein-blocked surface • Low nonspecific binding Conjugation applications similar to carboxylate-modified beads High-speed version available Oligo (dT)-coated magnetic beads Hybridizes with mRNA poly-A tails • High colloidal stability mRNA binding applications: • mRNA extraction and purification • RT-PCR • cDNA library construction • Subtractive hybridization • NGS (RNA sequencing) Streptavidin-coated magnetic beads Binds biotinylated ligands such as proteins, nucleic acids, and peptides • Covalently bound streptavidin coating • Fast reaction kinetics • Low nonspecific binding • High throughput and precise Immunoassay and molecular biology applications: • Sample preparation and assay development for genomics and proteomics High-speed version available Biotin binding ranges: • 2500 to 3500 pmol/mg • 3500 to 4500 pmol/mg • 4500 to 5500 pmol/mg Streptavidin-blocked magnetic beads • Binds biotinylated ligands such as proteins, nucleic acids, and peptides • Nonsurfactant, nonprotein-blocked surface • Lower nonspecific binding than streptavidin-coated beads via additional blocking of nonspecific binding sites High-specificity biotin binding applications: • Molecular and immunodiagnostics • NGS library preparation High-speed version available NeutrAvidin-coated magnetic beads • Binds biotinylated ligands such as proteins, nucleic acids, and peptides • Nonsurfactant, nonprotein-blocked surface • Lower nonspecific binding than streptavidin-coated beads via additional blocking of nonspecific binding sites Alternative to streptavidin in immunoassay and molecular biology applications • Sample preparation and assay development for genomics and proteomics High-speed version available Biotin binding range: • 3500 to 4500 pmol/mg Protein A/G magnetic beads • Binds IgA and IgG proteins • Coating based on IgA/IgG fusion protein • Broad binding capabilities Antibody isolation applications: • Affinity purification and pull-down • Immunoprecipitation Silica-coated magnetic beads • Reversibly binds nucleic acids based on salt concentration • Monodisperse particles with narrow size ranges of 400 µm or 700 µm Applications with low sample amounts: • Nucleic acid extraction for molecular diagnostics applications such as qPCR Mag Sepharose™ beads • Broad range of ligand options • Porous, providing greater surface area than other magnetic beads Convenient alternative to Sepharose columns, with protein purification applications including: • Affinity purification or capture • Immunoprecipitation Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 9 Product Quantity Product code SeraSil-Mag 400 nm magnetic particles 5 mL 29357369 SeraSil-Mag 400 nm magnetic particles 60 mL 29357371 SeraSil-Mag 400 nm magnetic particles 450 mL 29357372 SeraSil-Mag 700 nm magnetic particles 5 mL 29357373 SeraSil-Mag 700 nm magnetic particles 60 mL 29357374 SeraSil-Mag 700 nm magnetic particles 450 mL 29357375 Explore Explore ways to optimize your magnetic bead workflow in ‘The scientist’s guide to magnetic beads’. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection SeraSil-Mag™ silica beads For use in Sanger sequencing and NGS SeraSil-Mag™ silica coated superparamagnetic beads deliver highpurity extraction solutions for highly sensitive applications when sample is scarce. The beads provide an optimal binding surface with regular morphology to optimize binding efficiency and reduce variability, simplifying the transition from column purification to bead-based purification. Optimized to isolate highly purified DNA and RNA for downstream applications such as qPCR or sequencing. Features and benefits High iron oxide content (60 emu/g): Fast magnetic response (~ 5 s) shortens time of magnetic steps during isolation Uniformity: Particles are uniform in size (submicroscale diameter 700 nm and 400 nm [monodispersed]), providing narrow size distribution Low sedimentation rate: Good buoyancy enhances ease of handling, automation, and reproducibility Purity: Used to isolate and purify genomic DNA from whole human blood providing A260/A280 ratios between 1.70 to 1.90 and A260/A230 ratios as high as 2is a hydrophilic, fibrous matrix designed for use in procedures requiring the isolation of leukocytes from whole blood. 10 Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Product Quantity Product code Sera-Mag carboxylate-modified [E7] magnetic particles 15 mL 24152105050250 Sera-Mag carboxylate-modified [E7] magnetic particles 100 mL 24152105050350 Sera-Mag carboxylate-modified [E7] magnetic particles 1000 mL 24152105050450 Sera-Mag carboxylate-modified [E3] magnetic particles 15 mL 44152105050250 Sera-Mag carboxylate-modified [E3] magnetic particles 100 mL 44152105050350 Sera-Mag carboxylate-modified [E3] magnetic particles 1000 mL 44152105050450 Explore Explore ways to optimize your magnetic bead workflow in ‘The scientist’s guide to magnetic beads’. Sera-Mag™ carboxylate beads and speedbeads For use in Sanger sequencing and NGS Carboxylic groups on the surface of Sera-Mag speedbeads and SeraMag carboxylate-modified magnetic beads permit easy covalent coupling to target biomolecules of interest, such as proteins and nucleic acids, using convenient carbodiimide chemistry. The cauliflower-shaped surface, paired with proprietary Sera-Mag beads and speedbead chemistry, provides a large surface area and offers excellent sensitivity and low nonspecific binding for greater accuracy. This large surface area can maximize sample retention or reduce the amount of beads required. E3 and E7 refer to the different manufacturing process for the beads. E3 and E7 beads behave similarly, and we continue to provide both bead types to our customers for test and validation in their chosen application. Sera-Mag carboxylate beads Sera-Mag carboxylate-modified magnetic beads combine a fast magnetic response time and high binding capacity, sensitivity, stability, and physical integrity. Features and benefits Ease of use: Covalent coupling of proteins, nucleic acids, etc. to carboxyl groups on the surface using standard coupling technologies Convenient: Isolation, selection, and clean-up of nucleic acids or direct conjugation of specific oligos and enzymes 11 Request a sample Request a sample of magnetic beads. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Product Quantity Product code Sera-Mag speedbead carboxylate-modified [E7] magnetic particle 15 mL 45152105050250 Sera-Mag speedbead carboxylate-modified [E7] magnetic particle 100 mL 45152105050350 Sera-Mag speedbead carboxylate-modified [E7] magnetic particle 1000 mL 45152105050450 Sera-Mag speedbead carboxylate-modified [E3] magnetic particle 15 mL 65152105050250 Sera-Mag speedbead carboxylate-modified [E3] magnetic particle 100 mL 65152105050350 Sera-Mag speedbead carboxylate-modified [E3] magnetic particle 1000 mL 65152105050450 Sera-Mag speedbead carboxylate-modified magnetic particles Sera-Mag speedbeads have a second layer of magnetite applied through the same core shell design process. This second layer allows a reaction twice as fast as the Sera-Mag carboxylatemodified beads when in the presence of a magnetic field. Speedbeads are especially useful when the reaction medium is highly viscous, and in clinical assays requiring a faster magnetic response time. Features and benefits Convenient: Isolation, selection, and clean-up of nucleic acids or direct conjugation of specific oligos and enzymes Reliable: Fast, precise, and high binding capacity for sample preparation, nucleic acid isolation, proteomics, and immunoassay applications 12 Product Quantity Product code Sera-Mag oligo (dT) coated magnetic particles 1 mL 38152103011150 Sera-Mag oligo (dT) coated magnetic particles 5 mL 38152103010150 Sera-Mag oligo (dT) coated magnetic particles 100 mL 38152103010350 12 Explore Download our troubleshooting guide on magnetic bead characteristics. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Sera-Mag oligo (dT) coated magnetic particles For use in reverse transcriptase qPCR (RT-qPCR), microarrays, Sanger sequencing, and NGS Colloidally stable Sera-Mag oligo (dT) coated magnetic particles contain covalently bound oligo (dT)14 and remain in suspension for extended periods of time in the absence of a magnetic field, making them well suited for capturing or isolating mRNA from a variety of sources. Oligo (dT) particles can also be used as a universal base particle for coupling unique oligo sequences. Simply synthesize the oligo with a poly-A tail for easy attachment to the oligo (dT) particles. Features and benefits Versatile: Once isolated, selective purification of mRNA from total RNA for NGS, RT-PCR, cDNA library construction, or subtractive hybridization can be performed Performance: The approximate mRNA binding-capacity is 11 µg of mRNA per mg of particles (dependent upon sample and message length) Customizable: Custom oligo (dT) available for more specific sample focus binding 13 Sera-Xtracta™ high-molecular-weight (HMW) DNA kit High yield extraction of high-molecular-weight genomic DNA from blood, buffy coat, saliva, cultured mammalian cells, and solid tissue with a simplified protocol. Sera-Xtracta™ HMW DNA kit* offers magnetic bead-based extraction and purification of genomic DNA from whole blood (treated with EDTA, citrate, or heparin), buffy coat, saliva, cultured mammalian cells, and solid tissue samples. The extraction protocols are designed to efficiently remove contaminants and PCR inhibitors while minimizing shearing, resulting in high-quality, high-molecular-weight genomic DNA with minimal RNA carryover. Sera-Xtracta HMW DNA kit* has been optimized for expanded applications and is a direct replacement for the Sera-Xtracta genomic DNA kit*. The components of the kit and the product code remain the same. Please contact Scientific Support for instructions for use (IFU), the protocol, and the datafile for Sera-Xtracta genomic DNA kit*. Features and benefits Suitable for long-read sequencing: Isolates high-molecularweight DNA, > 200 kb Efficient scale-up and flexibility: Provides robust chemistry for sample input volumes from 200 μL to 2 mL Efficient removal of inhibitors to give high-purity genomic DNA: Typical purity ratio (A260/A280 and A260/A230) of > 1.7 Automation friendly reagents and easy to use protocols Simplified protocol minimizes the copurification of RNA Compatible with molecular biology techniques, including NGS, cloning, restriction enzyme digestion, PCR amplification, and genotyping applications The kit uses chaotropic agents to extract DNA from samples, denature protein components, and promote the selective binding of DNA to the silica-coated magnetic beads. Proteinase K is the protease of choice to digest protein from samples because it is active even when enzyme inhibitors such as EDTA and detergents are present in samples. Denatured contaminants are easily removed by subsequent washing of the silica beads with an ethanolic buffer set. The purified genomic DNA is eluted in a low ionic strength buffer at a concentration suitable for most downstream molecular biology applications. Automation scripts are available; contact Scientific Support. Product Quantity Product code Sera-Xtracta HMW DNA kit* 96 purifications 29429140 *For research use only (RUO). Not for diagnostic use. Request a sample Request a sample of magnetic beads. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 14 Product Quantity Product code Sera-Xtracta cell-free DNA kit* 96 purifications 29437807 14 *For research use only Application note Detection of cancer-associated mutations in liquid biopsies for the identification of therapeutic targets. Application note Targeted NGS biomarker discovery using serum from stage IV non-small cell lung cancer (NSCLC) and colorectal cancer (CLC) patient samples. Application note Plasma based-monitoring of therapeutic response and resistance in patients positive for epidermal growth factor receptor (EGFR) biomarker. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Sera-Xtracta cell-free DNA kit For use in RT-qPCR, qPCR, and NGS Designed to select for small-fragment cell-free DNA (cfDNA) while minimizing genomic DNA contamination, Sera-Xtracta cell-free DNA kit* offers efficient extraction and purification of cfDNA from plasma. High yield and sensitivity make the kit well-suited for applications where sample is precious such as cancer diagnosis and monitoring. Features and benefits High yield and sensitivity: High recovery of small-fragment cfDNA in the size range of 50 to 300 bp, which allows capture of more cfDNA and increased sensitivity levels. Minimal copurification of higher molecular weight genomic DNA is required due to active selection of smaller fragments, which further improves sensitivity in downstream applications. Scalability: One kit covers 0.5 mL to 4 mL input of plasma. Efficient: Procedure can be completed in less than 120 minutes. Ease of use: Compatible with molecular biology techniques including NGS; qPCR; droplet digital PCR (ddPCR); beads, emulsion, amplification, magnetics (BEAMing); and other amplification and genotyping applications. 15 Product Quantity Product code Sera-Xtracta virus/pathogen kit* 96 purifications 29506009 Sera-Xtracta virus/pathogen kit* 1000 purifications 29514201 Membranes and pads Various membranes, pads, and paper may be used for nucleic acid isolation. Please refer to the section on molecular assay development in the point-of-care section. Request a sample Request a sample of magnetic bead based kits. 15 *For research use only Application note Sensitive detection of total nucleic acid (RNA and DNA) from swabs, blood and transport media using Sera-Xtracta virus/pathogen kit. Application note Use of the Sera-Xtracta virus/pathogen kit for purification of SARS-CoV-2 samples collected on Whatman 903 proteinsaver cards. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Sera-Xtracta virus/pathogen kit For use in RT-qPCR and qPCR Sera-Xtracta virus/pathogen kit* is designed for high-throughput total nucleic acid (DNA/RNA) isolation from bacteria and viruses including adenovirus (Type 14), influenza A (H3N2), and COVID-19. For use with respiratory biological matrices, blood, and universal transport media, the kit provides a simple and rapid method to optimize the workflow for sensitive detection of viruses and other pathogens found in low concentrations. Features and benefits Reproducible yields: Offers the advantages of solid phase extraction (using magnetic beads) Simplified protocol: Can be adapted for both manual and automated high-throughput processing Scalability: Up to 400 µL of sample Rapid: Extraction procedure can be completed in less than 30 minutes. Ease of use: Compatible with molecular biology techniques, including qPCR, RT-qPCR, ddPCR, and NGS 16 Custom bead conjugation Even with all these surface chemistry options, it’s impossible to cover every need and eventuality. That’s where custom conjugation comes in. Do you need to conjugate a custom ligand? Or need a custom particle size? We make it possible to customize all our Sera-Mag™ magnetic beads. Our dedicated customization experts can help you every step of the way, from defining the product specifications to delivery completion. We provide custom conjugations of enzymes or antibodies, as well as a range of custom ligands that we can develop in parallel with your projects. We also offer lyophilization of the customized microspheres as part of our Lyo-Stable™ services, based on stabilization technology. Drawing on our R&D and manufacturing knowledge and history of supplying magnetic beads to kit manufacturers, we will provide custom magnetic bead technology that fit your specifications. From completing complex conjugations, to performing your quality control tests before the beads leave our factory, we are equipped to meet your needs. Read more Read more about magnetic bead conjugation in our ‘A magnetic bead for every need’ knowledge article. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 17 Amplification method PCR MDA Key features • Requires thermal cycling. • Suited to small and medium size amplicons. • Requires knowledge of target sequence. • Uses specific primers. • Product requires purification for downstream analysis. • Isothermal reaction (30°C). • Suited to large amplicons. • Requires no knowledge of target sequence. • Uses random hexamer primers. • Product requires no further purification. • Minimal hands-on time. Application examples • Targeted enrichment in NGS. • Quantitative analysis (e.g. qPCR). • Whole genome amplification. • Genotyping. • Site-directed mutagenesis. • Direct amplification of circular vectors in cell-free cloning. • cDNA amplification. • Whole genome amplification. • SNP identification. Table 1: Comparison of PCR and MDA approaches As workflow for MDA is also simpler than PCR, requiring no thermal cycling, commercial kits offer straightforward one-tube, one-temperature formats that facilitate automation and high-throughput sample amplification. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Amplification and enrichment While isothermal amplification using Phi29 has several benefits over traditional polymerase chain reaction (PCR), it is not a replacement for the old workhorse. PCR remains a key component of many molecular biology workflows, such as sequencing, genotyping, and site-directed mutagenesis. For situations with amplicons within the reliable limit of highfidelity polymerases (usually less than 10 kbp) and known sequences (or at least primer binding sites), PCR remains the first choice. In other situations, whole genome amplification (WGA) for singlecell sequencing for example, you have more options. There are PCR methodologies for WGA, but they are relatively complex, potentially less robust, and require more optimization than multiple displacement amplification (MDA). Using Phi29-based MDA could provide a faithful and uniform amplification of the original genome with fewer variables, larger amplicons, and less effort than PCR. But during library prep, you are again likely to return to PCR for steps like end-repair and adding functional elements to your fragments. Beyond sequencing, however, the features of Phi29 DNA polymerase also lend themselves well to several other applications (Table 1). For example, the high fidelity, and therefore low rates of false positives and negatives, make Phi29 well suited for identifying single nucleotide polymorphisms (SNPs) and other mutations. 18 Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Product Quantity Product code Amersham dATP, solution, 100 mM 25 µmol 28406501 Amersham dATP, solution, 100 mM 100 mmol 28406502 Amersham dATP, solution, 100 mM 500 µmol 28406503 Amersham dATP, solution, 100 mM 100 µmol 28406512 Amersham dATP, solution, 100 mM 100 µmol 28406522 Amersham dATP, solution, 100 mM 25 mmol 28406531 Amersham dATP, solution, 100 mM 100 µmol 28406532 Amersham dATP, solution, 100 mM 25 µmol 28406541 Amersham dATP, solution, 100 mM 100 µmol 28406542 Amersham dNTP set (100 mM each A, C, G, T) 4 × 25 µmol 28406551 Amersham dNTP set (100 mM each A, C, G, T) 4 × 100 µmol 28406552 Amersham dNTP set (100 mM each A, C, G, T) 4 × 500 µmol 28406553 Amersham dNTP set (100 mM each A, C, G, T) 10 mmol 28406557 Amersham dNTP set (100 mM each A, C, G, T) 4 × 10 µmol 28406558 Nucleotides For use in FISH, RT-qPCR, qPCR, microarrays, Sanger sequencing, and NGS Amersham™ dNTPs are high-purity deoxynucleotides for amplification, dideoxy sequencing, labeling, mutagenesis, cDNA synthesis, and expression profiling. They are free from DNase, RNase, and nicking enzyme activity. Features and benefits Performance: Greater than 99% triphosphate purity (by HPLC) for consistency Convenient: Buffer-free, ready-to-use solutions at a variety of concentrations Tested: Functionally tested to produce a 20.7 kb PCR amplification product from λ-DNA Also available: high-purity rNTPs providing easy-to-use solutions that save time for in vitro transcription. All rNTPs are lot tested for ribonuclease contamination. 19 Product Quantity Product code Taq DNA Polymerase (cloned) 250 27079804 Taq DNA Polymerase (cloned) 4 × 250 27079805 Taq DNA Polymerase (cloned) 10 × 250 27079806 Taq DNA Polymerase (cloned) 25 000 28937348 19 Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Taq DNA polymerase For use in RT-qPCR, qPCR, microarrays, Sanger sequencing, and NGS Cloned Taq DNA polymerase is the recombinant protein form of the native enzyme from Thermus aquaticus expressed in E. coli. Like native Taq, it polymerizes DNA from a primer annealed to a DNA template in the presence of deoxyribonucleotide triphosphates. It has an optimum temperature of 75°C and can survive repeated incubations at > 95°C. Features and benefits High performance: highly purified thermostable DNA polymerase for PCR Reliable: highly purified recombinant enzyme, providing excellent batch-to-batch reproducibility and minimized nuclease contamination 20 Product Quantity Product code Sera-Mag select 5 mL 29343045 Sera-Mag select 60 mL 29343052 Sera-Mag select 450 mL 29343057 Case study Case study: Sera-Mag select in a targeted resequencing approach. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Sera-Mag select size selection and PCR clean-up reagent For use in NGS Sera-Mag select size selection and PCR clean-up reagent is based on well-known, solid phase, reversible immobilization technology for selective binding of DNA fragments for applications such as NGS and PCR clean-up. It combines the convenience of magnetic bead technology, using the exceptional binding characteristics of Sera-Mag carboxyl speedbeads with an optimized binding solution in a ready-to-use formulation. Features and benefits Reliability: High yield in the recovery of specific fragment sizes for optimal sequencing efficiency Simplicity: One product for size selection and PCR clean-up instead of two separate products Minimal disruption: Follows standard protocols for size selection and PCR clean-up, allowing for direct integration into existing workflows Next-generation sequencing (NGS) has lowered the cost and time required for sequencing, making genomic-level insights more accessible to academia and industry. Here are our 10 top tips to help you further reduce the cost of your NGS workflows, whether you only sequence occasionally or run a series of high-throughput platforms. 1: Start using NGS This might seem like an odd tip to start with, but Sanger sequencing is still in heavy use across academia and industry and many might not have considered trying NGS yet. Sanger sequencing has its purposes. If you need to check the integration of a short vector or gene editing insert, sequence a single gene, or perform some basic microsatellite analysis, Sanger sequencing is the way to go. For anything more substantial, such as sequencing of a whole chromosome or genome or identifying novel variants, NGS provides a far more comprehensive data output. Amplicon sequencing on small NGS platforms can be an ideal stepping stone where your throughput allows. NGS opens new possibilities for experimental design and, as outlined in a later tip, does not need you to have a lot of specialist knowledge to get started, and it can have a considerably lower cost-per-kilobase sequenced than Sanger sequencing. 2: Try third-party consumables for NGS Sequencing services and platform vendors are likely to recommend specific (often their own) kits for sample and library preparation. They might claim these kits are optimized for or validated on their 10 top tips to help reduce the cost of your NGS workflow $ 21 4: Amplify with Phi NGS workflows can be sample-hungry, fraught with variation, and create challenges when your sample is precious and in limited supply. In many cases, you should consider a sample amplification step as essential for doing the required analysis and also providing security in case repeats are required. The traditional approach, PCR, often requires optimization. Primers bind some areas of the genome more efficiently than others. Thermal cycling can introduce bias and vary in efficiency depending on the quality of the reagents and the thermal cycler you use. Phi29 polymerase provides reliable and isothermal amplification of circular and linear DNA using random primers. A short reaction at 30°C can amplify the picograms of DNA from a single cell to micrograms for sequencing, generating fragments often over 70 kb in size. This high-fidelity and highly processive polymerase removes a considerable amount of doubt in the amplification process, and minimizes the reliance on PCR to make the amplification step more reliable and efficient. 5: Automate your NGS workflow Automation is not just for large high-throughput sequencing facilities. The entry costs for automation are lower than they have ever been, and basic liquid handling systems need not take more space than a laminar flow hood. Automating these basic tasks both reduces your hands-on time, freeing up members of your team for other projects, improves the overall speed of the workflow, and reduces the chances of errors. 6: Track your samples automatically (or at least easily) Like automation, using barcodes to track and maintain a database of samples and their properties is no longer just for large facilities. The result of mixing up two samples or incorrectly labeling data is just as concerning for the academic researcher as it is for sequencing service providers. Hand-labeling samples and noting their specifics in a notebook might work well enough for a few tubes. But as batch and database sizes increase, there is a likelihood of introducing errors. These errors then lead to staff collecting, preparing, and sequencing samples again, an unnecessary waste of time and resources. Barcodes and readers are readily available and reduce the risks of cross-contamination and sample mix-ups. Many entry-level plate handling and storage systems also incorporate barcode readers, so a modestly-sized facility can also benefit from automatic sample tracking. platforms, however, the ubiquitous nature and understanding of NGS technologies now means third-party consumables can be both more cost-effective and perform just as well or better than vendor kits. Third-party kits provide you with flexibility for experimental design, and having the ability to shop around for the best performance, workflows, and deals means you can improve efficiency while keeping costs down. For example, rather than separate reagents for isolation, size selection, and PCR clean-up—key steps in sample and library preparation—you could use magnetic bead-based kits to have a single solution and automatable workflow. 3: Magnetize your sample (and library) preparation Speaking of magnetic beads, it is worth highlighting just how versatile these super(paramagnetic) little particles are in the NGS workflow. Magnetic beads are well suited to DNA extraction, size selection, and clean-up as alternatives to column-based purification, providing flexibility in scale and handling. This approach can greatly simplify extraction and size selection steps while introducing a level of consistency that is challenging to achieve otherwise between different staff and labs. Magnetic beads can also simplify library preparation for multiplexed sequencing and are readily automatable. Find out more about these in the tips below. 22 7: Scale down volumes to scale up numbers and cut costs It is common to work in the 96-well format, especially when handling samples manually. An 8-way pipette and a steady hand makes preparing the occasional plate of samples for sequencing quick and easy. However, if you tend to have larger batches of samples, and might already be considering automation, it could also be worth switching to a 384-well format. Not only will the smaller format reduce the number of plates and reagent necessary, but combine it with automation and you could process samples with less variation and waste. 8: Multiplexing need not be perplexing In Sanger sequencing, one run equals one short sequence. In NGS, indexing adapters or molecular barcodes mean one run can equal hundreds or even thousands of libraries of sequences. Multiplexing like this is what has driven down the cost of sequencing. It enables you to run multiple libraries on a single flow cell, making maximum use of the capacity of your NGS platform and so make it more cost-effective. Multiplexing is not without its challenges though, one of which is normalization of NGS libraries. DNA normalization is the process of equalizing the concentration of DNA libraries for multiplexing so that no library is over- or underrepresented in the sequencing output. Usually the process involves careful measurement and dilution of the libraries, but once again, magnetic beads can provide a remarkably simple option that actually does away with the quantitation step altogether. 9: Know when to scale up and scale down Increasing parallel sequencing capacity has been essential in reducing the costs of NGS. In addition to multiplexing, some sequencing platforms provide you with the option to run multiple flow cells, effectively multiplying your sequencing capacity without also multiplying your costs. On the other hand, when your needs are more modest, these large high-throughput sequencing platforms can seem like a substantial investment with capacity far beyond your current requirements. That is where small, benchtop sequencing platforms can make more sense. These sequences can integrate easily into your lab, and are well suited for whole genome sequencing of microbes and viruses, as well as targeted gene sequencing. 10. Outsource sequencing and storage Ultimately, though, it is worth asking your team whether they need in-house sequencing right now. Any sequencing platform is a longterm investment, and will involve upkeep and optimization. As an alternative, you could take advantage of the many sequencing service providers out there. They will have established and optimized workflows that turn your prepared samples into reliable data. You could also reduce the burden of bioinformatics by using readymade cloud apps to look after your projects and process your data. And what about all that data? You could potentially store terabytes of data locally, but an off-site backup is essential for security and redundancy. This might be in the form of cloud-based storage for recent or regularly-accessed data, or tape-based cold storage for archiving data. Those same sequencing providers might also offer dedicated NGS data storage options, which could be a cost-effective option as they will already have their own data safeguarding measures in place, saving you the effort. 23 24 Product Quantity Product code Sera-Mag carboxylate-modified [E7] magnetic particles 15 mL 24152105050250 Sera-Mag carboxylate-modified [E7] magnetic particles 100 mL 24152105050350 Sera-Mag carboxylate-modified [E7] magnetic particles 1000 mL 24152105050450 Sera-Mag carboxylate-modified [E3] magnetic particles 15 mL 44152105050250 Sera-Mag carboxylate-modified [E3] magnetic particles 100 mL 44152105050350 Sera-Mag carboxylate-modified [E3] magnetic particles 1000 mL 44152105050450 24 Explore Explore ways to optimize your magnetic bead workflow in ‘The scientist’s guide to magnetic beads’. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Sera-Mag carboxylate beads and speedbeads For use in Sanger sequencing and NGS Carboxylic groups on the surface of Sera-Mag speedbeads and Sera-Mag carboxylate-modified magnetic beads permit easy covalent coupling to target biomolecules of interest, such as proteins and nucleic acids, using convenient carbodiimide chemistry. The cauliflower-shaped surface, paired with proprietary Sera-Mag beads and speedbead chemistry, provides a large surface area and offers excellent sensitivity and low nonspecific binding for greater accuracy. This can maximize sample retention or reduce the amount of beads required. E3 and E7 refer to the different manufacturing process for the beads. E3 and E7 beads behave similarly, and we continue to provide both bead types to our customers for test and validation in their chosen application. Sera-Mag carboxylate beads Sera-Mag carboxylate-modified magnetic beads combine a fast magnetic response time and high binding capacity, sensitivity, stability, and physical integrity. Features and benefits Ease of use: Covalent coupling of proteins, nucleic acids, etc. to carboxyl groups on the surface using standard coupling technologies Convenient: Isolation, selection, and clean-up of nucleic acids or direct conjugation of specific oligos and enzymes 25 Request a sample Request a sample of magnetic beads. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Product Quantity Product code Sera-Mag speedbead carboxylate-modified [E7] magnetic particle 15 mL 45152105050250 Sera-Mag speedbead carboxylate-modified [E7] magnetic particle 100 mL 45152105050350 Sera-Mag speedbead carboxylate-modified [E7] magnetic particle 1000 mL 45152105050450 Sera-Mag speedbead carboxylate-modified [E3] magnetic particle 15 mL 65152105050250 Sera-Mag speedbead carboxylate-modified [E3] magnetic particle 100 mL 65152105050350 Sera-Mag speedbead carboxylate-modified [E3] magnetic particle 1000 mL 65152105050450 Sera-Mag speedbead carboxylate-modified magnetic particles Sera-Mag speedbeads have a second layer of magnetite applied through the same core shell design process. This second layer allows a reaction twice as fast as the Sera-Mag carboxylatemodified beads in the presence of a magnetic field. Speedbeads are especially useful when the reaction medium is highly viscous and in clinical assays requiring a faster magnetic response time. Features and benefits Convenient: Isolation, selection, and clean-up of nucleic acids or direct conjugation of specific oligos and enzymes Reliable: Fast, precise, and high binding capacity for sample preparation, nucleic acid isolation, proteomics, and immunoassay applications 26 Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Product Quantity Product code Sera-Mag streptavidin-coated magnetic particles – 2500 to 3500 (low) pmol per mg 1 mL 30152103011150 Sera-Mag streptavidin-coated magnetic particles – 2500 to 3500 (low) pmol per mg 5 mL 30152103010150 Sera-Mag streptavidin-coated magnetic particles – 2500 to 3500 (low) pmol per mg 100 mL 30152103010350 Sera-Mag streptavidin-coated magnetic particles – 3500 to 4500 (med) pmol per mg 1 mL 30152104011150 Sera-Mag streptavidin-coated magnetic particles – 3500 to 4500 (med) pmol per mg 5 mL 30152104010150 Sera-Mag streptavidin-coated magnetic particles – 3500 to 4500 (med) pmol per mg 100 mL 30152104010350 Sera-Mag streptavidin-coated magnetic particles – 4500 to 5500 (high) pmol per mg 1 mL 30152105011150 Sera-Mag streptavidin-coated magnetic particles – 4500 to 5500 (high) pmol per mg 5 mL 30152105010150 Sera-Mag streptavidin-coated magnetic particles – 4500 to 5500 (high) pmol per mg 100 mL 30152105010350 Sera-Mag streptavidin magnetic beads For use in NGS Sera-Mag streptavidin-coated magnetic particles Sera-Mag streptavidin-coated magnetic beads provide solid phase support in immunoassays and molecular biology applications. The magnetic streptavidin particles can also be used as a universal base particle for coating biotinylated proteins, oligos, or other ligands to the particle surface. Features and benefits Optimized application: Contains covalently bound streptavidin with low (2500 to 3500 pmol/mg), medium (3500 to 4500 pmol/ mg) or high (4500 to 5500 pmol/mg) biotin binding capacities, providing a choice of biotin-binding capacity High capacity and precision: For enrichment and targeted sequencing applications 27 Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Product Quantity Product code Sera-Mag speedbeads streptavidin-blocked magnetic particles 1 mL 21152104011150 Sera-Mag speedbeads streptavidin-blocked magnetic particles 5 mL 21152104010150 Sera-Mag speedbeads streptavidin-blocked magnetic particles 100 mL 21152104010350 Sera-Mag speedbeads streptavidin-blocked magnetic particles 1000 mL 21152104010450 Sera-Mag streptavidin-blocked magnetic beads Sera-Mag speedbeads streptavidin-blocked magnetic particles provide high biotin-binding capacity along with a strong affinity for targeted, biotin-labeled molecules such as nucleic acids, proteins, and peptides with very low nonspecific binding. When the beads are combined with any of these molecules, the strong noncovalent association between streptavidin and biotin ensures efficient capture of the target molecule. Features and benefits Increased throughput and precision: The speedbead particles combine fast reaction kinetics and low, nonspecific binding Optimized for demanding applications: Including bead-in PCR and enrichment 28 Magnetic separation racks Magnetic separation racks contain a neodymium magnet for small-scale protein purification and sample enrichment with magnetic beads. For sample volumes from 1.5 mL to 50 mL. Features and benefits Strong magnetic field, providing quick and efficient separation of magnetic beads from various liquid sample media used routinely for nucleic acid extraction procedures Easily remove the magnetic block from the plastic rack to allow rapid demagnetization of the samples Choose from three magnetic separation racks to meet your needs: magrack 6, magnetic separation rack 15 mL, and magrack maxi — for samples up to 1.5, 15, and 50 mL, respectively. Magnetic separation rack 15 mL Magnetic separation rack 15 mL is for sample volumes up to 15 mL. You can process up to six samples in parallel with easy pipette access. The magnetic field is strong, providing quick and efficient separation of magnetic beads from various liquid sample media, including viscous samples (e.g., blood or plasma diluted in typical viscid lysis/binding buffers used routinely for nucleic acid extraction procedures). The magnetic beads are quickly attracted to magnets located at the back of the magnetic separation rack. The bead pellet is distributed along just one side of the tube permitting maximum bead retention and yield. Easily remove the magnet block from the plastic rack to allow rapid demagnetization of the samples. Complementary to the following magnetic bead-based products: SeraSil-Mag silica coated superparamagnetic particles, Sera-Mag speedbeads carboxylate-modified magnetic particles, or Sera-Mag carboxylate-modified magnetic particles. Magrack 6 Magrack 6 is for sample volumes up to 1.5 mL. You can process up to six samples in parallel. Magrack 6 has an aluminum housing (blue) and detachable plastic bar (white) containing a neodymium magnet. The anodized aluminum housing assists with electrical conductivity. The neodymium magnet is extraordinarily strong, helping with magnetic reaction times as well as optimizing performance during protein purifications or enrichments. Complementary to the following magnetic bead-based products: protein A Mag Sepharose, protein A Mag Sepharose Xtra, protein G Mag Sepharose, protein G Mag Sepharose Xtra, His Mag Sepharose Ni, NHS Mag Sepharose, and Streptavidin Mag Sepharose. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 29 Product Product code Magnetic separation rack 15 mL 29710714 Magrack 6 28948964 Magrack maxi 28986441 Download If you’re unsure which magnetic beads to use for your application, check out our decision tree. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Magrack maxi Magrack maxi is for sample volumes up to 50 mL. It has an aluminum housing (blue) and a detachable plastic bar (white) containing a neodymium magnet. The anodized aluminum housing assists with electrical conductivity. The neodymium magnet is extraordinarily strong, helping with magnetic reaction times as well as optimizing performance during protein purifications or enrichments. Complementary to the following magnetic bead-based products: protein A Mag Sepharose, protein A Mag Sepharose Xtra, protein G Mag Sepharose, protein G Mag Sepharose Xtra, His Mag Sepharose Ni, NHS Mag Sepharose, and streptavidin Mag Sepharose. 30 Custom bead formats and types and conjugations and custom reagents Custom conjugation is possible for any of our protein and nucleic acid sample preparation, labeling, and detection products including Sera-Mag magnetic beads. Our dedicated customization experts will work with you every step of the way, from defining the product specifications to delivery completion. From completing complex conjugations and formulations to performing your quality control test before the beads and reagents leave our factory, we are equipped to meet your needs. Customization options include: Concentration: particles and reagents over a wide range of concentrations. Formulation: modifications to buffer composition, containers, dispense volumes. Conjugation: enzymes, antibodies, and custom ligands. Custom particles: nonstandard sizes or functionalities. Stabilization: lyophilization of the custom product — including magnetic beads — with our Lyo-Stable services. Packaging and kitting: from lab pack to bulk, and custom kits. We can provide historical batch data, batch-to-batch consistency, performance, buy specifications, technical insights, and studies to facilitate validation activities. Learn more Learn more about custom biology services. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 31 Detection Research and clinical diagnostic laboratories face different challenges and therefore have different needs when it comes to detection and sequencing technologies. Maybe simplicity is needed on a budget, perhaps more reliable results are sought from scarce samples, or possibly higher throughput is needed for maximum productivity. Whatever your need, choosing the most appropriate detection or sequencing platform is paramount. Detection for maximum productivity PCR is a relatively simple and widely used molecular biology technique to amplify and detect DNA and RNA sequences. A welloptimized reaction is therefore essential for accurate results. Quantitative PCR is used to detect, characterize, and quantify nucleic acids for numerous applications. RT-qPCR enables RNA transcripts to be quantified by reverse transcribing them into cDNA and then applying the qPCR process. It is inherently an analog process and quantitation relies on setting a threshold against predetermined standards. The power of qPCR across a diverse set of applications can be harnessed using algorithms, optimized master mixes, intuitive data analysis software, flexible instrumentation, and commercially available fluorescence-detecting thermocyclers to amplify specific nucleic acid sequences. With real-time PCR (RT-PCR), data is collected throughout the PCR process rather than at the end, which means that reactions are characterized by the point in time during cycling when amplification of a target is first detected rather than after a fixed number of cycles. Some of the benefits of RT-PCR include the ability to measure the initial concentration of target DNA over a range of five or six orders of magnitude, the lack of reliance on downstream analysis such as electrophoresis or densitometry, the possibility for multiple samples to be assessed simultaneously (which is ideal for high-throughput situations), its high sensitivity, and the provision of immediate information. A disadvantage is the equipment is more expensive than traditional PCR instruments. Manual and automated thermal cyclers and PCR products offer a direct route to reliable PCR results for researchers demanding efficiency and accuracy from sequencing to flow cytometry and real-time, digital, and end point PCR — from sample prep to data analysis. Sequencing is the process of reading the nucleotides present in DNA or RNA molecules. There are two types of sequencing technologies that are used today: Sanger sequencing and NGS. Each of these technologies has utility in today’s genetic analysis environment. Read more Read more in our knowledge article: ‘Best practices for PCR success’. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 32 Lyo-Stable™ lyophilization service Lyophilization, also known as freeze-drying or cryo-desiccation, is the process stabilizing a sample by removing water without applying heat. Besides increasing sample shelf life, freeze-drying samples also reduces sample weight and volume, reduces the risk of contamination, and eliminates the need to refrigerate samples, all of which decreases shipping and storage costs. Our patented lyophilization technology stabilizes individual proteins, reagents, and complete multiplex assays by providing a molecular environment that protects against conformational changes in protein structure. Your made-to-order cake or bead will contain our proprietary excipient mix to stabilize your temperaturesensitive assay for storage and shipping at ambient temperature. All you need to add is your sample, and you are ready to go! Contract manufacturing From simple buffers to multicomponent kits, large or small batch sizes, Cytiva can develop and manufacture custom products to simplify your workflows and supplement your capabilities. We can provide: Sourcing and validating raw materials Custom design services Custom formulations, volumes, and concentrations Custom packaging and labeling Custom testing and documentation Secured supply and delivery Scale-up capabilities to meet your needs Stability studies Full original equipment manufacturer (OEM) services Our analytical experience, combined with advanced instrumentation and stabilization technology, ensures precise control of the product as it progresses through manufacturing, resulting in a product that meets the highest standards and specifications. Batch-to-batch reproducibility is an important characteristic of each product we manufacture. We also offer full kitting and packaging services, including customer-designed packaging. Get in touch Get in touch to discuss your molecular diagnostics workflow. Learn more Learn more about our Lyo-Stable custom stabilization service. Learn more Learn more about our contract manufacturing services. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 33 Lab-based immunodiagnostics Sample collection Target capture Signal generation Detection Process Sample collected e.g. by blood draw, swab or collection of other body fluid into stabilization matrix to preserve content Sample is incubated with magnetic beads with antibody/antigen coating Antibody conjugate plus substrate are added to develop signal Developed signal is read in a suitable instrument that provides data output Cytiva products • Sera-MagTM carboxyl-modified magnetic beads and speedbeads • Sera-Mag streptavidin magnetic beads • Sera-Mag streptavidin-blocked magnetic beads • Sera-Mag amine-blocked magnetic beads Cytiva services Custom bead formats/types | Custom bead conjugations Lyo-StableTM lyophilization service | Contract manufacturing Click anywhere in the table to explore 34 Magnetic bead-based approaches in immunodiagnostics have become essential to meet the needs of healthcare today. While ELISAs are still popular, the benefits of magnetic beads — accuracy, speed, and simplicity — are driving us towards precision health. For immunoassays, magnetic particles such as Sera-Mag beads offer a convenient alternative to plate-based ELISAs and, as they rely on broadly the same assay principles, transition from plate to bead is straightforward in most cases. The same development considerations apply when building assays, with nonspecific binding and sensitivity being top of most people’s list of concerns. Having the assay on the bead provides a level of flexibility that cannot be achieved by ELISA, and manipulation in high-throughput settings becomes far easier. Enhancing ELISA with magnetic beads Enzyme-linked immunosorbent assay (ELISA) is a widely adopted biochemical technique in immunology and molecular biology. It plays a pivotal role in various applications, including diagnostics, biomedical research, and drug development. While the capabilities of ELISA are well-established, recent innovations using magnetic beads are increasing its usefulness. However, intricacies of ELISA including the integration of magnetic beads, can be challenging to grasp for beginners and even experienced researchers. Practical applications of ELISA ELISA is used in many different fields. In diagnostics, ELISA is extensively used in clinical laboratories for diagnosing diseases. It can detect specific antibodies or antigens associated with various diseases such as HIV, hepatitis, and autoimmune disorders. The technique can also be used to diagnose endocrine conditions such as thyroid disease and a variation of it is used in home pregnancy tests. ELISA is also used in nonmedical fields. For environmental monitoring, it is used to screen environmental samples for contaminants such as pollutants, allergens, and pathogens. It can also be used for checking food and beverage safety and quality control. Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection Sample collection Development of lab-based immunodiagnostics Almost all protein samples need further preparation after collection. The quality of such preparation can be critical to successful downstream analysis. So, giving sample preparation due attention should be the be the first consideration when developing any workflow. The phrase protein sample preparation can mean different things, and strict definitions are truly relevant only in a defined workflow. For example, the use of chromatography methods is often considered to be part of sample preparation if the purpose is protein fractionation in proteomics studies. But chromatography is less likely to be considered as part of sample preparation if you are isolating and/or detecting a single protein. Within immunodiagnostics, pharma development and other proteomics fields, there is increasing demand for high-quality, small-scale protein preparation for analytical purposes. As protein analysis has become complex and sensitive, the need for adequate screening techniques has likewise grown. 35 How magnetic bead-based ELISA works In a conventional ELISA, plates or wells coated with capture antibodies are used to immobilize target antigens from the sample. In magnetic bead-based ELISA, the capture antibodies are attached to magnetic beads instead. The process involves the following steps: Antigen binding: Magnetic beads, precoated with capture antibodies specific to the target antigen, are mixed with the sample. The magnetic beads bind to the antigen in the sample. Magnetic separation: A magnet is applied to the reaction mixture. The magnetic beads, along with the bound antigens are pulled to the side of the container, while unbound substances remain in solution. Washing and detection: The unbound substances are removed by carefully aspirating the supernatant. Then, detection antibodies, typically labeled with enzymes or fluorescent markers, are added. These detection antibodies bind specifically to the antigens on the magnetic beads. Signal amplification: As with traditional ELISA, an enzyme substrate is introduced, and a reaction occurs to produce a signal, which can be quantified. Magnetic bead-based ELISA has the advantage of signal amplification due to the close proximity of the magnetic beads, enhancing the assay’s sensitivity. Advantages of magnetic bead-based ELISA Using magnetic beads in your ELISA has many advantages. You’ll have enhanced sensitivity because magnetic beads allow for a higher binding capacity. This increased sensitivity is especially important when dealing with low-abundance antigens. Magnetic beads can also save the end user time. Assays are faster because magnetic bead-based ELISA often requires shorter incubation times. Magnetic beads are also automation-friendly, so assays using them are well-suited for high-throughput applications. These advantages could lead to faster and earlier diagnoses for patients allowing them to receive more timely treatment. Magnetic bead-based ELISA also has applications beyond diagnostics. Researchers use this technique in pharmaceutical research for drug discovery, pharmacokinetics, and pharmacodynamics studies. It is also valuable in protein-protein interaction studies and studying binding affinities. And genomic and proteomic researchers use magnetic bead-based ELISA for DNA and protein purification and analysis. The future of ELISA with magnetic beads Magnetic bead-based ELISA has been around for a while, but is likely to expand the use of ELISA in the following ways: Multiplexing: Magnetic bead-based ELISA allows for simultaneous detection of multiple analytes, making it well-suited for high-throughput applications. POC and portable devices: Compact and portable ELISA systems with magnetic bead integration could revolutionize POC testing, offering rapid diagnostics in remote or resource-limited settings. Customization for personalized medicine: Magnetic bead-based ELISA can be customized by selecting appropriate surface chemistries, coupling selected ligands to the beads, or using blocking agents. Designing in properties of the beads and/or sample processing and detection workflows can benefit personalized medicine, enabling tailored diagnostic and treatment approaches. Integration with emerging technologies: ELISA will continue to integrate with emerging technologies, such as microfluidics and nanomaterials, for improved sample handling and detection. Data integration and bioinformatics: Advanced data analysis and bioinformatics will play a crucial role in extracting meaningful insights from magnetic bead-based ELISA experiments. Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection 36 Conclusion ELISA is a valuable technique with a vast array of applications in research, diagnostics, and beyond. Understanding the principles and types of ELISA is key to obtaining reliable and meaningful results. The technique is evolving to meet the demands of modern research and healthcare. With an eye toward efficiency, sensitivity, and customization, automation, miniaturization, and magnetic bead-based ELISA represent a promising future, providing new avenues for innovation. Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection 37 Target capture Improving accuracy, speed, and simplicity in immunoassays Magnetic beads, or superparamagnetic particles, have been around for a while, and gained popularity in many applications, from antibody screening and immunoprecipitation (Mag Sepharose beads), to sample and library prep in NGS workflows (Sera-Mag magnetic beads). A key draw towards magnetic beads, in addition to the ease of use, is the array of surface chemistry options they offer, which provide as many functions as you can think of conjugations. Here are some examples of where immunodiagnostics is being used effectively in specific applications and what the future looks like. Immunodiagnostics in oncology and infectious disease Oncology and infectious disease cover a substantial portion of the market for immunodiagnostics. There are as many potential target biomarkers and combination of biomarkers as there are diseases. Molecular assays, NGS or PCR, can pick up known mutations linked to cancer. These hallmarks can help diagnose and even forecast disease development and response to different therapies. Immunodiagnostic assays can be designed to complement these molecular assays, providing a similar function for protein biomarkers. Immunodiagnostics complements molecular assays in the study of infectious diseases as well. While an infection is running its course, it might be relatively straightforward to use NGS or PCR to identify the pathogen. But, if the infection goes dormant, an immunoassay might be the only reliable way of identifying that pathogen. Why is this important? A dormant pathogen might reactivate later or react to what seems like a completely unrelated condition, therapy, or change in environment. The stipulations on treatment of multiple sclerosis with the monoclonal antibody Tysabri (natalizumab) makes a good example. If the patient has a dormant John Cunningham virus infection, they face an increased risk of developing progressive multifocal leukoencephalopathy (PML) when treated with this drug. So, an immunodiagnostic for antibodies against the John Cunningham virus is essential for any clinician considering Tysabri for their patient. Using immunodiagnostics in oncology In oncology, an immunodiagnostic test can confirm the presence of a solid tumor directly by detecting known tumor-associated antigens or indirectly by detecting antibodies against the antigens. Importantly, this approach might also flag the resurgence of a tumor or key changes in it that affect the efficacy of the therapeutic approach. Finding and developing assays for these biomarkers has become a significant trend in immunodiagnostics development, leading to complex screening tools and new technologies. For example, in early 2019, OPKO Health Inc. gained approval from the US Food and Drug Administration (FDA) for prostatespecific antigen (PSA) testing to improve the accuracy of prostate cancer diagnosis, ultimately with a view to reduce unnecessary prostate biopsies. OPKO Health’s test uses microfluidics, a recent development in immunodiagnostic approaches, to assay an array of biomarkers in multiplex. Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection 38 Product Quantity Product code Sera-Mag carboxylate-modified [E7] magnetic particles 15 mL 24152105050250 Sera-Mag carboxylate-modified [E7] magnetic particles 100 mL 24152105050350 Sera-Mag carboxylate-modified [E7] magnetic particles 1000 mL 24152105050450 Sera-Mag carboxylate-modified [E3] magnetic particles 15 mL 44152105050250 Sera-Mag carboxylate-modified [E3] magnetic particles 100 mL 44152105050350 Sera-Mag carboxylate-modified [E3] magnetic particles 1000 mL 44152105050450 Sera-Mag carboxylate beads Sera-Mag carboxylate-modified magnetic beads combine a fast magnetic response time and high binding capacity, sensitivity, stability, and physical integrity. Features and benefits Ease of use: Covalent coupling of proteins, nucleic acids, etc. to carboxyl groups on the surface using standard coupling technologies Convenient: Isolation, selection, and clean-up of nucleic acids or direct conjugation of specific oligos and enzymes Read more Read more in our knowledge article ‘Streamlining proteome analysis using SP3 — Single-pot, solid-phase-enhanced sample preparation (SP3) using magnetic beads is a simple and robust methodology for unbiased capture of total proteins’. Read more Read how the size of magnetic beads affects their properties and why you should care. Explore Explore ways to optimize your magnetic bead workflow in: ‘The scientist’s guide to magnetic beads’. Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection Sera-Mag carboxylate beads and speedbeads For use in chemiluminescence, colorimetric, turbidometric, ELISA, fluorescence Carboxylic groups on the surface of Sera-Mag speedbeads and SeraMag carboxylate-modified magnetic beads permit easy, covalent coupling to target biomolecules of interest, such as proteins and nucleic acids, using convenient carbodiimide chemistry. The cauliflower-shaped surface, paired with proprietary Sera-Mag beads and speedbead chemistry, provides a large surface area and offers excellent sensitivity and low nonspecific binding for greater accuracy. These 27ualities can maximize sample retention or reduce the amount of beads required. E3 and E7 refer to the different manufacturing process for the beads. E3 and E7 beads behave similarly, and we continue to provide both bead types to our customers for test and validation in their chosen application. 39 Product Quantity Product code Sera-Mag speedbead carboxylate-modified [E7] magnetic particle 15 mL 45152105050250 Sera-Mag speedbead carboxylate-modified [E7] magnetic particle 100 mL 45152105050350 Sera-Mag speedbead carboxylate-modified [E7] magnetic particle 1000 mL 45152105050450 Sera-Mag speedbead carboxylate-modified [E3] magnetic particle 15 mL 65152105050250 Sera-Mag speedbead carboxylate-modified [E3] magnetic particle 100 mL 65152105050350 Sera-Mag speedbead carboxylate-modified [E3] magnetic particle 1000 mL 65152105050450 Read more Read more about Reproducible protein and peptide cleanup for mass spectrometry using magnetic beads. Request a sample Request a sample of magnetic beads. Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection Sera-Mag speedbead carboxylate-modified magnetic particles Sera-Mag speedbeads have a second layer of magnetite applied through the same core shell design process. This second layer allows a reaction twice as fast as the Sera-Mag carboxylatemodified beads when in the presence of a magnetic field. Speedbeads are especially useful when the reaction medium is highly viscous, and in clinical assays requiring a faster magnetic response time. Features and benefits Convenient: Isolation, selection, and clean-up of nucleic acids or direct conjugation of specific oligos and enzymes Reliable: Fast, precise, and high-binding capacity for sample preparation, nucleic acid isolation, proteomics, and immunoassay applications 40 Product Quantity Product code Sera-Mag streptavidin-coated magnetic particles – 2500 to 3500 (low) pmol per mg 1 mL 30152103011150 Sera-Mag streptavidin-coated magnetic particles – 2500 to 3500 (low) pmol per mg 5 mL 30152103010150 Sera-Mag streptavidin-coated magnetic particles – 2500 to 3500 (low) pmol per mg 100 mL 30152103010350 Sera-Mag streptavidin-coated magnetic particles – 3500 to 4500 (med) pmol per mg 1 mL 30152104011150 Sera-Mag streptavidin-coated magnetic particles – 3500 to 4500 (med) pmol per mg 5 mL 30152104010150 Sera-Mag streptavidin-coated magnetic particles – 3500 to 4500 (med) pmol per mg 100 mL 30152104010350 Sera-Mag streptavidin-coated magnetic particles – 4500 to 5500 (high) pmol per mg 1 mL 30152105011150 Sera-Mag streptavidin-coated magnetic particles – 4500 to 5500 (high) pmol per mg 5 mL 30152105010150 Sera-Mag streptavidin-coated magnetic particles – 4500 to 5500 (high) pmol per mg 100 mL 30152105010350 Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection Sera-Mag streptavidin magnetic beads For use in chemiluminescence, colorimetric, turbidometric, ELISA, fluorescence Sera-Mag streptavidin-coated magnetic particles Sera-Mag streptavidin-coated magnetic beads provide solid phase support in immunoassays and molecular biology applications. The magnetic streptavidin particles can also be used as a universal base particle for coating biotinylated proteins, oligos, or other ligands to the particle surface. Features and benefits Optimized application: Contain covalently-bound streptavidin with low (2500 to 3500 pmol/mg), medium (3500 to 4500 pmol/ mg), or high (4500 to 5500 pmol/mg) biotin binding capacities, providing a choice of biotin-binding capacity High capacity and precision: For enrichment and targeted sequencing applications 41 Product Quantity Product code Sera-Mag speedbeads streptavidin-blocked magnetic particles 1 mL 21152104011150 Sera-Mag speedbeads streptavidin-blocked magnetic particles 5 mL 21152104010150 Sera-Mag speedbeads streptavidin-blocked magnetic particles 100 mL 21152104010350 Sera-Mag speedbeads streptavidin-blocked magnetic particles 1000 mL 21152104010450 Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection Sera-Mag streptavidin-blocked magnetic beads Sera-Mag speedbeads streptavidin-blocked magnetic particles provide high biotin-binding capacity along with a strong affinity for targeted, biotin-labelled molecules such as nucleic acids, proteins, and peptides with very low nonspecific binding. When the beads are combined with any of these molecules, the strong noncovalent association between streptavidin and biotin ensures efficient capture of the target molecule. Features and benefits Increased throughput and precision: The speedbead particles combine fast reaction kinetics and low, nonspecific binding Optimized for demanding applications: Including bead-in PCR and enrichment 42 Sera-Mag amine-blocked magnetic beads For use in chemiluminescence, colorimetric, turbidometric, ELISA, fluorescence Provides low nonspecific binding of proteins from the sample matrix. Sera-Mag speedbeads amine-blocked magnetic particles are uniform, colloidally stable, monodispersed, nonporous, superparamagnetic spheres made by a proprietary core-shell method. The core is a carboxylate-modified particle made by free radical emulsion polymerization of styrene and acid monomer. Magnetite (Fe3O4) is coated onto this core particle and then encapsulated with propriety polymers. The final surface is blocked using a proprietary method to help prevent nonspecific binding of proteins. Sera-Mag speedbeads amine-blocked particles combine fast magnetic reaction kinetics and high binding capacity due to their large surface area. Product Quantity Product code Sera-Mag speedbeads amineblocked particles 1 mL 19152104011150 Sera-Mag speedbeads amineblocked particles 5 mL 19152104010150 Sera-Mag speedbeads amineblocked particles 1000 mL 19152104010350 Explore Explore ways to optimize your magnetic bead workflow in ‘The scientist’s guide to magnetic beads’. Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection Features and benefits A nonsurfactant, nonprotein-blocked surface to reduce the undesired adsorption of proteins from a sample matrix. Fast reaction kinetics increase throughput and precision, also enabling faster movement through viscous solutions. Unique cauliflower-like surface provides increased area for binding reactions compared to smooth surface particles. Uniform, nominal 1 μm diameter provides excellent lot-to-lot reproducibility. 43 Magnetic separation racks Magnetic separation racks containing a neodymium magnet for small-scale protein purification and sample enrichment with magnetic beads. For sample volumes from 1.5 mL to 50 mL. Features and benefits Strong magnetic field, providing quick and efficient separation of magnetic beads from various liquid sample media used routinely for nucleic acid extraction procedures. Easily remove the magnetic block from the plastic rack to allow rapid demagnetization of the samples. Choose from three magnetic separation racks to meet your needs: magrack 6, magnetic separation rack 15 mL, and magrack maxi — for samples up to 1.5, 15, and 50 mL, respectively. Magnetic separation rack 15 mL Magnetic separation rack 15 mL is for sample volumes up to 15 mL. You can process up to six samples in parallel with easy pipette access. The magnetic field is strong, providing quick and efficient separation of magnetic beads from various liquid sample media, including viscous samples (e.g., blood or plasma diluted in typical viscid lysis/binding buffers used routinely for nucleic acid extraction procedures). The magnetic beads are quickly attracted to magnets located at the back of the magnetic separation rack. The bead pellet is distributed along just one side of the tube permitting maximum bead retention and yield. Easily remove the magnet block from the plastic rack to allow rapid demagnetization of the samples. Complementary to the following magnetic bead-based products: SeraSil-Mag silica coated superparamagnetic particles, Sera-Mag speedbeads carboxylate-modified magnetic particles, or Sera-Mag carboxylate-modified magnetic particles. Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection 44 Magrack 6 Magrack 6 is for sample volumes up to 1.5 mL. You can process up to six samples in parallel. Magrack 6 has an aluminum housing (blue) and detachable plastic bar (white) containing a neodymium magnet. The anodized aluminum housing assists with electrical conductivity. The neodymium magnet is extraordinarily strong, helping with magnetic reaction times as well as optimizing performance during protein purifications or enrichments. Complementary to the following magnetic bead-based products: protein A Mag Sepharose, protein A Mag Sepharose Xtra, protein G Mag Sepharose, protein G Mag Sepharose Xtra, His Mag Sepharose Ni, NHS Mag Sepharose, and Streptavidin Mag Sepharose. Product Product code Magnetic separation rack 15 mL 29710714 Magrack 6 28948964 Magrack maxi 28986441 Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection Magrack maxi Magrack maxi is for sample volumes up to 50 mL. It has an aluminum housing (blue) and a detachable plastic bar (white) containing a neodymium magnet. The anodized aluminum housing assists with electrical conductivity. The neodymium magnet is extraordinarily strong, helping with magnetic reaction times as well as optimizing performance during protein purifications or enrichments. Complementary to the following magnetic bead-based products: protein A Mag Sepharose magnetic particles, protein A Mag Sepharose Xtra, protein G Mag Sepharose, protein G Mag Sepharose Xtra, His Mag Sepharose Ni, NHS Mag Sepharose, and streptavidin Mag Sepharose. 45 Custom bead formats/types and conjugations Custom conjugation is possible for any of our protein and nucleic acid sample preparation, labeling, and detection products including Sera-Mag magnetic beads. Our dedicated customization experts will partner with you every step of the way, from defining the product specifications to delivery completion. From completing complex conjugations and formulations to performing your quality control test before the beads leave our factory, we are equipped to meet your needs. Concentration: particles over a wide range of concentrations. Formulation: modifications to buffer composition, containers, dispense volumes. Conjugation: enzymes, antibodies, and custom ligands. Custom particles: nonstandard sizes or functionalities. Stabilization: lyophilization of the custom product — including magnetic beads — with our Lyo-Stable services. Packaging and kitting: from lab pack to bulk, and custom kits. We can provide historical batch data, batch-to-batch consistency, performance, buy specifications, technical insights, and studies to facilitate validation activities. Get in touch Get in touch to discuss your immunodiagnostics workflow. Learn more Learn more about our custom Sera-Mag conjugation services. Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection 46 Signal generation The magnetic bead format has excellent properties for small-scale experiments. The high density of the beads allows rapid capture using magnetic devices. Samples are concentrated at the same time, which contributes to enhancing the signal in the analysis. In addition, the process is scalable to allow flexibility in volume of sample processed. Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection 47 Detection The earliest immunoassays were developed in the 1950s and relied on radiolabeling. This approach gave them sensitivity and accuracy. The technologies evolved rapidly, leading to the development of the ELISAs and enzyme immunoassays (EIAs) we use today for diagnostics. Both ELISAs and EIAs use an enzymatic reaction between, for example, horseradish peroxidase (HRP) and luminol to generate a color change that is proportional to the concentration of antigenbound antibody in the assay. Although they were two independently developed techniques, users often use the terms interchangeably. As well as ELISAs, we now have chemiluminescent immunoassays (CLIAs), florescent immunoassays (FIAs), and lateral flow assays (LFAs) in common use as in vitro diagnostic tests, each providing alternatives in methodology and reporting. The landscape is constantly changing, however. The big players in the market are pushing assay technology forward, adopting microfluidics and magnetic beads. Clinical scientists now have access to a dizzying array of immunodiagnostic products, with the list expanding all the time. Some of these immunodiagnostics target single biomarkers, but recent years have seen a trend towards detecting multiple analytes, demanded by the complexity and needs of making reliable early diagnoses. Whatever their targets and methodology, new immunodiagnostics need to encompass a few properties for market success: Sensitivity and accuracy with reproducible results Rapid workflow and results Simplicity, requiring minimal training Cost efficiency Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection 48 Lyo-Stable lyophilization service Lyophilization, also known as freeze-drying or cryo-desiccation, is the process of stabilizing a sample by removing water without applying heat. Besides increasing sample shelf life, freeze-drying samples also reduces sample weight and volume, reduces the risk of contamination, and eliminates the need to refrigerate samples, all of which decreases shipping and storage costs. Our patented lyophilization technology stabilizes individual proteins, reagents, and complete multiplex assays by providing a molecular environment that protects against conformational changes in protein structure. Your made-to-order cake or bead will contain our proprietary excipient mix to stabilize your temperature sensitive assay for storage and shipping at ambient temperature. All you need to add is your sample and you are ready to go! Contract manufacturing From simple buffers to multi-component kits, large or small batch sizes, Cytiva can develop and manufacture custom products to simplify your workflows and supplement your capabilities. We can provide: Sourcing and validating raw materials Custom design services Custom formulations, volumes, and concentrations Custom packaging and labeling Custom testing and documentation Secured supply and delivery Scale-up capabilities to meet your needs Stability studies Full OEM services Our analytical experience, combined with advanced instrumentation and stabilization technology, ensures precise control of the product as it progresses through manufacturing, resulting in a product that meets the highest standards and specifications. Batch-to-batch reproducibility is an important characteristic of each product we manufacture. We also offer full kitting and packaging services, including customer-designed packaging. Learn more Learn more about our Lyo-Stable custom stabilization service. Learn more Learn more about our contract manufacturing services. Process Cytiva products Cytiva services Sample collection Target capture Signal generation Detection 49 Sample collection Nucleic acid extraction/isolation Amplification and enrichment Detection Process Sample collected e.g. by blood draw, swab or collection of other body fluid into stabilization matrix to preserve content Sample is prepared and nucleic acid is isolated Specific sequences are amplified with primers/probes LAMP redout, quantative PCR (qPCR) Cytiva products • Bespoke sample preparation support can be provided on request • WhatmanTM NucleporeTM track-etched membranes • Whatman CycloporeTM track-etched membranes • Whatman AnoporeTM inorganic membranes • Whatman cellulose fiber pads • Whatman glass fiber pads • VividTM antigen clarification media • Various magnetic beads may also be an option here • Nucleotides • Taq DNA polymerase Cytiva services Customisation of membranes/ paper Custom reagents Lyo-Stable lyophilization services Point of care molecular diagnostics Click anywhere in the table to explore 50 Sample collection Sample is collected (e.g., by blood draw, swab, or collection of other body fluid) into a stabilization matrix to preserve content. Bespoke sample preparation support can be provided upon request. Development of assays for point-of-care molecular diagnostics Molecular testing in remote areas of the world is becoming a real possibility thanks to the developments in point-of-care testing devices that could be game changing for diagnosis of diseases such as tuberculosis, drug-resistant tuberculosis, HIV, and Ebola virus. POC devices often contain highly sensitive membranes to effectively capture the cells or microorganisms for further analysis. Bespoke sample preparation support can be provided upon request. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Key breakthroughs in molecular POC testing Adapting molecular diagnostics techniques for use in POC testing has been no easy feat, with the need to increase the speed, portability, and simplicity of lab-based techniques; all while retaining high sensitivity and specificity. A key breakthrough in POC testing has been the use of microfluidic technologies to create “lab-on-a-chip” POC assays. These assays complete all sample, analysis, and detection reactions within microfluidic channels. As a result, lab-on-a-chip assays are small, portable, and require very little sample and reagent. They drastically reduce costs and turnaround times and can enable the detection of multiple analytes. Several types of diagnostic techniques have successfully been used in molecular POC tests, including polymerase chain reaction (PCR), isothermal amplification techniques, and CRISPR. Faster, more portable PCR testing PCR, particularly reverse transcriptase-PCR (RT-PCR) and realtime PCR (qPCR) are the gold standard techniques in molecular diagnostics. These techniques can be extremely sensitive, with the ability to amplify and produce a detection signal from a single DNA or RNA molecule. Several RT-PCR systems that use microfluidic technology including the Cepheid GeneXpert have been granted the clinical laboratory improvements amendments (CLIA) waiver by the United States Food and Drug administration (FDA), allowing their use in POC settings. These systems typically use a single cassette that contains separate chambers for sample preparation, amplification, and detection. The small reaction volumes used in these microfluidic systems vastly reduce the time required for thermocycling, providing results within 30 minutes. So far, the GeneXpert system has been used in many tests, including for the multiplex detection of COVID-19, influenza A and B, and the multiplex detection of several human papillomavirus (HPV) oncotypes (1–3). As microfluidic technology and other test components advance, we should expect to see a further increase in the speed of PCR-based POC tests (Fig 1). Fig 1. PCR system designed with a microfluidic chip. 51 Rapid and simple POC testing using isothermal amplification Isothermal amplification methods are arguably better suited to POC testing than PCR, with low-cost, single-tube assays that don’t require thermocycling steps. Loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA) are the two most commonly used isothermal amplification techniques in molecular POC tests. LAMP-based devices In recent years, several LAMP systems with a reverse transcription step (RT-LAMP) have been increasingly used in POC settings. These systems are table-top platforms with kits available for detecting various infectious diseases. A user simply mixes a patient sample with kit reagents as directed, places tubes into the machine for amplification, and receives results in less than an hour. The tests can even be done without extracting nucleic acids from the sample. Diseases that can be diagnosed include malaria, strep A and strep B, HPV, whooping cough, and sexually transmitted infections within just 30 minutes (4, 5). The development of LAMP-on-a-chip systems is further improving the speed and accessibility of LAMP-based testing (Fig 2). These devices can detect multiple HPV types in just 15 minutes and have even been used to detect COVID-19 within just 3 minutes, being the fastest COVID-19 test reported as of 2023 (6, 7). Fig 2. The key steps and components for the Integration of LAMP with microfluidics. 52 Lateral flow strips integrating RPA RPA is another rapidly advancing molecular diagnostic technique used in POC testing (Fig 3). Its main advantage is the low isothermal amplification temperature of 37°C to 42°C, with amplification even possible at room temperature. Another key advantage of RPA is that the labeled amplification products can be easily detected using lateral-flow strips. So you get the benefits of lateral-flow tests, including their low-cost, simple components, and rapid development along with RPA’s high accuracy and fast turnaround times. RPA lateral-flow tests have been used to detect several infectious diseases such as COVID-19, Salmonella, the tropical disease Schistosoma mansoni, and the multiplex detection of Dengue virus serotypes (8–10). Combining isothermal amplification with CRISPR Despite the many positives of isothermal amplification-based POC tests, a major downside is their risk of nonspecific amplification, which could potentially lead to false positive results. To overcome this, scientists are developing POC tests that integrate isothermal amplification with the highly accurate gene editing technology, CRISPR. In these tests, DNA is first amplified using isothermal amplification and then detected using CRISPR/Cas systems. And it all happens in a single tube. A range of different CRISPR-isothermal POC tests have been developed including those to detect COVID-19, malaria, and African swine fever. These tests have shown high performance, with some even boasting 100% specificity and sensitivity (11–13). Fig 3. (A) Schematic Diagram: Illustrating the fusion of RPA with LFD, featuring FAM (carboxyfluorescein) marker. (B) Diagrammatic Sketch: Outlining the structure of LFD, with test and control lines. (C) Results: Depicting positive and negative outcomes. Protein in bulk solution Transport to interfacial region Rearrangement on surface Adsorption and surfactant interation NC fiber 53 54 Nucleic acid extraction and isolation What are track-etched membranes and why should you use them? Track-etched membranes (TEMs) are manufactured from highquality polycarbonate (PC) or polyethylene terephthalate (PET) film and are used to separate particles from solutions. Unlike traditional membranes, the holes of a TEM are all the same size. You can imagine a kitchen colander shrunk down until the holes are micron or even submicron size. Because the holes in a TEM are uniform, these membranes are often called true-pore membranes. What’s special about the pores in a TEM is that they have a very narrow size distribution. The size distribution of the pores in other membranes (sometimes called tortuous-path membranes) is a wide and flat curve, whereas the size distribution of the pores in TEMs forms a sharp peak. What’s also special is that the pore diameter can be controlled, so you can order a membrane that has pores that are the right size to capture a specific target while letting smaller contaminants filter through. You can also order membranes with a specific pore density to get the flow rate that you need. Track-etched membranes and diagnostics Track-etched membranes could be the next big thing in diagnostics — particularly for POC use. One of the goals in diagnostics is to get accurate results. You get more accurate results by having cleaner samples. And that’s where TEMs come in. They can be used to filter samples to isolate a target. The TEM makes the samples more uniform, so you get a better result. The membrane works very well as a prefilter for a diagnostic test because it allows the sample to be cleaned by the TEM before the purified sample is released into the detection step of the assay for analysis. As such, a TEM can be added to a lateral-flow test for POC or direct patient use to prefilter a sample before it is deposited on the sample pad. TEMs can be incorporated into different kinds of POC diagnostic tests. You could design a system in which a person loads a sample that goes through TEMs and comes out ready for PCR. Or better yet, have an automated system that does the PCR as well. This system could be standardized to allow you to create many different assays on the same automated system that can be used to diagnose different diseases. Incorporating TEMs into POC diagnostic assays can increase the speed and efficiency of the tests. You can quickly take a fresh sample and purify it with very little loss of the target molecule. TEMs will also reduce the amount of training needed for the people who are handling samples. Using the membranes can be almost as simple as a lateralflow test making them ideal for use in doctors’ offices or hospitals. Read more Read more about the manufacturing process and applications for track-etched membranes. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Download Track-etched membranes at work: Case studies in diagnostics. 55 Product Quantity Size Thickness Product code Nuclepore polycarbonate hydrophilic membranes 100 per pack 37 mm 0.6 μm 10417309 Nuclepore polycarbonate hydrophilic membranes 100 per pack 13 mm 5 μm 10417401 Nuclepore polycarbonate hydrophilic membranes 25 per pack 142 mm 0.2 μm 10417031 Nuclepore polycarbonate hydrophilic membranes 100 per pack 50 mm 5 μm 10417414 Nuclepore polycarbonate hydrophilic membranes 100 per pack 25 mm 0.6 μm 10417206 Nuclepore polycarbonate hydrophilic membranes 100 per pack 19 mm 0.8 μm 10417304 Nuclepore polycarbonate hydrophilic membranes 100 per pack 13 mm 0.8 μm 10417501 Nuclepore polycarbonate hydrophilic membranes 100 per pack 50 mm 0.4 μm 10417114 Nuclepore polycarbonate hydrophilic membranes 25 per pack 90 mm 0.4 μm 10417118 Nuclepore polycarbonate hydrophilic membranes 100 per pack 50 mm 0.2 μm 10417014 Nuclepore polycarbonate hydrophilic membranes 100 per pack 25 mm 5 μm 10417406 Table continued > Nuclepore™ track-etched membranes/papers Nuclepore™ track-etched hydrophilic membrane filters offer a highly defined pore size cutoff in the submicron range, enabling particle deformity measurement and capture of extremely small particles. These membranes come in a range of pores sizes and diameters and are also available in sheet format. Whatman Nuclepore membranes are suitable for a wide range of sample types and applications, including bioassays, cell biology, and environment analysis. Track-etched for highly defined pore size and high pore density Treated with polyvinylpyrrolidone (PVP) to render it hydrophilic Low extractables, minimizing filtration sample contamination risk High chemical resistance and good thermal stability for a wide range of sample types Low, consistent ash and low tare weights Smooth flat membrane surface for high particle visibility The sharply defined cylindrical micropores, combined with high flow rates and chemical and thermal resistance, make Nuclepore filtration membranes suitable for a variety of samples and applications, including microscopy, cytology, parasitology, and certain air and oceanography analyses. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 56 Product Quantity Size Thickness Product code Nuclepore polycarbonate hydrophilic membranes 25 per pack 210 × 293 mm 0.8 μm 10417551 Nuclepore polycarbonate hydrophilic membranes 25 per pack 8 × 10 in 0.2 μm 10417050 Nuclepore polycarbonate hydrophilic membranes 25 per pack 142 mm 12 μm 10418731 Nuclepore polycarbonate hydrophilic membranes 100 per pack 50 mm 2 μm 10418514 Nuclepore polycarbonate hydrophilic membranes 25 per pack 293 mm 0.4 μm 10417139 Nuclepore polycarbonate hydrophilic membranes 100 per pack 47 mm 3 μm 10418312 Nuclepore polycarbonate hydrophilic membranes 100 per pack 25 × 80 mm 5 μm 10417450 Nuclepore polycarbonate hydrophilic membranes 100 per pack 25 mm 1 μm 10418706 Nuclepore polycarbonate hydrophilic membranes 100 per pack 25 mm 10 μm 10418406 Nuclepore polycarbonate hydrophilic membranes 100 per pack 25 mm 3 μm 10418306 Nuclepore polycarbonate hydrophilic membranes 100 per pack 13 mm 1 μm 10418701 Nuclepore polycarbonate hydrophilic membranes 100 per pack 13 mm 3 μm 10418301 Nuclepore polycarbonate hydrophilic membranes 100 per pack 25 mm 12 μm 10418506 Nuclepore polycarbonate hydrophilic membranes 100 per pack 25 mm 15 μm 800203 Nuclepore polycarbonate hydrophilic membranes 100 per pack 293 mm 0.05 μm 112803 Get in touch Unsure of which membrane to use? Our team of experts are trained and can advise on which materials are most suitable for your application. Contact your local Diangostic Account Manager to discuss your needs. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Product Quantity Size Thickness Product code Nuclepore polycarbonate hydrophilic membranes 100 per pack 25 mm 0.8 μm 10417306 Nuclepore polycarbonate hydrophilic membranes 100 per pack 13 mm 0.2 μm 10417001 Nuclepore polycarbonate hydrophilic membranes 100 per pack 25 mm 2 μm 10418806 Nuclepore polycarbonate hydrophilic membranes 100 per pack 47 mm 10 μm 10418412 Nuclepore polycarbonate hydrophilic membranes 100 per pack 19 mm 1 μm 10418704 Nuclepore polycarbonate hydrophilic membranes 100 per pack 47 mm 1 μm 10418712 Nuclepore polycarbonate hydrophilic membranes 25 per pack 90 mm 1 μm 10418818 Nuclepore polycarbonate hydrophilic membranes 25 per pack 90 mm 2 μm 10418318 Nuclepore polycarbonate hydrophilic membranes 100 per pack 47 mm 3 μm 10418812 Nuclepore polycarbonate hydrophilic membranes 100 per pack 47 mm 12 μm 10418512 Nuclepore polycarbonate hydrophilic membranes 100 per pack 292 mm 0.2 μm 10417039 Nuclepore polycarbonate hydrophilic membranes 25 per pack 90 mm 1 μm 10418718 Nuclepore polycarbonate hydrophilic membranes 100 per pack 13 mm 10 μm 10418401 Nuclepore polycarbonate hydrophilic membranes 100 per pack 13 mm 12 μm 10418501 Nuclepore polycarbonate hydrophilic membranes 25 per pack 293 mm 1 μm 10418739 Nuclepore polycarbonate hydrophilic membranes 100 per pack 140 × 140 mm 0.6 μm 10417251 Nuclepore polycarbonate hydrophilic membranes 100 per pack 90 mm 12 μm 10418551 Nuclepore polycarbonate hydrophilic membranes 25 per pack 293 mm 0.2 μm 10417051 57 Product Diameter Pore size Product code Anodisc™ circle without support ring 13 mm 0.1 µm 6809-7013 Anodisc circle without support ring 13 mm 0.2 µm 6809-7023 Anodisc circle without support ring 47 mm 0.2 µm 6809-5522 Anodisc circle without support ring 13 mm 0.02 µm 6809-7003 Anodisc circle without support ring 47 mm 0.02 µm 6809-5502 Anodisc circle with support ring 25 mm 0.2 µm 6809-6022 Anodisc circle with support ring 25 mm 0.1 µm 6809-6012 Anodisc circle with support ring 47 mm 0.02 µm 6809-5002 Anodisc circle with support ring 47 mm 0.1 µm 6809-5012 Anodisc circle with support ring 25 mm 0.02 µm 6809-6002 Anodisc circle with support ring 47 mm 0.2 µm 6809-5022 Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Anopore™ inorganic membranes Anopore™ inorganic membranes are multifunctional and have a wide range of uses because of their innovative and precise pore characteristics. Aluminum oxide membranes have been used in an electrochemical detection system for the quantification of DNA, promising high sensitivity and specificity (1). They have also been used for the concentration and analysis of viruses (2). Anapore membranes have a precise, nondeformable honeycomb pore structure with no lateral crossover between individual pores. They are composed of a high-purity alumina matrix that is manufactured electrochemically. The membranes exhibit low protein binding, are nontoxic, and support cellular growth. Features and benefits High particle removal efficiency: Filters at precisely the stated cutoff, allowing no larger-sized particles to pass through the membrane. Particulate material is captured on the surface of the membrane for subsequent microscopic analysis. Transparency: When wet, the membrane is virtually transparent, so retained particles do not need to be transferred to another surface before microscopic examination. Compatibility: The membrane is hydrophilic and is compatible with most solvents and aqueous material. Minimal sample contamination: No monomers, plasticizers, adhesives, surfactants, or wetting agents are used in the manufacturing process, which removes sample contamination and ensures low protein binding and minimal loss of sample. 58 Cellulose fiber pads The base cellulose is a key part of the system, and the correct choice of absorbency, wicking rate, and wet strength are critical to producing a working assay. The Cytiva range of cellulose materials for dipstick colorimetric assays offers highly consistent and inert substrates for absorption of the active chemicals required for development of dipstick tests. CF1 CF1 is a light and thin, 100% cotton linter material (176 µm thickness at 53 kPA), suitable for use as a sample pad for small volume lateral-flow applications. Features and benefits Light, thin grade suitable for small volume. Consistent absorbency and wicking rates for test-to-test reproducibility. Product manufactured in controlled environments from highquality materials minimizes the risk of false results due to sample contamination. Low protein binding for high test sensitivity and minimal loss of analyte. Naturally hydrophilic for rapid rewetting after prolonged storage. Compatible with most housing styles. Minimal leakage along the strip to minimize the risk of contaminating test results. CF2 CF-2 is a wet-strengthened, cellulose fiber for use in dipstick colorimetric assays that contains an FDA-approved resin that binds the cellulose fibers together. Features and benefits High strength of the paper, especially when it is wet, reduces waste during manufacture due to the snapping of the cellulose during coating. Reduced risk of the pad decomposing during use. Easier handling during kit manufacture. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 59 CF3 CF3 is a medium weight, 100% cotton linter material. Consistent absorbency and wicking rates for test-to-test reproducibility. Product manufactured in controlled environments from highquality materials minimizes the risk of false results due to sample contamination. Low protein binding for high test sensitivity and minimal loss of analyte. Naturally hydrophilic for rapid rewetting after prolonged storage. Compatible with most housing styles. CF4 CF4 is a medium weight, 100% cotton linter material. Features and benefits Consistent absorbency and wicking rates for test-to-test reproducibility. Product manufactured in controlled environments from highquality materials minimizes the risk of false results due to sample contamination. Low protein binding for high test sensitivity and minimal loss of analyte. Naturally hydrophilic for rapid rewetting after prolonged storage. CF5 CF5 is a medium to thick weight, 100% cotton linter material. Features and benefits Consistent absorbency and wicking rates for test-to-test reproducibility. Product manufactured in controlled environments from highquality materials minimizes the risk of false results due to sample contamination. Low protein binding for high sensitivity and minimal loss of analyte. Naturally hydrophilic for rapid rewetting after prolonged storage. Compatible with most housing styles. Minimal leakage along the strip to minimizes the risk of contaminating test results. CF6 CF6 is a thick material suitable for high sample volume applications. Features and benefits Consistent absorbency and wicking rates for test-to-test reproducibility. Product manufactured in controlled environments from highquality materials minimizes the risk of false results due to sample contamination. Naturally hydrophilic for rapid rewetting after prolonged storage. Compatible with most housing styles. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 60 Grade Thickness (μm @ 53 kPA) Water absorption (mg/cm2) Dimensions Product code CF 1 176 18.7 22 mm × 50 m 8111-2250 CF 2 172 16.1 22 mm × 50 m 8112-2250 CF 3 322 34.6 22 mm × 50 m 8113-2250 CF 4 782 49.9 22 mm × 50 m 8114-2250 CF 4 782 49.9 210 × 293 mm 8114-6621 CF 5 954 63.3 22 mm × 50 m 8115-2250 CF 6 1450 136.3 22 mm × 50 m 8116-2250 CF 6 1450 136.3 20 mm × 50 m 8116-613 CF 6 1450 136.3 25 mm × 50 m 8116-7007 CF 6 1450 136.3 210 × 293 mm 8116-6621 CF 6 1450 136.3 25 mm × 50 m 8116-9915 CF 7 1873 252.3 22 mm × 50 m 8117-2250 Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection CF7 CF7 is a thick, 100% cotton linter, thick material, suitable for high sample volume applications. Features and benefits Consistent absorbency and wicking rates for test-to-test reproducibility. Product manufactured in controlled environments from highquality materials minimizes the risk of false results due to sample contamination. Low protein binding for high test sensitivity and minimal loss of analyte. Naturally hydrophilic for rapid rewetting after prolonged storage. Compatible with most housing styles. 61 Filter type/grade Abbreviation Grammage (g/m2) Thickness at 53 kPa (µM) Retention efficiency in liquid (µM) Whatman grade GF/B glass microfiber filter GF/B 143 675 1.0 Whatman grade GF/D glass microfiber filter GF/D 120 675 2.7 Whatman GF/DVA bound glass fiber filter GF/DVA 120 785 n/a Whatman grade GF/F glass microfiber filter GF/F 75 420 0.7 Whatman QM-A quartz microfiber filters QM-A 83 475 n/a Whatman GMF150 1 µm MG1(GD1) 145 730 1.0 Glass fiber pads for straightforward nucleic acid isolation Glass fiber filters are a popular and effective choice for nucleic acid extraction applications. Due to its high nucleic acid binding capacity, silica-containing materials, such as silica-coated superparamagnetic beads and glass fiber filters, are widely used as a binding material for nucleic acid extraction in commercial kits. But, with a range of glass fiber (GF) filters available, how do you choose the best grade for your application? Whether you’re a researcher, molecular diagnostic developer, or kit manufacturer, it is important to consider your final application when designing and optimizing a nucleic acid extraction method. Often, one of the key differences to consider is the size of the target DNA. So, does the grade of glass fiber filter really matter when it comes to nucleic acid extraction? And should the size of the target DNA influence your choice of glass fiber filter? We decided to investigate using a range of GF filters (Table 1). Table 1. Characteristics of Whatman glass fiber filters from Cytiva Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 62 Glass fiber filters are a popular and effective choice for nucleic acid extraction applications. So, does the grade of glass fiber filter really matter when it comes to nucleic acid extraction? And should the size of the target DNA influence your choice of glass fiber filter? We decided to investigate using a range of GF filters. We first explored how these different grades of glass filters performed for four sizes of DNA fragment: 3 Gb DNA from human blood 50 Mb DNA from tomato leaves 10 kb DNA plasmid 1.7 kb PCR product We standardized the tests using a commercially available DNA extraction kit, substituting the column and binding material for different glass filter grades (Fig 1). To serve as a comparison, we used the unaltered, commercially available column alongside our glass fiber filter columns, taking samples from the same lysate. Read more Read more about glass fiber filters for straightforward nucleic acid extraction. Choosing glass fiber filters for nucleic acid extraction Fig 1. Experimental setup: Nucleic acid-extraction of different DNA/RNA fragment sizesa. Sample Ready-to-use DNA Bind Lyse Wash Elute 63 MF1 MF1 is a bound glass fiber filter and is suitable for serum and whole blood samples around 100 μL Features and benefits Rapid: quick assays save valuable time with separations in 30 to 120 s Reproducible: no appreciable red cell hemolysis Reliable: our range of reliable materials adds flexibility in test design Flexible: choice of separation times allows for test optimization Convenient: separators appropriate for a range of blood volumes Optimized: extremely fast volume separation rate in relation to the volume of blood available LF1 LF1 is an untreated, bound glass fiber filter suitable for serum samples and even one drop of whole blood. Features and benefits Rapid: quick assays save valuable time with separations in 30 to 120 s Reproducible: no appreciable red cell hemolysis Reliable: our range of reliable materials adds flexibility in test design Flexible: choice of separation times allows for test optimization Convenient: separators appropriate for a range of blood volumes Optimized: fast volume separation rate in relation to the volume of blood available Grade Size Volume (per cm2) Thickness (μm @ 53 kPA) Wicking rate (s/4 cm) Water absorption (mg/cm2) Product code GF/DVA 22 m × 50 m > 50 μL 785 28.2 93 8145-2250 MF1 210 × 297 mm 10 to 50 μL 367 29.7 39.4 8122-6621 MF1 22 m × 50 m 10 to 50 μL 367 29.7 39.4 8122-2250 LF1 17 mm × 50 m 10 to 15 μL 247 35.6 25.3 8121-1750 Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Glass fiber pads Cytiva provides a broad portfolio of glass fiber filters in a range of formats and grades that can support nucleic acid extraction applications. GF/DVA GF/DVA is a bound glass fiber filter, especially suitable for saliva samples. Features and benefits Rapid: quick assays save valuable time with separations in 30 to 120 s Reproducible: no appreciable red cell hemolysis Reliable: our range of reliable materials adds flexibility in test design Flexible: choice of separation times allows for test optimization Convenient: Separators appropriate for a range of blood volumes Optimized: extremely fast volume separation rate in relation to the volume of blood available 64 Grade Pore size Average basis weight Product code Vivid™ ACG – antigen clarification glass fiber 13 mm 3.2 g/ft2 XE080CSH8X10 Read more Read more about antigen testing media. Request a sample Request samples of membranes and pads for your molecular assay development. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Vivid™ ACG — antigen clarification glass fiber This graded glass fiber material for POC sample clarification is comprised of a laminated glass fiber matrix with a highperformance acrylic binder attached to a polyester support. The material is specifically designed for sample clarification and is well-suited for preparation of sputum, nasopharyngeal, and saliva samples. The graded nature of this material allows for the processing of highly viscous samples and/or large sample volumes without compromising flow rates. The acrylic binder provides adequate strength for handling during use or in a manufacturing environment. Features and benefits Compatible with lateral-flow, cartridge PCR, traditional PCR design. Effective for the removal of contaminants from swab samples before applying to an in vitro diagnostic (IVD) test. Graded material allows for the processing of highly viscous samples and/or large sample volumes without compromising flow rates. Acrylic binder provides adequate strength for handling during use or in a manufacturing environment. Magnetic beads Various magnetic bead chemistries may be used for nucleic acid isolation. Please refer to the section on molecular assay development in the lab-based section. 65 Custom membranes The team has considerable experience and capabilities in the selection and customization of test components, as well as an understanding of the underlying science of rapid diagnostic test design. We can help with design robustness, work with different design formulations in parallel to minimize time and cost, and optimize test conditions so they are repeatable at both small- and large-scale quantities. Cytiva standard technology components for point-of-care immunodiagnostic assays can be customized to meet the requirements of your assay including slitting, cutting, punching, and a range of post treatment options. Our filtration specialists will work with you to develop the following components to your specific assay: Cellulose and glass fiber media Nitrocellulose membranes Capillary pore membranes such as track-etched polycarbonate membranes and Anopore inorganic membranes Whatman 903 Proteinsaver cards Our immunoassay development services provide early-stage developers access to our POC lateral-flow expertise, infrastructure, and proven development methodologies. Dr. Klaus Hochleitner, an industry-recognized expert in POC diagnostics development, leads our team of experienced specialists in state-of-the-art laboratories in Dassel, Germany. The team can help you solve challenges and optimize your prototype development, shortening time to commercialization. Learn more Learn more about our lateral flow immunoassay development services. Learn more Learn more about our custom immunoassay rapid test components. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 66 Product Quantity Product code Amersham dATP, solution, 100 mM 25 µmol 28406501 Amersham dATP, solution, 100 mM 100 mmol 28406502 Amersham dATP, solution, 100 mM 500 µmol 28406503 Amersham dATP, solution, 100 mM 100 µmol 28406512 Amersham dATP, solution, 100 mM 100 µmol 28406522 Amersham dATP, solution, 100 mM 25 mmol 28406531 Amersham dATP, solution, 100 mM 100 µmol 28406532 Amersham dATP, solution, 100 mM 25 µmol 28406541 Amersham dATP, solution, 100 mM 100 µmol 28406542 Amersham dNTP set (100 mM each A, C, G, T) 4 × 25 µmol 28406551 Amersham dNTP set (100 mM each A, C, G, T) 4 × 100 µmol 28406552 Amersham dNTP set (100 mM each A, C, G, T) 4 × 500 µmol 28406553 Amersham dNTP set (100 mM each A, C, G, T) 10 mmol 28406557 Amersham dNTP set (100 mM each A, C, G, T) 4 × 10 µmol 28406558 Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Amplification and enrichment Nucleotides Amersham dNTPs are high purity deoxynucleotides for amplification, dideoxy sequencing, labeling, mutagenesis, cDNA synthesis, and expression profiling. They are free from DNase, RNase, and nicking enzyme activity. Features and benefits Performance: Greater than 99% triphosphate purity (by HPLC) for consistency Convenient: Buffer-free, ready-to-use solutions at a variety of concentrations Tested: Functionally tested to produce a 20.7 kb PCR amplification product from λ-DNA Also available: high purity rNTPs providing easy-to-use solutions that save time for in vitro transcription. All rNTPs are lot tested for ribonuclease contamination. 67 Product Quantity Product code Taq DNA Polymerase (cloned) 250 27079804 Taq DNA Polymerase (cloned) 4 × 250 27079805 Taq DNA Polymerase (cloned) 10 × 250 27079806 Taq DNA Polymerase (cloned) 25 000 28937348 Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection Taq DNA polymerase Cloned Taq DNA polymerase is the recombinant protein form of the native enzyme from Thermus aquaticus expressed in E. coli. Like native Taq, it polymerizes DNA from a primer annealed to a DNA template in the presence of deoxyribonucleotide triphosphates. It has an optimum temperature of 75°C and can survive repeated incubations at > 95°C. Features and benefits High performance: highly purified thermostable DNA polymerase for PCR Reliable: highly purified recombinant enzyme, providing excellent batch-to-batch reproducibility and minimized nuclease contamination 68 Custom reagents Our dedicated customization experts will work with you every step of the way, from defining the product specifications to delivery completion. From completing formulations to performing your quality control test before the reagents leave our factory, we are equipped to meet your needs. Concentration: reagents over a wide range of concentrations Formulation: modifications to buffer composition, containers, dispense volumes Conjugation: enzymes, antibodies, and custom ligands Stabilization: lyophilization of the custom product with our Lyo-Stable services Packaging and kitting: from lab pack to bulk and custom kits. We can provide historical batch data, batch-to-batch consistency, performance, buy specifications, technical insights, and studies to facilitate validation activities. Learn more Learn more about our custom biology services. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 69 Detection What does the future hold for PCR? Quantitative polymerase chain reaction (qPCR) is a variation on the original PCR method developed in the 1980s that uses fluorescent probes to detect amplicons as they amplify. In diagnostics, qPCR assays can help determine the presence of genetic markers in real time, and therefore aid disease diagnosis. Quantitative PCR is inherently analog (i.e., the output is continuous), and quantitation relies on setting a threshold against standards. Digital PCR (dPCR) is a variation on qPCR that dilutes a single sample into many smaller reactions. Each reaction contains no more than one template molecule. Therefore, amplification can give you a discrete and absolute count of how many template molecules were present in the original sample and, importantly, a definitive answer on the presence of a marker. Should you use qPCR or RT-qPCR? If you’re interested in profiling gene expression rather than the gene itself, a variation of qPCR — reverse transcriptase qPCR (RTqPCR) — enables you to analyze the relative abundance of RNA. The reverse transcriptase converts the RNA to cDNA, which you can then treat in essentially the same way as a DNA sample. Pros and cons of qPCR and RT-qPCR Since the 1990s, qPCR has remained the most popular method in molecular diagnostics, despite the development of more modern approaches. Its continued success is largely the result of its four key benefits: sensitivity, simplicity, speed, and cost. Compared to other diagnostic assays, qPCR is a relatively cheap way to study genomic targets, even at tiny concentrations, with several reagents common to standard PCR such as the polymerase and nucleotides. A typical protocol is also fast — often providing same-day results — and the data is easy to interpret. A key drawback when choosing qPCR is that it isn’t really designed for interrogating complex or multigene conditions. You can only use so many probes in one assay, and so investigating many targets substantially increases PCR workload. This increased workload then affects costs and turnaround times. Another downside to PCR is the potential for biased outcomes. The approach requires preselection of targets, so the outcome of any assay is limited to the probe selection for that assay. Co-infections, for example, are easy to overlook with primers designed to pick up one infection and not another. Clinical applications for RT-qPCR Clinicians frequently turn to qPCR or RT-qPCR to identify pathogens. Together, they make up more than 50% of the diagnostic testing area. Infectious disease diagnosis (i.e., the identification of bacteria or viruses) can involve both DNA and RNA analyses. Infectious disease diagnosis often requires fast results, making RT-qPCR well suited. In human genetics, key areas for PCR are oncology and reproductive health. Clinicians can use PCR to detect, for example, minimal residual disease in leukemia patients, either through DNA or RNA. In reproductive health, PCR can pick up chromosomal aberrations at the preimplantation (in in vitro fertilization), prenatal, or postnatal stage of development. The future of clinical qPCR The overall trend in clinical DNA analysis is one of increasing scale. This trend is driven mainly by ever-decreasing costs of DNA sequencing. As a result, NGS-based assays are likely to replace PCR in applications where information from large or multiple regions of the genome determines diagnosis. Still, qPCR and RT-qPCR retain a large market share in molecular diagnostics and are likely to remain key players for the foreseeable future, particularly in applications where rapid results are critical. This need is most clear in the infectious disease market. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 70 Lyo-Stable lyophilization service Lyophilization, also known as freeze-drying or cryo-desiccation, is the process stabilizing a sample by removing water without applying heat. Besides increasing sample shelf life, freeze-drying samples also reduces sample weight and volume, reduces the risk of contamination, and eliminates the need to refrigerate samples, all of which decreases shipping and storage costs. Our patented lyophilization technology stabilizes individual proteins, reagents, and complete multiplex assays by providing a molecular environment that protects against conformational changes in protein structure. Your made-to-order cake or bead will contain our proprietary excipient mix to stabilize your temperaturesensitive assay for storage and shipping at ambient temperature. All you need to add is your sample, and you are ready to go! Learn more Learn more about our Lyo-Stable custom stabilization service. Process Cytiva products Cytiva services Sample collection Nucleic acid extraction / isolation Amplification and enrichment Detection 71 Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Process There is no general process. The analytes can be anything from small toxins over large proteins to nucleic acids. Select binding reagents and determine quantitative interaction kinetics from wither hybridoma culture supernatants (antibodies) or from in vitro synthesis (aptamers, affirmers). Other proteins as e.g. his-tagged molecule. Analytical column chromatography to develop purification protocols and to obtain small amounts of reagents for further tests followed by upscalling Determine binding kinetics for selected biomolecules in the prescence of membrane surfactants. Check for sufactants/ reagent combinations that show the smallest change of kinetic parameters compared to the original selection data Sample is collected (this can be all body fluids) and may need to be transfered to liquid media to allow application Specific antibody conjugates are captured at the test/control lines, typically showing up visually for detection Specific antibody conjugates are captured at the test/ control lines, typically showing up visually for detection Cytiva products • VIA Extractor tissue disaggregator • Cold enzymes mix for various tissue types • Biacore 8 series (= Biacore 8K and Biacore 8K+) • Biacore 1 series (= Biacore 1K, Biacore 1K+ and Biacore 1S+) • Series S sensor chip CM5 • Series S sensor chip CM3 • Series S sensor chip protein A (protein G and protein L) • Series S sensor chip SA • Series S sensor chip NTA • His capture kit type 2 • Biotin CAPture kit Series S • GST capture kit • Human antibody capture kit type 2 • Human Fab capture kit type 2 • Mouse antibody capture kit type 2 • Protein A Mag Sepharose • Protein G Mag Sepharose • Streptavidin Mag Sepharose • His Mag Sepharose • His Mag Sepharose Ni • His Mag Sepharose Excel • NHS Mag Sepharose • Affinity chromatography – HisTrap, GSTrap, MabSelect • Ion exchange chromatography – capto, RESOURCE • Gel filtration – Superdex, Superose • Biacore 8 series (= Biacore 8K and Biacore 8K+) • Biacore 1 series (= Biacore 1K, Biacore 1K+ and Biacore 1S+) • Series S sensor chip CM5 • Series S sensor chip CM3 • Series S sensor chip protein A (protein G and protein L) • Series S sensor chip SA • Series S sensor chip NTA • His capture kit type 2 • Biotin CAPture kit Series S • GST capture kit • Human antibody capture kit type 2 • Human Fab capture kit type 2 • Mouse antibody capture kit type 2 • Bespoke sample preparation support can be provided upon request • Whatman cellulose fiber pads • Whatman glass fiber pads • Whatman plasma separation pads • Whatman nitrocellulose membranes • Whatman nylon membranes • Whatman PVDF membranes • Whatman nitrocellulose membranes • Leukosorb media • Vivid plasma separation membranes Cytiva services Reagent characterization | Conjugate development | Full test development | Training and workshops Immunoassays for point-of-care use Click anywhere in the table to explore 72 Biomarker identification There is no general process. The analytes can be anything from small toxins over large proteins to nucleic acids. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 73 VIA Extractor™ tissue disaggregator Manual tissue dissociation is long, laborious, and affects cell viability. It is vital to choose equipment to speed up the single cell workflow that is simultaneously gentle on the cells and preserve the original cell state as much as possible. The VIA Extractor™ tissue disaggregator* disaggregates solid tissue into viable single cells. For use in high-throughput omics and flow cytometry research, the closed system uses mild processing to optimize yield and preserve content relative to the parent sample. Features and benefits Mild processing optimizes cell viability and yield and preserves content relative to the parent sample. Standardized, automated process reduces sample-to-sample variation. Disaggregates solid tissue into single cell suspension in as little as ten minutes. Adjust the processing speed, temperature, and time settings using the VIA Freeze™ Uno controlled-rate freezer to optimize results dependent on sample type and size. Process up to three samples in parallel in single-use pouches, minimizing the risk of contamination. Product Quantity Product code Omics applicator 60 29509359 Omics bundle (includes VIA Extractor tissue disaggregator*, the VIA Freeze Uno controlled-rate freezer and the Omics clamp) 1 29517120 Omics clamp 1 29509355 Omics pouch 10 29726921 *For research use only (RUO). Not for diagnostic use. Read more Read more about quality tissue dissociation for single-nuclei RNASeq preparation. Read more Read a comparative study of VIA Extractor tissue disaggregator for tissue dissociation for single-cell RNA sequencing analysis. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Please note that the VIA Extractor tissue disaggregator needs to be placed into the VIA Freeze Uno controlled-rate freezer to operate. If you are not in possession of a VIA Freeze Uno controlled-rate freezer, the Omics bundle should be ordered. The Omics bundle includes the VIA Extractor tissue disaggregator, the VIA Freeze Uno controlled-rate freezer and the Omics clamp. 74 Cold enzymes mix — kidney Effective enzymatic and mechanical dissociation of kidney tissue is necessary to complete single-cell analysis so that targeted treatments can be developed. Single-cell preparations of the renal tissue are quite cumbersome because the kidney is a complex organ, both functionally and anatomically. Each segment of the kidney is built by very distinct and specialized cell populations. Cold-temperature tissue dissociation is popular because it is thought to minimize cell death and, by consequence, improve the transcriptional profile of the single cells. The cold enzyme dissociation mix — kidney* is a standalone kit, intended for the dissociation of kidney tissue in singlecell sequencing. The kit has been optimized for use with semiautomated tissue disaggregation systems. Features and benefits Works effectively at 4°C: Minimized cell death and stress (defined as: apoptosis pathway markers, macrophages, etc.) and improved transcriptional profile of the single cells for more accurate results in single-cell sequencing and analysis. Ready-made solution specifically for kidney tissue: Optimized for the dissociation of the different segments that are operative in kidney tissue. Optimized for use with the VIA Extractor tissue disaggregator: One of the few systems that can sustain a cold temperature in a controlled manner. Effective dissociation at cooler temperatures increases viability and minimizes cell aggregation. Yields viable kidney cells: Single-cell sequencing provides the key to unlocking the origin of renal pathologies at the cellular level, including renal (kidney) carcinomas. One of the first and most critical steps in a single-cell RNA sequencing (scRNA-seq) protocol is the dissociation of tissues to yield fully-dissociated but intact and viable cells. Product Quantity Product code Cold dissociation enzyme mix – kidney* 1 pack × 20 reactions 29733434 *For research use only. Read more Read more about single-cell RNA sequencing and cold dissociation. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 75 Features and benefits Superb data quality while increasing your throughput eight-fold, compared to one-needle SPR systems Interaction analysis for screening, kinetics, affinity, epitope binning, concentration, and relative potency Higher operational efficiency with capacity options and streamlined assay development in parallel Modular configuration with option for analysis in a GxP regulated environment Rapid optimization of assay conditions and troubleshooting Product Quantity Product code Biacore 8K instrument kit 1 29722782 Biacore 8K+ instrument 1 29283382 Biacore 8K+ instrument and kit 1 29722783 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Protein selection Biacore™ 8 series (Biacore 8K and Biacore 8K+) Biacore™ 8 series efficiently delivers quality binding data to meet your toughest challenges in screening, characterization, process optimization, and quality control. Our 16 flow-cell surface plasmon resonance (SPR) systems, Biacore 8 series, come in two configurations: Biacore 8K for high-throughput screening and characterization Biacore 8K+ for a significantly higher capacity These eight-needle, high-sensitivity SPR systems rapidly provide reproducible kinetics, affinity and concentration data, and shortens your time to results by up to eight times compared to one needle systems. The blend of system flexibility and throughput reduces the experimental cycle time, even for complex targets and drug formats such as bispecific antibodies. This reduced cycle time offers more opportunity for screening in drug discovery and development of small molecules to large viruses in pure or complex samples. Application methods are easily transferred to other labs or other Biacore 8 series systems and to our one-needle SPR platform Biacore 1 series. 76 Biacore 1 series (Biacore 1K, Biacore 1K+, Biacore 1S+) Biacore 1 series is a one-needle platform that transforms protein interaction analysis using SPR — making it simpler and faster to use without compromising quality. Now you can focus on generating the consistent and reproducible data needed in your research. Our six flow-cell SPR system, Biacore 1 series comes in three system configurations: Biacore 1K, Biacore 1K+, and Biacore 1S+. The systems offer analytical flexibility that can grow with your group’s need for sample capacity, sensitivity, and throughput. All systems are built for analysis in a GxP-regulated environment. The single software platform for Biacore 1 series systems allow you to spend less time training on running the instruments and less time on result generation and evaluation. Application methods are easily transferred to other labs or to other Biacore 1 series systems and the higher-throughput systems of the Biacore 8 series. You can use Biacore 1 series systems across a wide range of applications, molecules, and both pure and complex samples — from small fragments to large viruses. The systems are scalable, so your research won’t be limited as your needs evolve. Features and benefits No programming skills needed for setup and to start analysis when using predefined methods and flexible software tools to speed up assay development Straightforward transfer of methods to other Biacore 1 series or Biacore 8 series systems Less time needed to train on running the instrument and how to generate and evaluate results Simpler data interpretation: compile, visualizev and export data with results in minutes Optimized injection design with six flow cells enables more efficient sample utilization and greater application utility, including multicomplex analyses in one run Maximize run efficiency by queuing methods and assays and let it run overnight or over the weekend Product Quantity Product code Biacore 1K SPR system 1 29726017 Biacore 1K+ SPR system 1 29726018 Biacore 1S+ SPR system 1 29726019 Read more Read our Biacore systems selection guide. Read more Read our Biacore consumables selection guide. Download Download our poster to save time and effort for your Biacore SPR interaction analysis study. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 77 Series S sensor chip CM5 Biacore sensor chip CM5 supports a range of immobilization chemistries to capture nucleic acids, peptides, antibodies, proteins, carbohydrates, or chemical compounds for interaction analysis from research to discovery, development, and quality control. The CM5 sensor chip carries a matrix of carboxymethylated dextran covalently attached to a gold surface. Molecules can be covalently coupled to the sensor surface by exploiting available amine, thiol, aldehyde, or carboxyl functional groups on the ligand. The CM5 dextran matrix extends about 100 nm from the gold surface and is flexible to allow relatively free movement of attached ligands. Sensor chip CM5 is the most versatile and widely used dextranbased chip, suitable for most general-purpose surface plasmon resonance (SPR) applications. Carboxymethyl surface chemistry supports the study of many different types of ligands and binding partners from small organic molecules, such as drug candidates, through to large molecular assemblies and even whole viruses. High binding capacity gives a high response, which is advantageous for capture assays and for interactions involving small molecules. High surface stability provides accuracy and precision and allows repeated analysis on the same surface. Application examples include: FBDD-based screening for novel drugs against Hepatitis C virus (HCV) Surface plasmon resonance-based determination of adenovirus concentration on Biacore T200 Features and benefits First choice for immobilization via -NH2, -SH, -CHO, -OH, or -COOH groups Attach proteins, antibodies, nucleic acids, carbohydrates, and small molecules (< 1000 Da) Compatible with a wide range of applications from basic research to quality control Suitable for ligand fishing and fragment-based drug discovery (FBDD) High immobilization capacity permits a broad range of capture densities Obtain real-time data on binding specificity, affinity, kinetics, concentration and more Product Quantity Product code Series S sensor chip CM5 Pack of 10 29149603 Series S sensor chip CM5 Pack of 3 BR100530 Series S sensor chip CM5 Pack of 1 29104988 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 78 Product Quantity Product code Series S sensor chip CM3 Pack of 1 29104990 Series S sensor chip CM3 Pack of 3 BR100536 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Series S sensor chip CM3 Biacore sensor chip with similar properties to sensor chip CM5, suited to large interaction partners and exploratory assay conditions. The shorter dextran chains of the CM3 allows the interaction to take place closer to the surface, which can improve sensitivity when working with large molecules, molecular complexes, viruses, or whole cells. Features and benefits Shorter dextran matrix and similar charge density to explore alternative assay conditions Use when the interaction partner in solution is very large Improves sensitivity when monitoring certain interactions Maintains sensitivity by keeping large particles as close as possible to the surface Attach proteins, nucleic acids, carbohydrates, or small molecules Couple to carboxyl groups on the sensor surface via -NH2, -SH, -CHO, -OH or -COOH. 79 Series S sensor chip protein A Biacore sensor chip for orientation-specific binding of antibodies (predominantly human) through Fc-region. The surface is a carboxymethylated dextran matrix with a recombinant protein A variant covalently attached. The surface ligand is the same as is used for our MabSelect SuRe™ protein purification products. Binds antibodies from several mammalian species, most notably human antibodies of the subclasses IgG1, IgG2, and IgG4 Binds only to the heavy chain within the Fc region, ensuring a specific orientation of the antibody on the surface Does not bind Fab fragments (unlike native protein A) Sensor chip protein A is a sensor surface pre-immobilized with a recombinant protein A variant binding antibodies for protein interaction analysis using Biacore systems. It is ready-to-use with high binding capacity offering a wide dynamic range and high convenience, reproducibility, and robustness. Sensor chip protein A is an excellent surface choice for antibody quantitation and characterization in biopharmaceutical, late-stage development and manufacturing QC. Product Quantity Product code Series S sensor chip protein A Pack of 3 29127556 Series S sensor chip protein A Pack of 1 29127555 Process Cytiva products Cytiva services Biomarker identification Protein selection Nucleic acid extraction / isolation Reagent purification Membrane family selection Sample collection Chromatography Detection Features and benefits MabSelect SuRe ligand on the surface Site-directed capture of antibody: binds predominantly to the heavy chain within the Fc region and ensures antibodies are bound to the surface in a specific orientation Time and effort savings: ready-to-use sensor chip eliminates the need to develop immobilization and regeneration conditions Confidence in data: highly controlled development processes and manufacturing conditions ensure reproducible results 80 Series S sensor chip protein G A Biacore Extend product used for oriented capture or binding of a wide range of mammalian and all human antibody subclasses. The surface is a carboxymethylated dextran matrix pre-immobilized with a recombinant protein G — GammaBind™ G, type 2. The recombinant protein G binds a broad range of IgG, such as human (including IgG3), rat, rabbit, mouse, guinea pig, goat, sheep, and cow. Sensor chip protein G is one of an expanding range of Biacore Extend products. These sensor chips provide researchers working in nonregulated environments with the convenience of a wider choice of consumables optimized for Biacore applications. What to expect from Biacore Extend product: Ordered as standard products Same customer support system is used as for a regular product QC performed, but no Certificate of Analysis/Conformance issued For some products, lot-to-lot variation might be higher compared to a standard product Features and benefits Combines convenience with high reproducibility and robustness in the analysis of interactions where antibodies are used as ligands Binds only to the heavy chain within the Fc region ensuring antibodies are bound to the surface in a specific orientation Ready-to-use, eliminating the need to develop immobilization and regeneration conditions, saving time and effort Product Quantity Product code Series S sensor chip protein G Pack of 1 29179315 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 81 Series S sensor chip protein L A Biacore Extend sensor chip for site-directed antibody capture. Suitable for study of a wide range of antibody fragments including Fabs and dAbs. The surface is a carboxymethylated dextran matrix pre-immobilized with a recombinant protein L for antibodies and antibody fragments containing kappa light chain subtypes (1, 3, and 4) without interfering with its antigen-binding site. Protein L is suitable for the capture of a wider range of antibody classes than both protein A and protein G, including IgG, IgM, IgA, IgE, and IgD. Protein L does not bind bovine immunoglobulins, which often contaminate serum supplements, and is therefore especially useful for quantifying antibodies and antibody fragments from serumbased culture. Features and benefits Use for oriented capture of antibody fragments Ready-to-use, eliminating the need to develop immobilization and regeneration conditions, saving time and effort Captures a wide range of antibody fragments, such as Fabs; single-chain variable fragments (scFv); kappa light chain subtypes 1, 3, and 4; and domain antibodies (dAbs Product Quantity Product code Series S sensor chip protein L Pack of 1 29205138 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 82 Features and benefits Versatile: immobilizes biotinylated peptides, proteins, nucleic acids, or carbohydrates Ready-to-use: streptavidin is pre-immobilized, ready for fast, high-affinity ligand capture Robust: streptavidin-biotin is one of the strongest noncovalent interactions in nature Noncovalent: a convenient alternative for ligands refractory to covalent immobilization Standardized ligand orientation: controlled biotinylation enables oriented capture Product Quantity Product code Series S sensor chip SA Pack of 3 BR100531 Series S sensor chip SA Pack of 1 29104992 Series S sensor chip SA Pack of 10 29699621 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Series S sensor chip SA This high-affinity streptavidin (SA) sensor chip immobilizes biotinylated molecules for surface plasmon resonance (SPR) interaction analysis on Biacore systems. Sensor Chip SA provides a surface of streptavidin covalently attached to a carboxymethyl dextran matrix. Dissociation of biotinylated ligands from the surface of the chip is generally negligible during a Biacore analysis because the affinity of streptavidin for biotin is extremely high, with an equilibrium dissociation constant of about 10 to 15 M. High binding capacity, reproducibility, and chemical resistance give excellent performance over a broad range of applications. The surface of sensor chip SA is resistant to 1-minute pulses of many agents commonly used in SPR sensor chip washing and regeneration protocols. Biotin has become a popular molecular tag for capture and interaction studies because it is stable and rarely interferes with target activity or structure due to its small size. Biotin can be conveniently conjugated to a wide range of target molecules by chemical or enzymatic means. Applications for sensor chip SA are therefore broad. Examples include: characterizing PROTAC ternary complex formation screening of endocrine-disrupting chemicals (3) elucidating DNA- and RNA- mediated mechanisms of catechins in cancer prevention (4) 83 Series S sensor chip NTA Biacore sensor chip NTA captures histidine-tagged molecules by metal chelation for subsequent surface plasmon resonance (SPR) interaction analysis. Histidine is the most widely used molecular tag today. Sensor Chip NTA can be used to capture many different types of polyhistidinetagged proteins for subsequent SPR assays. Interaction of immobilized proteins with a wide range of analyte molecules can be studied, from low-molecular-weight compounds to large proteins. For experiments where low-molecular-weight analytes are studied, sensor chip NTA is the first choice. Sensor chip NTA consists of carboxymethylated dextran with covalently immobilized nitrilotriacetic acid (NTA). The NTA molecule chelates metal ions such as nickel (Ni2+), creating coordination sites that bind to polyhistidine tags on recombinant proteins and other biomolecules. Noncovalent capture of ligands, for example by metal ion chelation, has a number of advantages over covalent immobilization. Tagged proteins can be captured directly from crude cell extracts or culture medium, requiring little or no sample preparation. Because fresh ligand is captured for each cycle, the same chip surface can be used for analysis interactions involving different ligands. Regeneration conditions are generic and therefore the need for assay development is minimized. In addition, capture generates a directed structural orientation of the protein on the surface, potentially offering optimal site exposure. Features and benefits Versatile: captures many types of histidine-tagged molecules Ready-to-use: NTA is pre-immobilized, ready for nickel loading and ligand capture Convenient: compatible NTA reagent kit provides nickel ion and regeneration solutions Easily regenerated: EDTA injection efficiently removes metal ions to regenerate the sensor Chelation capture: simpler assay development, potential to standardize target orientation Product Quantity Product code Series S sensor chip NTA, pack of 3 Pack of 3 BR100532 Series S sensor chip NTA, pack of 1 Pack of 1 28994951 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 84 His capture kit type 2 His capture kit, type 2 is designed for capture of histidine-tagged proteins to an anti-histidine antibody and provides an alternative to capture on nickel-chelated nitrilotriacetic (NTA) groups. The anti-histidine antibody is a monoclonal antibody directed against polyhistidine tags in the C- or N-terminus of a protein. Depending on the nature of the interacting molecules and the micro-environment around the histidine tag, the affinity of the capture antibodies will vary. Features and benefits Enables capture of histidine-tagged ligands; physiological conditions can be used during the coupling Optimized and verified assay minimizing assay development Protocol and required solutions provided Product Quantity Product code His capture kit, type 2 1 29234602 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 85 Product Quantity Product code Biotin CAPture kit, series S 1 28920234 Biotin CAPture kit series S Kit containing reagents and sensor chip for reversible capture of biotinylated molecules. Series S sensor chip CAP Biotin CAPture reagent Regeneration stock 1 Regeneration stock 2 The ligand is captured on to series S sensor chip CAP via Biotin CAPture reagent, which is a modified streptavidin. Series S sensor chip CAP should only be used together with biotin CAPture reagent. Regeneration of the surface after each analysis cycle removes biotin CAPture reagent as well as the ligand and any bound analyte. Fresh biotin CAPture reagent is attached to the surface for each cycle. Sensor ship CAP and biotin CAPture reagent contain deoxyribo-oligonucleotides. Therefore, the kit is not suitable for work with DNA-binding proteins or enzymes that degrade DNA. The ligand capture capacity of sensor chip CAP and biotin CAPture reagent is typically about 1500 to 3000 RU for a ligand with Mr 150 000. Biotin CAPture kit has been optimized for an assay temperature of 25°C. Features and benefits Study interactions without the need to know or establish immobilization methods or regeneration protocols; only biotinylation of the ligand is required Study unstable ligands or ligands for which no regeneration can be established Maximize economy and efficiency when working with multiple ligands For most types of interaction analyses in Biacore systems Sufficient for 100 regenerations Download Download our poster on reversible capture of biotinylated molecules for Biacore analysis using Biotin CAPture kit. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 86 Gluthatione-S transferase (GST) capture kit Reagents for site-directed affinity capture of GST fusion proteins in biomolecular interaction analyses using Biacore systems. Goat anti-GST antibody, 0.8 mg/mL in 75 µL coupling solution, 5 mL positive control: recombinant GST (Schistosoma japonicum), 0.2 mg/mL in 100 µL HBS-EP regeneration solution, 70 mL. The use of these products in Biacore systems requires an amine coupling kit with sensor chip CM5, CM4, CM3, or C. Not for use with Biacore Flexchip Anti-GST antibody is suitable for immobilization on carboxylderivatized sensor chips using amine coupling kit and the included immobilization buffer. Regeneration solution is used for regeneration of the surface by removal of the captured fusion protein. Recombinant GST is used in preparation of the surface and may also be used in assay development work to check that the sample does not bind to GST. Features and benefits For most types of interaction analyses in Biacore systems Sufficient for 20 immobilizations and up to 600 regenerations Product Quantity Product code GST capture kit 1 BR100223 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 87 Human antibody capture kit type 2 Human antibody capture kit, type 2 is intended for capture of human or humanized IgG antibodies as ligands in biomolecular interaction analyses. Anti-human IgG (Fc) antibody is a monoclonal mouse antihuman IgG (Fc) antibody of IgG1 subclass, prepared by affinity chromatography on protein A. The antibody recognizes an epitope within the CH₂ domain in human IgG Fc of all subclasses. It is also known to bind to monkey (cynomolgus) and rabbit IgG. Features and benefits Binds an epitope in CH₂ domain of human IgG (all subclasses recognized) Little or no cross-reactivity towards other human Ig isotypes No, or very low, cross-reactivity with IgG from other species, except other primates and rabbit Very stable binding — little or no dissociation of captured antibodies Product Quantity Product code Human antibody capture kit, type 2 1 29234600 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 88 Product Quantity Product code Human Fab capture kit, type 2 1 29234601 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Human Fab capture kit type 2 Human Fab binder is a mixture of two monoclonal antibodies recognizing, respectively, kappa and lambda subtypes of human Fab fragment light chains. It binds specifically to the Fab region of human antibodies and no binding to the Fc region has been observed. It recognizes most subtypes of both kappa and lambda light chains and does not cross-react with Fab from other species. In addition, no nonspecific binding from cell lysate or cell culture medium has been observed. The main applications of human Fab capture kit, type 2 are screening and characterization of human Fab fragments generated from phage display (purified or directly from crude cell lysates or periplasmic extract). Features and benefits Early selection of binders enabled through kinetics screening by high resolution off-rate ranking Broad framework specificity; kappa and lambda types of human Fab fragments captured Excellent assay performance; fab binder with high capture efficiency and stability 89 Mouse antibody capture kit type 2 Mouse antibody capture kit, type 2 is intended for capture of mouse antibodies as ligands in various biomolecular interaction analyses. Anti-mouse antibodies are suitable for immobilization on carboxyl derivatized surfaces using amine coupling kit and the included immobilization buffer. The regeneration solution is used for regeneration of the surface. Anti-mouse antibodies consist of polyclonal rabbit anti-mouse immunoglobulin antibodies reacting with all IgG subclasses (IgG1, IgG2a, IgG2b, and IgG3), IgA and IgM. Reaction with other mouse immunoglobulin classes has not been investigated, but the antibodies are likely to react with these via their light chains. Features and benefits Binds all IgG subclasses (IgG1, IgG2a, IgG2b, and IgG3), IgA, and IgM. Very stable binding with no or very low dissociation of captured antibodies. Product Quantity Product code Mouse antibody capture kit, type 2 1 29215281 Read more Read more about protein purification methods: ‘Your guide to protein purification basics, tips, and tricks, and more’. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 90 Reagent purification Immunoassay reagent characterization High-quality antibodies or antibody fragments are key components in immunoassays. For a successful diagnostic assay, a detailed analysis of your reagents’ binding properties and of the behavior of the detector molecules will guide your selection and support your QC over the complete life span. Biacore surface plasmon resonance (SPR) systems aid you and provide answers to reagent specificity, active analyte concentration, and reagent affinity and kinetics (i.e., how strong and how fast or slow a binding complex forms and decays). Learn more Learn more about Biacore SPR — surface plasmon resonance interaction analysis. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 91 How Biacore surface plasmon resonance is shaping today’s assay development For over 40 years, immunoassays have been used by hospitals, laboratories, and researchers to rapidly diagnose a wide variety of health, disease, and drug treatment conditions. Information gained by these tests has helped to identify presymptomatic disease and improve therapeutic options for patients. In life science research, immunoassays have been used to pinpoint hits, quantitate protein expression levels and sample concentration, and unravel complex molecular interactions to advance biological study. In industry, immunoassays have been used to detect contaminants in our food and water. These quick and accurate tests fall within the broader category of methods frequently described as binding assays (Fig 1). Immunoassay developers are constantly striving to create tests that address unmet diagnostic needs. Doing so would enable them to gain a first-to-market advantage over competitors. But no matter how innovative an assay is, if it does not consistently perform as promised, it is unlikely to generate repeat purchase and will ultimately fail. A critical prerequisite for good assay performance on any technology platform is the design, production, and characterization of high-quality reagents that are highly selective and sensitive. There are challenges in sourcing reagents that must be addressed in order to develop novel, precise, and fit-for-purpose immunoassays. The following guidelines highlight some critical considerations in developing immunoassay reagents. Steps for avoiding common pitfalls are outlined for reagent developers and for those who develop the assays that depend upon qualified reagents. Fig 1. Conventional sandwich immunoassay. Sandwich Read more Read more about immunoassay reagent development using Biacore SPR. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 92 Product Quantity Product code Protein A Mag Sepharose 500 µl 28944006 Protein A Mag Sepharose 4 × 500 μl 28951378 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Protein A Mag Sepharose™ beads Protein A Mag Sepharose magnetic beads are designed to simplifying enrichment of target proteins via immunoprecipitation or pull-down assays used to detect physical interactions between two or more proteins. Protein A Mag Sepharose beads combine well-proven enrichment methods with the advantages of using magnetic beads, making protein A Mag Sepharose beads highly suited for small-scale sample preparation. Two protocols are available for protein enrichment: the cross-link protocol (linking proteins together via chemical bonds between amino acids) and the classic protocol for protein interaction. Both use simple elution conditions optimized for electrophoresis and mass spectrometry (MS) analysis. Features and benefits Visible and dense Sepharose-resin-based magnetic beads with protein A as the ligand make it easy to spot and collect bound target protein Nonadherent beads eliminate smearing effects and aggregate formation Can be used without detergents Simple capture of target protein in small or large sample volumes (low microliter to high milliliter scale) Optimized capacity for enrichment or immunoprecipitation of target proteins requiring only low amounts of antibody needed Flexible protocols with elution conditions optimized for both electrophoresis and MS analysis 93 Product Quantity Product code Protein G Mag Sepharose 500 µl 28944008 Protein G Mag Sepharose 4 × 500 μl 28951379 Protein G Mag Sepharose beads Protein G Mag Sepharose beads simplify enrichment of target proteins by immunoprecipitation techniques, protein antigen precipitation from solutions using antibodies, or pull-down applications as an in vitro technique to detect physical interactions. Protein G Mag Sepharose beads combine enrichment methods with magnetic beads for small-scale sample preparation. These magnetic beads are coated with Sepharose, a beaded-form of agarose, with protein G as the ligand due to its high affinity for the Fc region of IgG antibodies. Two protocols are available for protein enrichment: the cross-link protocol (linking proteins together via chemical bonds between amino acids) and the classic protocol for protein interaction. Both are simple elution conditions optimized for electrophoresis and MS. Features and benefits Visible and dense Sepharose-resin-based magnetic beads with protein G ligand making it easy to spot and collect bound target protein Nonadherent beads eliminate smearing effects and aggregate formation Can be used without detergents Simple capture of target protein in small or large sample volumes (low microliter to high milliliter scale) Optimized capacity for enrichment/immunoprecipitation of target proteins — only low amounts of antibody needed Flexible protocols with elution conditions optimized for both electrophoresis and MS analysis Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 94 Product Quantity Product code Streptavidin Mag Sepharose 5 × 1 mL 10% slurry 28985799 Streptavidin Mag Sepharose 2 × 1 ml 10% slurry 28985738 Streptavidin Mag Sepharose beads Streptavidin Mag Sepharose magnetic beads are designed for simple and efficient enrichment of target proteins through immunoprecipitation and purification of biotinylated biomolecules. Features and benefits High-capacity, small-scale purification of proteins from cell lysates and biological fluids High purity and yield Easy parallel screening of proteins with high repeatability Simple enrichment of proteins from small or large sample volumes (low microliter to high milliliter scale) Utilizes strong interaction between biotin and streptavidin ligand immobilized on magnetic beads. Available in two pack sizes: 2 × 1 mL 10% medium slurry and 5 × 1 mL 10% medium slurry. 1 mL of 10% medium slurry is equal to 100 μL sedimented medium, sufficient for 20 purification runs. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 95 Product Quantity Product code His Mag Sepharose Ni 5 × 1 mL 28967390 His Mag Sepharose Ni 100 mL 29104065 His Mag Sepharose Ni 2 × 1 mL 28967388 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection His Mag Sepharose Ni beads His Mag Sepharose Ni beads are magnetic beads designed for rapid, small-scale purification and screening of histidine-tagged proteins from different sources. Lower concentrations of histidine-tagged proteins are commonly used to wash resins prior to elution. Mag Sepharose bead format has excellent properties for small-scale experiments. This product is based on immobilized metal ion affinity chromatography (IMAC) Sepharose, incorporating magnetite and immobilized nickel. Features and benefits Easy-to-use, visible magnetic beads ensures reliable collection of bound histidine-tagged proteins in purification procedures High purity and yield Easy parallel screening with high repeatability High density of beads ensures rapid capture from magnetic devices Scalable-simple capture of target proteins from small µL to high mL sample volumes 96 Product Quantity Product code His Mag Sepharose excel 10 × 1 mL 17371222 His Mag Sepharose excel 5 × 1 mL 17371221 His Mag Sepharose excel 2 × 1 mL 17371220 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection His Mag Sepharose Excel beads His Mag Sepharose Excel beads are magnetic beads designed for rapid, small-scale purification and screening of his-tagged proteins consisting of an immobilized metal ion affinity chromatography (IMAC) resin precharged with nickel ions that are strongly bound to a chelating ligand. Small-scale capture, purification, and screening of histidine-tagged proteins that are expressed and secreted into supernatants from eukaryotic cells such as insect cells or Chinese Hamster Ovary (CHO) cells in HyClone™ HyCell™ CHO cell culture media. Features and benefits Simplifies small-scale purification and expression screening of histidine-tagged proteins secreted in eukaryotic cell culture Robust parallel screening with high reproducibility Increase target protein yield and decrease degradation through reduced and simplified sample handling Capture histidine-tagged protein from different sample volumes (low microliter to high milliliter scale) Visible and dense Sepharose-resin-based magnetic beads give increased simplicity for handling High density of beads ensures rapid capture by magnetic devices 97 Product Quantity Product code NHS Mag Sepharose 4 × 500 μL 28951380 NHS Mag Sepharose 500 μL 28944009 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection NHS Mag Sepharose beads NHS Mag Sepharose magnetic beads are designed for pull-down techniques involving isolation of a protein through adsorption of complex samples onto beads, enabling rapid capture and enrichment of selected proteins based on affinity. Flexible protocols with elution conditions optimized for both electrophoresis and MS analysis of antibodies or other proteins with the desired affinity are covalently coupled to the medium via primary amines for purpose of target proteins of interest. Features and benefits Visible and dense Sepharose-resin-based magnetic beads with N-hydroxysuccinimide ligand for easy coupling of biomolecules containing a free amine group Nonadherent beads eliminate smearing effects and aggregate formation Can be used without detergents Simple capture of target protein in small or large sample volumes (low microliter to high milliliter scale) Optimized capacity for enrichment or immunoprecipitation of target proteins, only low amounts of antibody needed 98 Product Quantity Product code HisTrap HP 5 × 5 mL 17524802 HisTrap HP 5 × 1 mL 17524701 HisTrap HP 1 × 5 mL 17524801 HisTrap HP 1 × 1 mL 29051021 Affinity chromatography HisTrap™ HP His tag protein purification columns HisTrap™ HP prepacked columns provide a fast, convenient, and reproducible format for preparative purification of histidine-tagged (His-tag) recombinant proteins. Features and benefits High resolution: due to small bead size (average bead size is 34 µm) High binding capacity: at least 40 mg His tag protein per mL resin, for high yields Negligible nickel leakage: helps retain activity and minimize protein precipitation Broad reagent compatibility: suitable for use with a wide range of reducing agents, detergents, denaturants, and other additives Convenient HiTrap™ format: 1 mL and 5 mL columns compatible with syringes, pumps, or chromatography systems such as ÄKTA™ systems or other FPLC systems His tag protein purification by IMAC is both popular and highly efficient. HisTrap HP columns are packed with Ni Sepharose high performance (HP) affinity resin. This resin consists of highly cross-linked agarose beads to which a chelating group has been coupled. The chelating group is precharged with nickel, which selectively retains proteins with exposed histidine groups. HisTrap HP IMAC resin has a binding capacity that exceeds 40 mg/mL, resulting in high protein recovery. The small bead size (34 µm) contributes to a purer purification compared to nickel resins of larger bead size. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 99 Product Quantity Product code HisTrap FF 5 × 5 mL 17525501 HisTrap FF 5 × 1 mL 17531901 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection HisTrap FF columns HisTrap FF column is a ready-to-use column, prepacked with precharged Ni Sepharose 6 fast flow resin for preparative purification of histidine-tagged recombinant proteins by immobilized metal ion affinity chromatography (IMAC). Features and benefits High binding capacity, approx. 40 mg/mL resin Negligible leakage of Ni2+ Prepacked columns offer reliable and convenient time-saving purification of histidine-tagged recombinant proteins Compatible with a wide range of reducing agents, detergents, denaturants, and other additives Ni Sepharose 6 fast flow resin consists of 90 µm beads of highly cross-linked agarose to which a chelating ligand has been immobilized. This chelating ligand is charged with Ni2+ ions, the first-choice metal ion for purifying most histidine-tagged proteins. The negligible leakage of Ni2+ ions from the matrix ensures reliable capture of histidine-tagged proteins in repeated IMAC purifications. The high flow rates made possible by the Ni Sepharose 6 fast flow resin matrix make HisTrap FF columns well suited for small scale-up. Capacity can be increased by connecting columns in series. HiTrap IMAC FF columns are the product of choice when charging the resin with different metal ions for optimization of purification protocols. 100 Product Quantity Product code HisTrap excel 5 × 5 mL 17371206 HisTrap excel 5 × 1 mL 17371205 HisTrap excel 1 × 1 mL 29048586 HisTrap excel columns are made of biocompatible polypropylene. The columns are delivered with a stopper on the inlet and a snapoff end on the outlet. HisTrap excel columns can be operated either with a peristaltic pump or with chromatography systems such as ÄKTA systems. The columns cannot be opened or refilled. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection HisTrap excel columns HisTrap excel columns are prepacked with Ni Sepharose excel affinity resins for capture and purification of histidine-tagged proteins secreted into eukaryotic cell culture supernatants by immobilized metal ion affinity chromatography (IMAC). Features and benefits Load eukaryotic cell culture samples containing secreted histidine-tagged proteins directly with retained binding capacity Increase target protein yield and decrease degradation through reduced and simplified sample handling HisTrap excel columns allow direct purification of cell-free, unclarified samples HisTrap excel columns are ready-to-use IMAC columns prepacked with Ni Sepharose excel resin. The design of the columns in combination with the specific properties of the resin enables fast and convenient purifications. The special type of filter in the top and bottom of the columns makes it possible to load large volumes of cell-free, unclarified samples directly on the columns without causing back pressure problems. This time-saving property helps prevent degradation and loss of sensitive target proteins. 101 Product Quantity Product code HisTrap FF crude columns 5 × 5 mL 17528601 HisTrap FF crude columns 5 × 1 mL 11000458 HisTrap FF crude columns 1 × 1 mL 29048631 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection HisTrap FF crude columns HisTrap FF crude columns are high binding capacity IMAC columns for His-tagged protein purification when scalability is needed. High binding capacity: binds up to 40 mg His-tag protein/mL resin Excellent flow properties: great performance even at high flow rates Optimized for unclarified samples: short purification time can minimize potential negative effects such as degradation and oxidation of proteins HiTrap column format: compatible with syringes, pumps, or chromatography systems such as ÄKTA systems or other FPLC systems HisTrap FF crude is a ready-to-use column, prepacked with precharged Ni Sepharose 6 fast flow resin. After thorough cell disruption, it is possible to load the unclarified lysate on the column, without precentrifugation and filtration. If you plan to load unclarified samples, it is important to ensure that lysis is as complete as possible. We recommend adding lysozyme and DNase I, followed by mechanical lysis (e.g., by sonication) for a longer duration than typically performed. 102 Product Quantity Product code GSTrap FF 5 × 5 mL 17513102 GSTrap FF 5 × 1 mL 17513001 GSTrap FF 2 × 1 mL 17513002 GSTrap FF 1 × 5 mL 17513101 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection GSTrap™ FF columns GSTrap™ FF columns are prepacked glutathione Sepharose fast flow columns for fast, convenient, one-step purification of glutathione S-transferase (GST) tagged proteins. Features and benefits Fast and simple one-step purification of GST-tagged proteins Mild elution conditions preserving protein antigenicity and function Easy scale-up by connecting the columns in series Sepharose 4 fast flow resin provides good flow properties GST-tagged proteins can be purified directly from pretreated bacterial lysates using GSTrap FF columns. GST-tagged proteins are eluted under mild, nondenaturing conditions using reduced glutathione. The purification process preserves protein antigenicity and function. If desired, cleavage of the protein from GST can be achieved using a site-specific protease whose recognition sequence is located immediately upstream from the multiple cloning site on the pGEX plasmids. GST-tagged proteins can be detected using colorimetric or immunological methods. The resin, glutathione Sepharose 4 fast flow, is also available as lab packages and is an excellent choice for scale-up. The columns can be operated with a syringe, a peristaltic pump, or liquid chromatography systems such as ÄKTA systems or FPLC systems. 103 Product Quantity Product code GSTrap HP 5 × 5 mL 17528202 GSTrap HP 5 × 1 mL 17528101 GSTrap HP 1 × 5 mL 17528201 GSTrap HP columns GSTrap HP columns are prepacked with glutathione Sepharose high performance resin and provide high-resolution, one-step purifications of glutathione S-transferase (GST) tagged proteins. Features and benefits High resolution purification of GST-tagged proteins Mild elution conditions preserving protein antigenicity and function Fast, one-step purification Easy to use with syringes, pumps, or chromatography systems GSTrap HP columns are prepacked with glutathione Sepharose high performance resin, which provides high-resolution purifications and thus a more concentrated sample. GST-tagged proteins are purified from pretreated bacterial lysates by affinity chromatography using immobilized glutathione, such as glutathione Sepharose high performance resin. GST-tagged proteins are bound to the affinity resin, and impurities are removed by washing with binding buffer. GST-tagged proteins are eluted under mild, nondenaturing conditions using reduced glutathione. The purification process preserves protein antigenicity and function. If desired, cleavage of the protein from GST can be achieved using a site-specific protease whose recognition sequence is located immediately upstream from the multiple cloning site on the pGEX plasmids. GST-tagged proteins can be detected using colorimetric or immunological methods. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 104 Product Quantity Product code GSTrap 4B 5 × 5 mL 28401748 GSTrap 4B 1 × 5 mL 28401747 GSTrap 4B 1 × 1 mL 29048609 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection GSTrap 4B columns GSTrap 4B columns are prepacked glutathione Sepharose 4B columns for convenient, high capacity one-step purification of glutathione S-transferase (GST) tagged proteins. Features and benefits High binding capacity Mild elution conditions preserving protein antigenicity and function Easy one-step purifications of glutathione S-transferase (GST) tagged proteins resulting in high purity GSTrap 4B columns are designed to be used with syringes, pumps, or chromatography systems such as ÄKTA systems GST-tagged proteins can be purified directly from pretreated bacterial lysates using GSTrap 4B columns. GST-tagged proteins are eluted under mild, nondenaturing conditions using reduced glutathione. The purification process preserves protein antigenicity and function. If desired, cleavage of the protein from GST can be achieved using a site-specific protease whose recognition sequence is located immediately upstream from the multiple cloning site on the pGEX plasmids. GST-tagged proteins can be detected using colorimetric or immunological methods. The resin, glutathione Sepharose 4B, is also available as lab packages and is a good choice for scale-up. The columns can be operated with a syringe, a peristaltic pump, or a liquid chromatography system such as an ÄKTA system or a FPLC system. 105 Product Quantity Product code MabSelect resin 25 mL 17519901 MabSelect resin 200 mL 17519902 MabSelect resin 1 L 17519903 MabSelect resin 5 L 17519904 MabSelect resin 10 L 17519906 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection MabSelect™ antibody purification chromatography resin MabSelect™ resin is a BioProcess™ protein A resin for capturing monoclonal antibodies (mAbs) from large sample volumes. Features and benefits High-flow agarose matrix allows for high flow velocities at production scale enabling the processing of more than 10 000 L of feed in one working day. Oriented coupling of the ligand and optimized matrix gives high dynamic binding capacity, which reduces resin volume requirements. Low nonspecific binding resulting in low levels of impurities in the product eluate pool. BioProcess resin supported for industrial applications and wellestablished in approved processes. Resin fulfills industrial demands for security of supply, robust performance, and regulatory support. The resin is based on a highly cross-linked agarose matrix with a recombinant protein A ligand. This ligand is engineered to favor an oriented coupling resulting in an affinity resin with enhanced binding capacity for IgG. The hydrophilic nature of the base matrix ensures low levels of nonspecific binding leading to low levels of host-cell-derived impurities in the elution pool. MabSelect resin is optimized for high capacity, throughput, and to deliver a product pool that is high in purity and yield. 106 Product Quantity Product code HiTrap MabSelect PrismA 5 × 5 mL 17549854 HiTrap MabSelect PrismA 5 × 1 mL 17549852 HiTrap MabSelect PrismA 1 × 5 mL 17549853 HiTrap MabSelect PrismA 1 × 1 mL 17549851 HiTrap MabSelect PrismA™ protein A columns These HiTrap columns are prepacked with MabSelect PrismA™ protein A chromatography resin. This affinity resin in the packed column has been improved with an optimized high-flow agarose base matrix and a genetically engineered protein ligand, allowing efficient cleaning between monoclonal antibody purification runs. This efficient cleaning allows future demands in monoclonal antibody purification to be met including processing of many bispecific antibodies. Features and benefits Enhanced dynamic binding capacity compared with other protein A resins Excellent alkaline stability enables efficient cleaning and sanitization using 0.5 to 1.0 M NaOH Convenient HiTrap format for easy connection to a syringe, a peristaltic pump, or chromatography systems such as an ÄKTA system for convenient process optimization Easily measure protein A leakage using the PrismA ELISA™ kit specifically designed for use with MabSelect PrismA resins The enhanced alkaline stability of MabSelect PrismA resin also provides opportunities for more robust mAb purification using periodic counter-current chromatography. The possibility for cleaning using 1 M NaOH increases bioburden control for continuous processes, which can be exposed to days or weeks of feed harvest. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 107 Product Quantity Product code MabSelect SuRe 25 mL 17543801 MabSelect SuRe 200 mL 17543802 MabSelect SuRe 1 L 17543803 MabSelect SuRe 5 L 17543804 MabSelect SuRe 10 L 17543805 MabSelect SuRe™ antibody purification resin The recombinant protein A ligand of MabSelect SuRe resin has been engineered to withstand 0.1 to 0.5 M NaOH, enabling efficient cleaning of the chromatography resin. Features and benefits In process-scale applications, this NaOH tolerance extends the lifetime of the resin and allows for the use of rigorous and cost-effective CIP and sanitization protocols. In research applications, this NaOH tolerance enables the purification of multiple antibody types with minimized cross-contamination risk. Protein ligand designed for enhanced protease resistance compared with unmodified protein A leads to lower ligand leakage. Meets industrial demands in terms of comprehensive technical and regulatory support and security of supply Animal-component-free raw materials and manufacturing process This resin for monoclonal antibody (mAb) purification contains a rigid, high-flow agarose matrix that delivers excellent pressure/flow properties. Like many protein A chromatography resins from Cytiva, MabSelect SuRe resin uses oriented ligand coupling for excellent binding capacity. It exhibits low nonspecific protein binding, thereby decreasing impurity levels in the eluate pool. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 108 Product Quantity Product code MabSelect PrismA Protein A resin 25 mL 17549801 MabSelect PrismA Protein A resin 200 mL 17549802 MabSelect PrismA Protein A resin 1 L 17549803 MabSelect PrismA Protein A resin 5 L 17549804 MabSelect PrismA Protein A resin 10 L 17549805 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection MabSelect PrismA Protein A chromatography resin MabSelect PrismA affinity chromatography resin has been improved with an optimized high-flow agarose base matrix and a genetically engineered protein A-derived ligand. This resin allows future demands in mAb purification to be met, including processing of many bispecific antibodies. Features and benefits Enhanced dynamic binding capacity allows high mass throughput of processed mAb per resin volume unit. Excellent alkaline stability enables efficient cleaning and sanitization using 0.5 to 1.0 M NaOH for improved process economy, bioburden control, and robustness. Covered by a comprehensive security of supply program, including dual sources of the agarose base matrix and protein A ligand. The PrismA protein A ligand is also available in HiTrap Fibro™ and HiScreen Fibro™ PrismA chromatography units for ultrafast purification of mAbs. Easily measure protein A leakage using the PrismA ELISA kit specifically designed for use with MabSelect PrismA resins. The enhanced alkaline stability of MabSelect PrismA resin also provides opportunities for more robust mAb purification using periodic counter-current chromatography. The possibility for cleaning using 1 M NaOH increases bioburden control for continuous processes, which can be exposed to days or weeks of feed harvest. 109 Product Quantity Product code HiScreen MabSelect SuRe 1 × 4.7mL 28926977 HiScreen MabSelect SuRe protein A columns The HiScreen MabSelect SuRe column uses protein A chromatography for antibody purification. This protein A column is prepacked with MabSelect SuRe affinity resin and is part of our process development platform. These packed columns are used for method optimization and parameter screening for capture of monoclonal antibodies. Features and benefits The novel, alkali-tolerant recombinant protein A ligand of MabSelect SuRe resin allows cleaning-in-place (CIP). Prepacked, ready-to-use columns for convenient chromatography process development. 10 cm bed height of HiScreen columns is designed to allow method optimization and parameter screening. Easily connected in series to achieve 20 cm bed height. Small bed volume gives fast results and minimal sample/buffer consumption. Reproducible results, scalable to BioProcess columns packed with the same chromatography resin using the same linear fluid velocity. The small column volume of 4.7 mL and bed height of 10 cm make HiScreen columns excellent tools for method optimization, parameter screening, robustness testing, and convenient scale-up of the protein purification process. Process fluid velocities can be applied because the 10 cm bed height gives enough residence time, and the results can then serve as basis for linear process scale-up. Protein A resins prepacked in HiScreen MabSelect SuRe chromatography columns are also available in filter plates and as bulk packs, which makes it possible to use the same resins for development work, pilot studies, and clinical production. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 110 Product Quantity Product code MabSelect SuRe pcc 25 mL 17549101 MabSelect SuRe pcc 200 mL 17549102 MabSelect SuRe pcc 1 L 17549103 MabSelect SuRe pcc 5 L 17549104 MabSelect SuRe pcc 10 L 17549105 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection MabSelect SuRe pcc protein A resin MabSelect SuRe pcc protein A resin is an affinity chromatography resin for purification of monoclonal antibody and Fc-fusion proteins. The high resin capacity at short residence times makes it an excellent choice for periodic counter-current (pcc) chromatography in monoclonal antibody (mAb) purification. The alkaline-stable, recombinant protein ligand allows for efficient resin cleaning. Features and benefits High dynamic binding capacity (e.g., ~ 60 g IgG/L resin at 2.4 min residence time) High productivity through capture of large mass of monoclonal antibody in a short period of time Covered by a comprehensive security of supply program, including dual sources of the agarose base matrix and protein A ligand The alkali-stabilized, protein A-derived ligand gives the resin greater stability than conventional protein A-based resins under the alkaline conditions used in CIP protocols. Consequently, the possibility of cleaning with cost-effective reagents such as NaOH will improve process economy. In addition, the use of NaOH for cleaning efficiently inactivates bacteria, mold, and yeast, with improved product quality as a result. 111 Product Quantity Product code HiScreen MabSelect SuRe LX 1 × 4.7mL 17547415 HiScreen MabSelect SuRe LX protein A columns The HiScreen MabSelect SuRe LX column uses protein A chromatography for antibody purification. This protein A column is prepacked with MabSelect SuRe LX affinity resin and is part of our process development platform. These packed columns are an excellent choice for method optimization and parameter screening for capture of monoclonal antibodies when a high dynamic binding capacity is needed. Features and benefits The alkali-tolerant recombinant protein A ligand of MabSelect Sure LX resin allows CIP, and the resin gives excellent dynamic binding capacity. Prepacked, ready-to-use columns for convenient chromatography process development. 10 cm bed height of HiScreen columns is designed to allow method optimization and parameter screening. Easily connected in series to achieve 20 cm bed height. Small bed volume gives fast results and minimal sample/buffer consumption. Reproducible results, scalable to BioProcess columns packed with the same chromatography resin using the same linear fluid velocity. The small column volume of 4.7 mL and bed height of 10 cm make HiScreen columns excellent tools for method optimization, parameter screening, robustness testing, and convenient scaleup of the protein purification process. Process fluid velocities can be applied because the 10 cm bed height gives enough residence time, and the results can then serve as basis for linear process scale-up. Protein A resins prepacked in HiScreen MabSelect SuRe LX chromatography columns are also available in filter plates and as bulk packs, which makes it possible to use the same resins for development work, pilot studies, and clinical production. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 112 Product Quantity Product code HiScreen MabSelect PrismA 1 × 4.7mL 17549815 HiScreen MabSelect PrismA protein A columns This protein A column is prepacked with MabSelect PrismA chromatography resin. This affinity resin has been improved with an optimized high-flow agarose base matrix and a genetically engineered protein ligand, allowing efficient cleaning between runs. This efficient cleaning allows future demands in monoclonal antibody purification to be met, including processing of many bispecific antibodies. Features and benefits Enhanced dynamic binding capacity compared with other protein A chromatography resins Excellent alkaline stability enables efficient cleaning and sanitization using 0.5 to 1.0 M NaOH Convenient method optimization and parameter screening enabled by the 10 cm bed height for efficient chromatography process development Easily measure protein A leakage using the PrismA ELISA kit specifically designed for use with MabSelect PrismA resins The enhanced alkaline stability of MabSelect PrismA resin also provides opportunities for more robust mAb purification using periodic counter-current chromatography. The possibility for cleaning using 1 M NaOH increases bioburden control for continuous processes, which can be exposed to days or weeks of feed harvest. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 113 Product Quantity Product code Capto DEAE 25 mL 17544310 Capto DEAE 100 mL 17544301 Capto DEAE 1 L 17544303 Capto DEAE 5 L 17544304 Capto DEAE 10 L 17544305 Capto DEAE 60 L 17544360 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Ion exchange chromatography Capto™ DEAE ion exchange chromatography resin Capto™ DEAE is a weak anion exchanger resin for large-scale manufacturers. Meets demands for fast, cost-effective capture and intermediate protein purification. Features and benefits Weak anion exchanger for high-productivity capture and intermediate purification when high volume throughput is essential High dynamic binding capacity at high flow raises productivity High-volume throughput cuts process times Cost-effective processing with smaller unit operations Resin fulfills industrial demands for security of supply, robust performance, and regulatory support Suited to high volume applications Capto DEAE resin is designed for capture and intermediate purification of proteins from large feed volumes. It is composed of a rigid, high-flow agarose matrix modified with dextran surface extenders and a weak diethylaminoethyl (DEAE) anion exchanger. 114 Product Quantity Product code Capto Q ImpRes 25 mL 17547010 Capto Q ImpRes 100 mL 17547002 Capto Q ImpRes 1 L 17547003 Capto Q ImpRes 5 L 17547004 Capto Q ImpRes 10 L 17547004 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Capto Q ImpRes ion exchange chromatography resin By combining high-flow characteristics of Capto resins with a small particle size, Capto Q ImpRes resin delivers high resolution and excellent pressure-flow properties. Features and benefits High-resolution, intermediate purification and polishing based on the well-established Capto resin platform with traditional ligands Flexible process design due to large operational window of flow rates and bed heights High-throughput purifications easy to optimize and scale-up Higher manufacturing productivity enables improved process economy Resin fulfills industrial demands for security of supply, robust performance, and regulatory support Capto Q ImpRes resin is a strong anion exchanger for the highthroughput, intermediate purification and polishing steps of a wide range of biomolecules. This chromatography resin is part of an expanded high-resolution platform based on the high-flow agarose Capto resom product line. The ability to run at higher flow rates and higher bed heights also increases flexibility in process design. 115 Product Quantity Product code Capto Q XP 25 mL 17547301 Capto Q XP 100 mL 17547302 Capto Q XP 1 L 17547303 Capto Q XP 5 L 17547304 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Capto Q XP ion exchange chromatography resin Capto Q XP resin is designed for capture and intermediate purification of large biomolecules, such as immunoglobulins IgG, IgM, IgA, in downstream processes. Features and benefits Efficient industrial-scale purification based on the wellestablished Capto resin platform with traditional ligands Flexible process design due to large operational window of flow rates and bed heights High throughput to improve manufacturing productivity and process economy Excellent chemical stability Resin fulfills industrial demands for security of supply, robust performance, and regulatory support Capto Q XP resin is based on a high-flow agarose base matrix with good pressure-flow properties and wide pore openings to give high available surface area for adsorption of large biomolecules. The Q ligand of the resin is well-established in large-scale purifications. The features of the base matrix, in combination with the conventional Q ligand, make Capto Q XP resin an excellent choice for high-productivity capture and intermediate purification of large biomolecules. 116 Product Quantity Product code Capto Q 25 mL 17531610 Capto Q 100 mL 17531602 Capto Q 1 L 17531603 Capto Q 5 L 17531604 Capto Q 60 L 17531660 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Capto Q ion exchange chromatography resin Designed to meet the demands of modern, large-scale manufacturers, the shorter processing times of Capto Q resin can reduce exposure of the target protein. Features and benefits Strong anion exchanger for high-productivity capture and intermediate purification when high volume throughput is essential High dynamic binding capacity at high flow raises productivity High-volume throughput cuts process times Cost-effective processing with smaller unit operations Resin fulfills industrial demands for security of supply, robust performance, and regulatory support Capto Q resin is designed for capture and intermediate purification of proteins from large feed volumes. It is composed of a rigid, high-flow agarose matrix modified with dextran surface extenders and a strong quaternary ammonium (Q) anion exchanger. The combination of high-volume throughput and high dynamic binding capacity for Capto Q resin raises the productivity in industrial downstream processes. 117 Product Quantity Product code Capto S ImpAct 25 mL 17371701 Capto S ImpAct 100 mL 17371702 Capto S ImpAct 1 L 17371703 Capto S ImpAct 5 L 17371704 Capto S ImpAct 10 L 17371705 Capto S ImpAct 60 L 17371760 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Capto S ImpAct ion exchange chromatography resin With a high binding capacity and small particle size, Capto S ImpAct resin is a good choice for reliable and robust polishing steps in an industrial purification process. High binding capacity > 100 mg mAb/mL resin Efficient aggregate removal at high load of monoclonal antibodies High-resolution polishing based on the well-established Capto resin platform Flexibility of design thanks to a large operational window of flow rates and bed heights for easy optimization and scaling High productivity enabling cost-efficient manufacturing Resin fulfills industrial demands for security of supply, robust performance, and regulatory support This chromatography resin is part of our platform of high-resolution resin based on the Capto resin product line. The polymeric ligand, in combination with the attributes of the base matrix, gives this cation exchanger a high binding capacity. Furthermore, the small bead size enables high-resolution purifications. 118 Product Quantity Product code Capto S 25 mL 17544110 Capto S 100 mL 17544101 Capto S 1 L 17544103 Capto S 5 L 17544104 Capto S 10 L 17544105 Capto S 60 L 17544160 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Capto S ion exchange chromatography resin Capto S resin is a strong cation exchanger for fast, efficient, and cost-effective capture and intermediate protein purification from large feed volumes. Strong cation exchanger for high-productivity capture and intermediate purification when high volume throughput is essential High dynamic binding capacity at high flow raises productivity High-volume throughput cuts process times Cost-effective processing with smaller unit operations Resin fulfills industrial demands for security of supply, robust performance, and regulatory support Capto S resin is designed for capture and intermediate purification of proteins from large feed volumes. It is composed of a rigid, highflow agarose matrix modified with dextran surface extenders and a strong sulfonate (S) cation exchanger. 119 Product Quantity Product code Capto SP ImpRes 25 mL 17546810 Capto SP ImpRes 100 mL 17546802 Capto SP ImpRes 1 L 17546803 Capto SP ImpRes 5 L 17546804 Capto SP ImpRes 10 L 17546805 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Capto SP ImpRes ion exchange chromatography resin With a high binding capacity and small bead size, Capto SP ImpRes strong cation exchange resin is a good choice for reliable and robust polishing steps in industrial purification. Features and benefits High-resolution intermediate purification and polishing resin based on the well-established Capto resin platform with traditional ligands; smaller particle size compared with Capto S Flexible process design due to large operational window of flow rates and bed heights High throughput purifications easy to optimize and scale-up Higher manufacturing productivity enables improved process economy Resin fulfills industrial demands for security of supply, robust performance, and regulatory support Capto SP ImpRes resin is a strong cation exchanger for the highthroughput intermediate purification and polishing steps of a wide range of biomolecules. This chromatography resin is part of an expanded high-resolution platform based on the high-flow agarose Capto resin product line. By combining the high-flow characteristics of Capto resin with a small particle size, Capto SP ImpRes resin delivers excellent pressure-flow properties with impressive resolution. The ability to run at higher flow rates and higher bed heights also increases flexibility in process design. 120 Product Quantity Product code RESOURCE Q columns 1 mL 17117701 RESOURCE Q columns 6 mL 17117901 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection RESOURCE™ Q columns RESOURCE™ Q columns are prepacked with SOURCE™ 15Q resin, a strong anion exchanger for high resolution polishing purification of proteins at high flow rates. Quaternary ammonium (Q) strong anion exchanger for high resolution ion exchange chromatography. Features and benefits Monosized 15 µm rigid beads give low back pressure and high flow rates allowing for high productivity. High chemical stability enables wide range of working conditions with good resistance to cleaning conditions at high pH. Rigid, monodispersed spherical SOURCE 15Q resin beads (15 µm) with controlled pore-size distribution facilitate high-resolution purification at high flow rates. Hydrophilization of the beads further minimizes nonspecific adsorption and allows for high recovery rates for purified samples. The material of the RESOURCE Q column body is polyetheretherketone (PEEK). This ion exchange chromatography column generates low back pressures and high performance, making it excellent for ion exchange (IEX) chromatography on a large scale. 121 Product Quantity Product code RESOURCE RPC 1 mL 17118101 RESOURCE RPC 3 mL 17118201 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection RESOURCE RPC columns RESOURCE RPC columns are prepacked with SOURCE 15RPC resin designed for rapid screening experiments, method development, and small-scale protein purification using reversed phase chromatography. Features and benefits Wide pH range (1 to 12), outstanding selectivity, chemical resistance, high capacity, and high resolution at high flow rates Excellent scalability, from RESOURCE to FineLINE™ columns Using ÄKTA system design and other high-performance liquid chromatography systems SOURCE 15RPC resin is based on rigid, monosized 15 µm diameter polystyrene/divinyl benzene beads. The matrix has outstanding selectivity for RPC. The monodispersity of the beads yields stable beds, low back pressures and excellent results at high flow rates. With its high physical and chemical stability and very high batchto-batch reproducibility, SOURCE 15RPC resin is well suited for all stages of an industrial scale operation — from research and process development through scale-up and into production. SOURCE 15RPC resin offers properties superior to those of other polymeric matrices. 122 Product Quantity Product code RESOURCE S columns 1 mL 17117801 RESOURCE S columns 6 mL 17118001 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection RESOURCE S columns RESOURCE S columns are prepacked with SOURCE 15S resin, a strong cation exchanger for high-resolution polishing purification of proteins at high flow rates. Features and benefits Sulfoethyl (S) strong cation exchanger for high resolution ion exchange chromatography Monosized 15 µm rigid beads give low back pressure and high flow rates allowing for high productivity High chemical stability enables wide range of working conditions with good resistance to cleaning conditions at high pH Rigid, monodispersed spherical SOURCE 15S resin beads (15 µm) with controlled pore-size distribution facilitate high-resolution purification at high flow rates. Hydrophilization of the beads further minimizes nonspecific adsorption and allows for high recovery rates for purified samples. The material of the RESOURCE S column body is PEEK. This ion exchange chromatography column generates low back pressures and high performance, making it excellent for ion exchange (IEX) chromatography on a large scale. 123 Product Quantity Product code RESOURCE ISO 1 mL 17118501 Product Quantity Product code RESOURCE PHE 1 mL 17118601 RESOURCE ISO columns RESOURCE ISO columns are prepacked with SOURCE 15 ISO (isopropyl) resin for purification of proteins and peptides with hydrophobic interaction chromatography (HIC). For fast, high-resolution purifications, RESOURCE columns are excellent choice for polishing and removal of trace impurities. Features and benefits High performance at low back pressure, even with high sample load of up to 25 mg of protein Hydrophobic ligand (isopropyl) coupled to the 15 µm monodispersed rigid polystyrene/divinylbenzene matrix Designed for low- to medium- pressure systems ranging from a peristaltic pump to ÄKTA systems and FPLC systems. RESOURCE HIC columns are available with ether, isopropyl, or phenyl. Generally, RESOURCE PHE will have the strongest hydrophobicity followed by RESOURCE ISO and RESOURCE ETH, successively. RESOURCE PHE columns RESOURCE PHE columns are prepacked with SOURCE 15 PHE (phenyl) resin for purification of proteins and peptides with HIC. For fast, high-resolution purifications, RESOURCE columns are excellent choice for polishing and removal of trace impurities. High performance at low back pressure, even with high sample load of up to 25 mg of protein Hydrophobic ligand (phenyl) coupled to the 15 µm monodispersed rigid polystyrene/divinylbenzene matrix Designed for low- to medium- pressure systems ranging from a peristaltic pump to ÄKTA systems and FPLC systems RESOURCE HIC columns are available with ether, isopropyl, or phenyl. Generally, RESOURCE PHE will have the strongest hydrophobicity followed by RESOURCE ISO and RESOURCE ETH, successively. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 124 Product Quantity Product code Superdex 200 increase 3.2 × 300 mm 28990946 Superdex 200 increase 5 × 150 mm 28990945 Superdex 200 increase 10 × 300 mm 28990944 Gel filtration Superdex™ 200 increase small-scale columns Superdex™ 200 increase size exclusion chromatography (SEC) columns are well suited for high-resolution analysis and small-scale purification of monoclonal antibodies (mAb) and other biomolecules with Mr ~ 10 000 to ~ 600 000. Features and benefits Enhanced performance: improved resolution and runtime compared to predecessor Superdex 200 columns Very high resolution: small bead size and narrow particle size distribution provide high resolution for high protein purity High flow rates: rigid beads give excellent pressure/flow properties “Increase”: refers to size exclusion chromatography columns from Cytiva Superdex 200 increase resin is a next-generation, dextranagarose composite matrix for SEC. With smaller and more rigid beads than their predecessors, Superdex increase columns deliver higher resolution purification in shorter run times. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Superdex 200 increase resin has a selectivity curve optimized for excellent resolution of antibodies and macromolecules within the range Mr ~ 100 000 to ~ 300 000. It is well suited for detection and separation of antibody monomers, dimers, and aggregates present in monoclonal antibody preparations. The outstanding resolving power of this resin facilitates protein characterization, including analysis of antibody size variants, assessment of membrane protein size homogeneity, and the study of protein-protein interactions. Superdex 200 increase resin is optimized to give high resolution separation for proteins with molecular weights in the range Mr ~ 100 000 to ~ 300 000. Superose™ 6 increase resin provides a complementary fractionation range, resolving very large protein complexes and macromolecules that Superdex 200 increase resin cannot separate. 125 Product Quantity Product code Superdex 75 increase 3.2 × 300 mm 29148723 Superdex 75 increase 5 × 150 mm 29148722 Superdex 75 increase 10 × 300 mm 29148721 Higher resolution and/or reduced runtime compared with predecessor, Superdex 75 increase resin belongs to the new generation of agarose-based SEC media, comprising smaller, more rigid beads with a narrower particle size distribution and a higher selectivity for improved performance. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Superdex™ 75 increase columns Superdex™ 75 increase columns are prepacked columns for versatile use in size exclusion chromatography applications. The columns are intended for use in preparative purification as well as for characterization and analysis of proteins with molecular weights between Mr 3000 and 70 000, such as recombinant tagged proteins. Features and benefits Up to 50% higher resolution compared with predecessor Superdex 75 resin, for improved purity and analysis results Runtime reduced down to one third compared with predecessor Superdex 75 resin, with maintained resolution for results faster Versatile use in both preparative and analytical applications, especially with recombinant tagged proteins Higher lot-to-lot consistency compared with predecessor Superdex 75 resin Tolerance to repeated harsh cleaning procedures at high pH ensures long column lifetime and minimizes carryover 126 Product Quantity Product code Superdex 200 prep grade 150 mL 17104301 Superdex 200 prep grade 1 L 17104302 Superdex 200 prep grade 5 L 17104304 Superdex 200 prep grade 10 L 17104305 Superdex 200 prep grade 60 L 17104306 Superdex 200 prep grade resin Superdex 200 prep grade size exclusion chromatography resin offers very high-resolution fractionation for separation, polishing, and product formulation of monoclonal antibodies. Features and benefits Size exclusion chromatography resin providing exceptional resolution with short run times and good recovery Superdex 200 prep grade resin is recommended for the purification of monoclonal antibodies BioProcess resin supported for industrial applications and well-established in approved processes Superdex prep grade resin is a preparative size exclusion chromatography resin with a composite matrix of dextran and agarose. This matrix combines the excellent size exclusion chromatography properties of cross-linked dextran (Sephadex™ resin) with the physical and chemical stabilities of highly cross-linked agarose, to produce a separation resin with outstanding selectivity and high resolution. In addition, its low nonspecific interaction permits high recovery of biological material. Together these properties make Superdex prep grade resin an optimal size exclusion chromatography resin for all applications from laboratory to process scale. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Superdex prep grade resin is available in a range of different bulk pack sizes and convenient prepacked formats for easy scale-up and process development. As a BioProcess resin, Superdex prep grade resin meets industrial demands with security of supply and comprehensive technical and regulatory support. Superdex prep grade resin is available in three selectivities: Superdex 30 prep grade resin is designed for purification of molecules in the molecular weight range up to Mr 10 000, such as peptides, oligosaccharides, and small proteins. Superdex 75 prep grade resin is recommended for the purification of recombinant proteins (fractionation range Mr 3000 to 70 000). Superdex 200 prep grade resin is recommended for the purification of monoclonal antibodies (fractionation range Mr 10 000 to 600 000). 127 Product Quantity Product code Superdex 75 increase 5 × 150 mm 29148722 Superdex 75 increase 3.2 × 300 mm 29148723 Superdex 75 increase 10 × 300 mm 29148721 Superdex 75 increase resin belongs to the new generation of agarose based SEC media, comprising smaller, more rigid beads with a narrower particle size distribution and a higher selectivity for improved performance compared with its predecessor Superdex 75 resin. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Superdex 75 increase columns Superdex 75 increase columns are prepacked columns for versatile use in size exclusion chromatography applications. The columns are intended for use in preparative purification as well as for characterization and analysis of proteins with molecular weights between Mr 3000 and 70 000, such as recombinant tagged proteins. Up to 50% higher resolution compared with predecessor Superdex 75 columns, for improved purity and analysis results. Features and benefits Runtime reduced down to one third compared with predecessor Superdex 75 columns, with maintained resolution for faster results Versatile use in both preparative and analytical applications, especially with recombinant tagged proteins Higher lot-to-lot consistency compared with predecessor Superdex 75 columns Tolerance to repeated harsh cleaning procedures at high pH ensures long column lifetime and minimizes carryover 128 Product Quantity Product code HiLoad Superdex 200 PG 1 × 120 mL 28989335 HiLoad Superdex 200 PG 1 × 120 mL 28989336 HiLoad Superdex 200 preparation grade resin is well suited for rapid, preparative protein purification and scale-up. Its selectivity curve gives a broad fractionation range from Mr ~ 10 000 to 600 000, with excellent resolving power for antibody monomers, dimers, and contaminants of lower molecular weight, such as albumin and transferrin. High mechanical strength and hydrophilicity make this dextranagarose composite resin suitable for larger scale applications that require high flow rates, minimal nonspecific interactions, and fast, effective cleaning-in-place (CIP) protocols. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection HiLoad™ Superdex 200 preparation grade resin columns Designed for high-resolution protein fractionation at preparative scales. Prepacked with Superdex 200 pg resin, these size exclusion chromatography columns enable purification of mAbs, DNA fragments and other biomolecules with Mr ~ 10 000 to ~ 600 000. Features and benefits Preparative-grade: high purity and resolution Two column sizes available: for samples up to 5 mL and up to 13 mL Prepacked columns: convenient and reproducible; can be used with standalone pump or chromatography system Scalable SEC: resin properties and column dimensions optimized for preparative-scale protein purification; choose from two XK column sizes (16/600 or 26/600) Compatible with ÄKTA systems: fittings (1/16”) for direct connection to ÄKTA systems and other chromatography systems 129 Product Quantity Product code HiLoad Superdex 200 PG XK columns 50 × 500 mm 90100040 HiLoad Superdex 200 PG XK columns 50 × 900 mm 90100045 HiLoad Superdex 200 PG XK columns 26 × 900 mm 90100273 HiLoad Superdex 200 PG XK columns 16 × 900 mm 90100137 HiLoad Superdex 200 prep grade columns HiLoad Superdex 200 prep grade columns are prepacked XK columns designed for high-resolution preparative gel filtration chromatography. The column is an excellent choice for separating mAbs from dimers and contaminants. Features and benefits High resolution with short run times and good recovery High capacity at high flow rates Separations can be scaled up to production levels Prepacked for convenience and reproducibility 1/16” fittings for convenient connections to ÄKTA design systems Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Superdex prep grade (pg) resin is produced by covalent bonding of dextran to highly cross-linked agarose. The separation properties of these resins are determined by the dextran component. Steep selectivity curves give excellent resolving power for peptides and proteins in the molecular weight ranges, Mr < 10 000 (Superdex 30 pg resin), Mr 3000 to 70 000 (Superdex 75 pg resin), and Mr 10 000 to 600 000 (Superdex 200 pg resin). The resin combines high mechanical strength with high hydrophilicity, allowing high flow rates, and minimal nonspecific interactions. 130 Product Quantity Product code Superdex 30 150 mL 17090501 Superdex 30 1 L 17090503 Superdex 30 5 L 17090504 Superdex prep grade resin is available in a range of different bulk pack sizes and convenient prepacked formats for easy scale-up and process development. As a BioProcess medium, Superdex prep grade resin meets industrial demands with security of supply and comprehensive technical and regulatory support. Superdex prep grade resin is available in three selectivities: Superdex 30 prep grade resin is designed for purification of molecules in the molecular weight range up to Mr 10 000, such as peptides, oligosaccharides, and small proteins. Superdex 75 prep grade resin is recommended for the purification of recombinant proteins (fractionation range Mr 3000 to 70 000). Superdex 200 prep grade resin is recommended for the purification of monoclonal antibodies (fractionation range Mr 10 000 to 600 000). Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Superdex 30 prep grade gel filtration resin Superdex 30 prep grade gel filtration resin offers very highresolution fractionation for separation, polishing, and product formulation of peptides and small proteins. Features and benefits Gel filtration resins providing exceptional resolution with short run times and good recovery. Superdex 30 prep grade is suitable for polishing and formulation of peptides and small proteins. BioProcess resin supported for industrial applications. Superdex prep grade resin is a preparative gel filtration resin with a composite matrix of dextran and agarose. This matrix combines the excellent gel filtration properties of cross-linked dextran (Sephadex resin) with the physical and chemical stabilities of highly crosslinked agarose, to produce a separation resin with outstanding selectivity and high resolution. In addition, its low nonspecific interaction permits high recovery of biological material. Together, these properties make Superdex prep grade resin an optimal gel filtration resin for all applications from laboratory to process scale. 131 Product Quantity Product code Superdex 30 increase SEC 3.2 × 300 mm 29219758 Superdex 30 increase SEC 10 × 300 mm 29219757 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Superdex 30 increase SEC columns Superdex 30 increase SEC columns are for high resolution, small-scale purification and analysis of small biomolecules with Mr ~ 100 to ~ 7000 such as peptides. Features and benefits Improved performance: improved resolution and runtime compared to predecessor Superdex peptide resin Tolerance to repeated harsh cleaning procedures at high pH ensures long column lifetime and minimizes carryover Very high resolution: small bead size and narrow particle size distribution provide high resolution for high protein purity High flow rate: due to rigid beads giving excellent pressure/flow properties Superdex 30 increase resin is part of a new generation of agarose-based SEC columns from Cytiva called “Increase.” They offer higher resolution purification in shorter runtimes compared with their predecessors. 132 Product Quantity Product code Superose 6 increase HiScale 16 × 40 cm 29321903 Superose 6 increase HiScale 26 × 40 cm 29321904 Superdex 200 increase HiScale 16 × 40 cm 29321905 Superdex 200 increase HiScale 26 × 40 cm 29321906 Superdex 75 increase HiScale 16 × 40 cm 29321907 Superdex 75 increase HiScale 26 × 40 cm 29321908 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection HiScale columns packed with Superdex increase or Superose increase resins If you like our new-generation Superdex increase and Superose increase size exclusion chromatography (SEC) columns but need to purify larger volumes of up to 8.5 mL, we can pack the resin for you in HiScale columns. Features and benefits High-resolution SEC columns: small particle size and narrow particle size distribution of the packed resin ensures highly pure protein HiScale columns packed with SEC resins available on demand, please request a quote HiScale columns are packed on demand for you with Superdex 200 increase, Superdex 75 increase, and Superose 6 increase SEC (gel filtration) resins. The columns are available in two sizes: 16 × 400 mm and 26 × 400 mm. These two column sizes enable purification of sample volumes up to 3.2 mL and 8.5 mL, respectively. 133 Product Quantity Product code HiLoad Superdex 75 PG 16 × 600 mm 28989333 HiLoad Superdex 75 PG 26 × 600 mm 28989334 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection HiLoad Superdex 75 pg prepacked columns HiLoad Superdex 75 pg prepacked columns are for high-resolution size exclusion chromatography of recombinant proteins from samples up to 5 mL or 13 mL. Prepacked format: for convenience and reproducibility; can be used with standalone pump or chromatography system High resolution: for high purity Scalable: separations can be scaled up using Superdex 75 prep grade resin ÄKTA system compatible: fittings (1/16”) for direct connection to ÄKTA systems and other chromatography systems This column is packed with Superdex 75 prep grade (pg) resin. This SEC resin combines excellent mechanical strength with high hydrophilicity, allowing high flow rates and minimal nonspecific interactions. Steep selectivity curves give excellent resolving power for proteins and other biomolecules in the molecular weight range: Mr ~ 3000 to 70 000. 134 Product Quantity Product code HiLoad Superdex 75 PG XK columns 16 × 40 cm 29321903 HiLoad Superdex 75 PG XK columns 26 × 40 cm 29321904 HiLoad Superdex 75 PG XK columns 16 × 40 cm 29321905 HiLoad Superdex 75 PG XK columns 26 × 40 cm 29321906 HiLoad Superdex 75 PG XK columns 26 × 40 cm 29321908 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection HiLoad Superdex 75 prep grade XK columns HiLoad Superdex 75 prep grade columns are prepacked 20 mL XK columns designed for high-resolution, preparative size exclusion chromatography (gel filtration) separating proteins and polynucleotides. Features and benefits High resolution with short run times and good recovery High capacity at high flow rates Separations can be scaled up to production levels Prepacked for convenience and reproducibility 1/16” fittings for convenient connections to ÄKTA chromatography systems Superdex prep grade (pg) resin is produced by covalent bonding of dextran to highly cross-linked agarose. The separation properties of these resins are determined by the dextran component. Steep selectivity curves give excellent resolving power for peptides and proteins in the molecular weight ranges, Mr < 10 000 (Superdex 30 pg resin), Mr 3000 to 70 000 (Superdex 75 pg resin), and Mr 10 000 to 600 000 (Superdex 200 pg resin). The resin combines high mechanical strength with high hydrophilicity, allowing high flow rates, and minimal nonspecific interactions. 135 Product Quantity Product code HiLoad Superdex 30 PG XK columns 16 mm 28989331 HiLoad Superdex 30 PG XK columns 26 mm 28989332 HiLoad Superdex 30 prep grade XK columns HiLoad Superdex 30 prep grade columns are prepacked XK columns designed for high-resolution, preparative gel filtration chromatography separating peptides. Features and benefits High-resolution separation: with short run times and good recovery High capacity: at high flow rates Prepacked: for convenient, ready-to-go use and reproducible results Scalable protein separation: easily scale up separations to production levels Integrated workflows: 1/16” fittings for convenient connection with ÄKTA protein purification systems Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Size exclusion chromatography (SEC) column technologies have played a critical role in biomolecule separation for decades. As protein purification is a common method in labs worldwide, scientists increasingly need reliable and convenient options that meet modern requirements for efficient, high-resolution protein separation. HiLoad Superdex pg SEC columns offer the combination of prepacked convenience and reproducibility together with fast, high-resolution gel filtration at preparative laboratory scale. 136 Product Quantity Product code Superose 6 Increase SEC columns 3.2 × 300 mm 29091598 Superose 6 Increase SEC columns 5 × 150 mm 29091597 Superose 6 Increase SEC columns 10 × 300 mm 29091596 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Superose 6 increase SEC columns Superose 6 increase SEC columns are designed for high-resolution, small-scale preparative purification and analysis of large globular proteins, protein complexes, and other macromolecules in the molecular weight range Mr ~ 5000 to ~ 5 000 000. Features and benefits Wide fractionation range: optimized to purify large biomolecules, including membrane proteins, protein complexes, and DNA fragments Very high resolution: small bead size and a narrow particle size distribution provide high resolution for high protein purity High flow rates: rigid beads give excellent pressure/flow properties Enhanced performance: improved resolution and runtime compared to predecessor Superose 6 increase resin is a next-generation, agarose-based matrix for size exclusion chromatography. With smaller and more rigid beads than their predecessors, Superose 6 increase columns deliver higher resolution protein purification in shorter runtimes. 137 Product Quantity Product code Superose 6 Prep Grade 125 mL 17048901 Superose 6 Prep Grade 1 L 17048903 Superose 6 Prep Grade 5 L 17048904 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Superose 6 prep grade resin Good resolution separations of proteins and other biomolecules according to size, Wide separation range with separations in a molecular weight range from Mr 5000 up to Mr 5 000 000. Features and benefits Excellent reproducibility and durability Achieve high-resolution separations across exceptionally broad molecular-weight ranges Low ionic strength eluents (< 0.05 M) can utilize a hydrophobic interaction component to alter the selectivity of Superose resin for some lipids, peptides, and small aromatic compounds High performance gel filtration resin specially designed for preparative purification of biomolecules Superose 6 resin is designed for high-resolution chromatography. Superose 6 prep grade resin permits easy, efficient scale-up to preparative separations. 138 Product Quantity Product code HiLoad 16/600 Superose 6 prep grade columns 16 × 600 mm 29323952 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection HiLoad 16/600 Superose 6 prep grade columns HiLoad 16/600 Superose 6 prep grade (pg) columns are prepacked size exclusion chromatography (SEC) columns for high-resolution purification of large biomolecules and protein complexes. The column is an excellent choice for purification of sample volumes up to 5 mL. Features and benefits Convenience and reproducibility: can be used with standalone pump or a chromatography system High-resolution separations: purification is possible across an exceptionally broad molecular-weight range Scalable: separations can be scaled up using Superose 6 prep grade resin ÄKTA system compatible: fittings (1/16”) for direct connection to ÄKTA systems and other chromatography systems This column is packed with Superose 6 pg resin, which provides good resolution separations of proteins and other biomolecules according to size. The wide fractionation range of the resin enables reproducible and robust separations in the molecular weight range from Mr 5000 up to Mr 5 000 000. 139 Product Quantity Product code XK columns packed with Superose 6 prep grade 26 × 70 mm 90100043 XK columns packed with Superose 6 prep grade 50 × 60 mm 28923760 Product Quantity Product code Superose 12 PG 125 mL 17053601 Superose 12 PG 1 L 17053603 Superose 12 PG 5 L 17053604 XK columns packed with Superose 6 prep grade resin XK columns packed on-demand. Superose 12 prep grade resin Good resolution separations of proteins and other biomolecules according to size. Features and benefits Wide separation range: separations in a molecular weight range from Mr 1000 up to Mr 300 000 Excellent reproducibility and durability Superose 12 resin is for high resolution purification of proteins from Mr 1000 up to Mr 300 000, while Superose 12 prep grade resin is useful for preparative purification of these higher molecular weight proteins. Custom deep well plates with resin available We apply our expertise in membranes, resins, and mold tooling to develop custom plates for a range of applications and sample throughput levels. Our proprietary process to encapsulate the filter media minimizes contamination between wells. The plates are optimized for automated filling, allowing for greater efficiency. Our dedicated coating facilities are environmentally controlled, meeting strict quality standards. Plates can be tailored to your specifications in terms of labeling and quantity per case. Learn more Learn more about multiwell plates and microplates. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 140 Membrane family selection Membrane surfactants and protein binding: good news and bad news Every nitrocellulose membrane on the market contains a surfactant, usually an anionic surfactant. The exact nature of the surfactant is proprietary for every membrane manufacturer, but the surfactant functions are always the same. First, it makes the membrane hydrophilic. Second, and perhaps even more important, it helps proteins bind to the membrane (Fig 1). The good news: The surfactant partially denatures proteins dispensed onto the membrane in a buffer and helps them bind to nitrocellulose (NC) fibers. The bad news: The surfactant denatures proteins. Denaturing can potentially destroy the antigen binding sites of some monoclonal antibodies completely, making these reagents unusable in a lateral-flow test. From our experience, about 2%–3% of clones are affected by this problem, and others are partially impaired by this process. In these cases, it makes sense to evaluate different membranes with different surfactants for their performance with your specific molecule and reagents. Cytiva offers different families of NC membrane, each containing a specific surfactant at a defined concentration, which is the same across all membrane grades within the family. A summary of the properties of the different Cytiva membranes can be found at the end of this article. Fig 1. Schematic representation of the process of protein binding to an NC membrane. Protein in bulk solution Transport to interfacial region Rearrangement on surface Adsorption and surfactant interation NC fiber Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 141 Binding kinetics of capture reagents and membrane compatibility Whether a specific capture reagent (e.g., a monoclonal antibody) provides the sensitivity and specificity required in a lateral-flow rapid test depends not only on the membrane and the surfactant incorporated in the membrane; it also depends on the kinetic properties of the capture reagents. The affinity of an antibody is determined by its on-rate (the rate at which an antigen binding site grabs its target) and its off-rate (the rate at which the bound antigen is released again from the binding site). Antibodies with identical affinities for a specific target may have very different on- and off-rates, as described in Figure 2. The residence time of an antigen at the test line of a lateralflow test is usually very short — always less than a minute and sometimes only a few seconds. Therefore, antibodies with high on-rates are required for lateral-flow rapid tests. A test developer should evaluate all available capture proteins for their target antigen in the test system itself, or, more favorably, by surface plasmon resonance (SPR) prior to antibody selection. The latter approach enables the developer to exclude clones with obviously unfavorable binding or releasing kinetic properties. Fig 2. Kinetic properties of different monoclonal antibodies with identical affinities for a specific antigen. The data were obtained using surface plasmon resonance with Cytiva Biacore equipment. Antibodies with slow on- and off- rates (left) can easily have the same affinity (center) as antibodies with fast on- and off- rates (right). If a test must be developed with a specific pair of capture and detector reagents simply because there are no alternatives available, there might be a conflict between the sensitivity goals and the goals for test duration that have been defined before the development was started. Capture reagents with slow on-rates may result in a low sensitivity test on a fast membrane, although only this membrane provides the developer with the “right” test duration time. If this happens, it sometimes helps to look for membranes with different surfactants and/or surfactant concentrations. But the ultimate outcome might be that either the original goals of the test development must be adjusted to what is achievable with this set of reagents or new reagents may have to be developed. Fast on / fast off Slow on / slow off Affinity isotherms 2 molecules with identical affinity but very different kinetics Stability Recognition log k0 log kd Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 142 Features and benefits Superb data quality while increasing your throughput eight-fold, compared to one-needle SPR systems Interaction analysis for screening, kinetics, affinity, epitope binning, concentration, and relative potency Higher operational efficiency with capacity options and streamlined assay development in parallel Modular configuration with option for analysis in a GxP regulated environment Rapid optimization of assay conditions and troubleshooting Product Quantity Product code Biacore 8K instrument kit 1 29722782 Biacore 8K+ instrument 1 29283382 Biacore 8K+ instrument and kit 1 29722783 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Biacore™ 8 series (Biacore 8K and Biacore 8K+) Biacore™ 8 series efficiently delivers quality binding data to meet your toughest challenges in screening, characterization, process optimization, and quality control. Our 16 flow-cell surface plasmon resonance (SPR) systems, Biacore 8 series, come in two configurations: Biacore 8K for high-throughput screening and characterization Biacore 8K+ for a significantly higher capacity These eight-needle, high-sensitivity SPR systems rapidly provide reproducible kinetics, affinity and concentration data, and shortens your time to results by up to eight times compared to one needle systems. The blend of system flexibility and throughput reduces the experimental cycle time, even for complex targets and drug formats such as bispecific antibodies. This reduced cycle time offers more opportunity for screening in drug discovery and development of small molecules to large viruses in pure or complex samples. Application methods are easily transferred to other labs or other Biacore 8 series systems and to our one-needle SPR platform Biacore 1 series. 143 Biacore 1 series (Biacore 1K, Biacore 1K+, Biacore 1S+) Biacore 1 series is a one-needle platform that transforms protein interaction analysis using SPR — making it simpler and faster to use without compromising quality. Now you can focus on generating the consistent and reproducible data needed in your research. Our six flow-cell SPR system, Biacore 1 series comes in three system configurations: Biacore 1K, Biacore 1K+, and Biacore 1S+. The systems offer analytical flexibility that can grow with your group’s need for sample capacity, sensitivity, and throughput. All systems are built for analysis in a GxP-regulated environment. The single software platform for Biacore 1 series systems allow you to spend less time training on running the instruments and less time on result generation and evaluation. Application methods are easily transferred to other labs or to other Biacore 1 series systems and the higher-throughput systems of the Biacore 8 series. You can use Biacore 1 series systems across a wide range of applications, molecules, and both pure and complex samples — from small fragments to large viruses. The systems are scalable, so your research won’t be limited as your needs evolve. Product Quantity Product code Biacore 1K SPR system 1 29726017 Biacore 1K+ SPR system 1 29726018 Biacore 1S+ SPR system 1 29726019 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Read more Read our Biacore systems selection guide. Read more Read our Biacore consumables selection guide. Download Download our poster to save time and effort for your Biacore SPR interaction analysis study. Features and benefits No programming skills needed for setup and to start analysis when using predefined methods and flexible software tools to speed up assay development Straightforward transfer of methods to other Biacore 1 series or Biacore 8 series systems Less time needed to train on running the instrument and how to generate and evaluate results Simpler data interpretation: compile, visualizev and export data with results in minutes Optimized injection design with six flow cells enables more efficient sample utilization and greater application utility, including multicomplex analyses in one run Maximize run efficiency by queuing methods and assays and let it run overnight or over the weekend 144 Series S sensor chip CM5 Biacore sensor chip CM5 supports a range of immobilization chemistries to capture nucleic acids, peptides, antibodies, proteins, carbohydrates, or chemical compounds for interaction analysis from research to discovery, development, and quality control. The CM5 sensor chip carries a matrix of carboxymethylated dextran covalently attached to a gold surface. Molecules can be covalently coupled to the sensor surface by exploiting available amine, thiol, aldehyde, or carboxyl functional groups on the ligand. The CM5 dextran matrix extends about 100 nm from the gold surface and is flexible to allow relatively free movement of attached ligands. Sensor chip CM5 is the most versatile and widely used dextranbased chip, suitable for most general-purpose surface plasmon resonance (SPR) applications. Carboxymethyl surface chemistry supports the study of many different types of ligands and binding partners from small organic molecules, such as drug candidates, through to large molecular assemblies and even whole viruses. High binding capacity gives a high response, which is advantageous for capture assays and for interactions involving small molecules. High surface stability provides accuracy and precision and allows repeated analysis on the same surface. Product Quantity Product code Series S sensor chip CM5 Pack of 10 29149603 Series S sensor chip CM5 Pack of 3 BR100530 Series S sensor chip CM5 Pack of 1 29104988 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Application examples include: FBDD-based screening for novel drugs against Hepatitis C virus (HCV) Surface plasmon resonance-based determination of adenovirus concentration on Biacore T200 Features and benefits First choice for immobilization via -NH2, -SH, -CHO, -OH, or -COOH groups Attach proteins, antibodies, nucleic acids, carbohydrates, and small molecules (< 1000 Da) Compatible with a wide range of applications from basic research to quality control Suitable for ligand fishing and fragment-based drug discovery (FBDD) High immobilization capacity permits a broad range of capture densities Obtain real-time data on binding specificity, affinity, kinetics, concentration and more 145 Product Quantity Product code Series S sensor chip CM3 Pack of 1 29104990 Series S sensor chip CM3 Pack of 3 BR100536 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Series S sensor chip CM3 Biacore sensor chip with similar properties to sensor chip CM5, suited to large interaction partners and exploratory assay conditions. The shorter dextran chains of the CM3 allows the interaction to take place closer to the surface, which can improve sensitivity when working with large molecules, molecular complexes, viruses, or whole cells. Features and benefits Shorter dextran matrix and similar charge density to explore alternative assay conditions Use when the interaction partner in solution is very large Improves sensitivity when monitoring certain interactions Maintains sensitivity by keeping large particles as close as possible to the surface Attach proteins, nucleic acids, carbohydrates, or small molecules Couple to carboxyl groups on the sensor surface via -NH2, -SH, -CHO, -OH or -COOH. 146 Series S sensor chip protein A Biacore sensor chip for orientation-specific binding of antibodies (predominantly human) through Fc-region. The surface is a carboxymethylated dextran matrix with a recombinant protein A variant covalently attached. The surface ligand is the same as is used for our MabSelect SuRe™ protein purification products. Binds antibodies from several mammalian species, most notably human antibodies of the subclasses IgG1, IgG2, and IgG4 Binds only to the heavy chain within the Fc region, ensuring a specific orientation of the antibody on the surface Does not bind Fab fragments (unlike native protein A) Sensor chip protein A is a sensor surface pre-immobilized with a recombinant protein A variant binding antibodies for protein interaction analysis using Biacore systems. It is ready-to-use with high binding capacity offering a wide dynamic range and high convenience, reproducibility, and robustness. Sensor chip protein A is an excellent surface choice for antibody quantitation and characterization in biopharmaceutical, late-stage development and manufacturing QC. Product Quantity Product code Series S sensor chip protein A Pack of 3 29127556 Series S sensor chip protein A Pack of 1 29127555 Features and benefits MabSelect SuRe ligand on the surface Site-directed capture of antibody: binds predominantly to the heavy chain within the Fc region and ensures antibodies are bound to the surface in a specific orientation Time and effort savings: ready-to-use sensor chip eliminates the need to develop immobilization and regeneration conditions Confidence in data: highly controlled development processes and manufacturing conditions ensure reproducible results Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 147 Features and benefits Combines convenience with high reproducibility and robustness in the analysis of interactions where antibodies are used as ligands Binds only to the heavy chain within the Fc region ensuring antibodies are bound to the surface in a specific orientation Ready-to-use, eliminating the need to develop immobilization and regeneration conditions, saving time and effort Product Quantity Product code Series S sensor chip protein G Pack of 1 29179315 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Series S sensor chip protein G A Biacore Extend product used for oriented capture or binding of a wide range of mammalian and all human antibody subclasses. The surface is a carboxymethylated dextran matrix pre-immobilized with a recombinant protein G — GammaBind™ G, type 2. The recombinant protein G binds a broad range of IgG, such as human (including IgG3), rat, rabbit, mouse, guinea pig, goat, sheep, and cow. Sensor chip protein G is one of an expanding range of Biacore Extend products. These sensor chips provide researchers working in nonregulated environments with the convenience of a wider choice of consumables optimized for Biacore applications. What to expect from Biacore Extend product: Ordered as standard products Same customer support system is used as for a regular product QC performed, but no Certificate of Analysis/Conformance issued For some products, lot-to-lot variation might be higher compared to a standard product 148 Product Quantity Product code Series S sensor chip protein L Pack of 1 29205138 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Series S sensor chip protein L A Biacore Extend sensor chip for site-directed antibody capture. Suitable for study of a wide range of antibody fragments including Fabs and dAbs. The surface is a carboxymethylated dextran matrix pre-immobilized with a recombinant protein L for antibodies and antibody fragments containing kappa light chain subtypes (1, 3, and 4) without interfering with its antigen-binding site. Protein L is suitable for the capture of a wider range of antibody classes than both protein A and protein G, including IgG, IgM, IgA, IgE, and IgD. Protein L does not bind bovine immunoglobulins, which often contaminate serum supplements, and is therefore especially useful for quantifying antibodies and antibody fragments from serum-based culture. Features and benefits Use for oriented capture of antibody fragments Ready-to-use, eliminating the need to develop immobilization and regeneration conditions, saving time and effort Captures a wide range of antibody fragments, such as Fabs; single-chain variable fragments (scFv); kappa light chain subtypes 1, 3, and 4; and domain antibodies (dAbs 149 Series S sensor chip SA This high-affinity streptavidin (SA) sensor chip immobilizes biotinylated molecules for surface plasmon resonance (SPR) interaction analysis on Biacore systems. Sensor Chip SA provides a surface of streptavidin covalently attached to a carboxymethyl dextran matrix. Dissociation of biotinylated ligands from the surface of the chip is generally negligible during a Biacore analysis because the affinity of streptavidin for biotin is extremely high, with an equilibrium dissociation constant of about 10 to 15 M. High binding capacity, reproducibility, and chemical resistance give excellent performance over a broad range of applications. The surface of sensor chip SA is resistant to 1-minute pulses of many agents commonly used in SPR sensor chip washing and regeneration protocols. Biotin has become a popular molecular tag for capture and interaction studies because it is stable and rarely interferes with target activity or structure due to its small size. Biotin can be conveniently conjugated to a wide range of target molecules by chemical or enzymatic means. Applications for sensor chip SA are therefore broad. Examples include: characterizing PROTAC ternary complex formation screening of endocrine-disrupting chemicals (3) elucidating DNA- and RNA- mediated mechanisms of catechins in cancer prevention (4) Features and benefits Versatile: immobilizes biotinylated peptides, proteins, nucleic acids, or carbohydrates Ready-to-use: streptavidin is pre-immobilized, ready for fast, high-affinity ligand capture Robust: streptavidin-biotin is one of the strongest noncovalent interactions in nature Noncovalent: a convenient alternative for ligands refractory to covalent immobilization Standardized ligand orientation: controlled biotinylation enables oriented capture Product Quantity Product code Series S sensor chip SA Pack of 3 BR100531 Series S sensor chip SA Pack of 1 29104992 Series S sensor chip SA Pack of 10 29699621 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 150 Series S sensor chip NTA Biacore sensor chip NTA captures histidine-tagged molecules by metal chelation for subsequent surface plasmon resonance (SPR) interaction analysis. Histidine is the most widely used molecular tag today. Sensor Chip NTA can be used to capture many different types of polyhistidinetagged proteins for subsequent SPR assays. Interaction of immobilized proteins with a wide range of analyte molecules can be studied, from low-molecular-weight compounds to large proteins. For experiments where low-molecular-weight analytes are studied, sensor chip NTA is the first choice. Sensor chip NTA consists of carboxymethylated dextran with covalently immobilized nitrilotriacetic acid (NTA). The NTA molecule chelates metal ions such as nickel (Ni2+), creating coordination sites that bind to polyhistidine tags on recombinant proteins and other biomolecules. Noncovalent capture of ligands, for example by metal ion chelation, has a number of advantages over covalent immobilization. Tagged proteins can be captured directly from crude cell extracts or culture medium, requiring little or no sample preparation. Because fresh ligand is captured for each cycle, the same chip surface can be used for analysis interactions involving different ligands. Regeneration conditions are generic and therefore the need for assay development is minimized. In addition, capture generates a directed structural orientation of the protein on the surface, potentially offering optimal site exposure. Product Quantity Product code Series S sensor chip NTA, pack of 3 Pack of 3 BR100532 Series S sensor chip NTA, pack of 1 Pack of 1 28994951 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Features and benefits Versatile: captures many types of histidine-tagged molecules Ready-to-use: NTA is pre-immobilized, ready for nickel loading and ligand capture Convenient: compatible NTA reagent kit provides nickel ion and regeneration solutions Easily regenerated: EDTA injection efficiently removes metal ions to regenerate the sensor Chelation capture: simpler assay development, potential to standardize target orientation 151 His capture kit type 2 His capture kit, type 2 is designed for capture of histidine-tagged proteins to an anti-histidine antibody and provides an alternative to capture on nickel-chelated nitrilotriacetic (NTA) groups. The anti-histidine antibody is a monoclonal antibody directed against polyhistidine tags in the C- or N-terminus of a protein. Depending on the nature of the interacting molecules and the micro-environment around the histidine tag, the affinity of the capture antibodies will vary. Features and benefits Enables capture of histidine-tagged ligands; physiological conditions can be used during the coupling Optimized and verified assay minimizing assay development Protocol and required solutions provided Product Quantity Product code His capture kit, type 2 1 29234602 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 152 Product Quantity Product code Biotin CAPture kit, series S 1 28920234 Biotin CAPture kit series S Kit containing reagents and sensor chip for reversible capture of biotinylated molecules. Series S sensor chip CAP Biotin CAPture reagent Regeneration stock 1 Regeneration stock 2 The ligand is captured on to series S sensor chip CAP via Biotin CAPture reagent, which is a modified streptavidin. Series S sensor chip CAP should only be used together with biotin CAPture reagent. Regeneration of the surface after each analysis cycle removes biotin CAPture reagent as well as the ligand and any bound analyte. Fresh biotin CAPture reagent is attached to the surface for each cycle. Sensor ship CAP and biotin CAPture reagent contain deoxyribo-oligonucleotides. Therefore, the kit is not suitable for work with DNA-binding proteins or enzymes that degrade DNA. The ligand capture capacity of sensor chip CAP and biotin CAPture reagent is typically about 1500 to 3000 RU for a ligand with Mr 150 000. Biotin CAPture kit has been optimized for an assay temperature of 25°C. Download Download our poster on Reversible capture of biotinylated molecules for Biacore analysis using Biotin CAPture kit. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Features and benefits Study interactions without the need to know or establish immobilization methods or regeneration protocols; only biotinylation of the ligand is required Study unstable ligands or ligands for which no regeneration can be established Maximize economy and efficiency when working with multiple ligands For most types of interaction analyses in Biacore systems Sufficient for 100 regenerations 153 Gluthatione-S transferase (GST) capture kit Reagents for site-directed affinity capture of GST fusion proteins in biomolecular interaction analyses using Biacore systems. Goat anti-GST antibody, 0.8 mg/mL in 75 µL coupling solution, 5 mL positive control: recombinant GST (Schistosoma japonicum), 0.2 mg/mL in 100 µL HBS-EP regeneration solution, 70 mL. The use of these products in Biacore systems requires an amine coupling kit with sensor chip CM5, CM4, CM3, or C. Not for use with Biacore Flexchip Anti-GST antibody is suitable for immobilization on carboxyl-derivatized sensor chips using amine coupling kit and the included immobilization buffer. Regeneration solution is used for regeneration of the surface by removal of the captured fusion protein. Recombinant GST is used in preparation of the surface and may also be used in assay development work to check that the sample does not bind to GST. Features and benefits For most types of interaction analyses in Biacore systems Sufficient for 20 immobilizations and up to 600 regenerations Product Quantity Product code GST capture kit 1 BR100223 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 154 Human antibody capture kit type 2 Human antibody capture kit, type 2 is intended for capture of human or humanized IgG antibodies as ligands in biomolecular interaction analyses. Anti-human IgG (Fc) antibody is a monoclonal mouse antihuman IgG (Fc) antibody of IgG1 subclass, prepared by affinity chromatography on protein A. The antibody recognizes an epitope within the CH₂ domain in human IgG Fc of all subclasses. It is also known to bind to monkey (cynomolgus) and rabbit IgG. Features and benefits Binds an epitope in CH₂ domain of human IgG (all subclasses recognized) Little or no cross-reactivity towards other human Ig isotypes No, or very low, cross-reactivity with IgG from other species, except other primates and rabbit Very stable binding — little or no dissociation of captured antibodies Product Quantity Product code Human antibody capture kit, type 2 1 29234600 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 155 Product Quantity Product code Human Fab capture kit, type 2 1 29234601 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Human Fab capture kit type 2 Human Fab binder is a mixture of two monoclonal antibodies recognizing, respectively, kappa and lambda subtypes of human Fab fragment light chains. It binds specifically to the Fab region of human antibodies and no binding to the Fc region has been observed. It recognizes most subtypes of both kappa and lambda light chains and does not cross-react with Fab from other species. In addition, no nonspecific binding from cell lysate or cell culture medium has been observed. The main applications of human Fab capture kit, type 2 are screening and characterization of human Fab fragments generated from phage display (purified or directly from crude cell lysates or periplasmic extract). Features and benefits Early selection of binders enabled through kinetics screening by high resolution off-rate ranking Broad framework specificity; kappa and lambda types of human Fab fragments captured Excellent assay performance; fab binder with high capture efficiency and stability 156 Mouse antibody capture kit type 2 Mouse antibody capture kit, type 2 is intended for capture of mouse antibodies as ligands in various biomolecular interaction analyses. Anti-mouse antibodies are suitable for immobilization on carboxyl derivatized surfaces using amine coupling kit and the included immobilization buffer. The regeneration solution is used for regeneration of the surface. Anti-mouse antibodies consist of polyclonal rabbit anti-mouse immunoglobulin antibodies reacting with all IgG subclasses (IgG1, IgG2a, IgG2b, and IgG3), IgA and IgM. Reaction with other mouse immunoglobulin classes has not been investigated, but the antibodies are likely to react with these via their light chains. Features and benefits Binds all IgG subclasses (IgG1, IgG2a, IgG2b, and IgG3), IgA, and IgM. Very stable binding with no or very low dissociation of captured antibodies. Product Quantity Product code Mouse antibody capture kit, type 2 1 29215281 Read more Read more about protein purification methods: ‘Your guide to protein purification basics, tips, and tricks, and more’. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 157 Sample collection Sample is collected and may need to be transferred to liquid media to allow application. Bespoke sample preparation support can be provided upon request. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 158 Chromatography Cellulose fiber pads The base cellulose is a key part of the system, and the correct choice of absorbency, wicking rate, and wet strength are critical to producing a working assay. The Cytiva range of cellulose materials for dipstick colorimetric assays offers highly consistent and inert substrates for absorption of the active chemicals required for development of dipstick tests. CF2 CF-2 is a wet-strengthened, cellulose fiber for use in dipstick colorimetric assays that contains an FDA-approved resin that binds the cellulose fibers together. Features and benefits High strength of the paper, especially when it is wet, reduces waste during manufacture due to the snapping of the cellulose during coating Reduced risk of the pad decomposing during use. Easier handling during kit manufacture. CF3 CF3 is a medium weight, 100% cotton linter material. Consistent absorbency and wicking rates for test-to-test reproducibility. Product manufactured in controlled environments from highquality materials minimizes the risk of false results due to sample contamination. Low protein binding for high test sensitivity and minimal loss of analyte. Naturally hydrophilic for rapid rewetting after prolonged storage. Compatible with most housing styles. CF1 CF1 is a light and thin, 100% cotton linter material (176 µm thickness at 53 kPA), suitable for use as a sample pad for small volume lateral-flow applications. Features and benefits Light, thin grade suitable for small volume. Consistent absorbency and wicking rates for test-to-test reproducibility. Product manufactured in controlled environments from highquality materials minimizes the risk of false results due to sample contamination. Low protein binding for high test sensitivity and minimal loss of analyte. Naturally hydrophilic for rapid rewetting after prolonged storage. Compatible with most housing styles. Minimal leakage along the strip to minimize the risk of contaminating test results. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 159 CF4 CF4 is a medium weight, 100% cotton linter material. Features and benefits Consistent absorbency and wicking rates for test-to-test reproducibility. Product manufactured in controlled environments from high-quality materials minimizes the risk of false results due to sample contamination. Low protein binding for high test sensitivity and minimal loss of analyte. Naturally hydrophilic for rapid rewetting after prolonged storage. CF5 CF5 is a medium to thick weight, 100% cotton linter material. Features and benefits Consistent absorbency and wicking rates for test-to-test reproducibility. Product manufactured in controlled environments from high-quality materials minimizes the risk of false results due to sample contamination. Low protein binding for high sensitivity and minimal loss of analyte. Naturally hydrophilic for rapid rewetting after prolonged storage. Compatible with most housing styles. Minimal leakage along the strip to minimizes the risk of contaminating test results. CF6 CF6 is a thick material suitable for high sample volume applications. Features and benefits Consistent absorbency and wicking rates for test-to-test reproducibility. Product manufactured in controlled environments from high-quality materials minimizes the risk of false results due to sample contamination. Naturally hydrophilic for rapid rewetting after prolonged storage. Compatible with most housing styles. CF7 CF7 is a thick, 100% cotton linter, thick material, suitable for high sample volume applications. Features and benefits Consistent absorbency and wicking rates for test-to-test reproducibility. Product manufactured in controlled environments from high-quality materials minimizes the risk of false results due to sample contamination. Low protein binding for high test sensitivity and minimal loss of analyte. Naturally hydrophilic for rapid rewetting after prolonged storage. Compatible with most housing styles. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 160 Grade Thickness (μm @ 53 kPA) Water absorption (mg/cm2) Dimensions Product code CF 1 176 18.7 22 mm × 50 m 8111-2250 CF 2 172 16.1 22 mm × 50 m 8112-2250 CF 3 322 34.6 22 mm × 50 m 8113-2250 CF 4 782 49.9 22 mm × 50 m 8114-2250 CF 4 782 49.9 210 × 293 mm 8114-6621 CF 5 954 63.3 22 mm × 50 m 8115-2250 CF 6 1450 136.3 22 mm × 50 m 8116-2250 CF 6 1450 136.3 20 mm × 50 m 8116-613 CF 6 1450 136.3 25 mm × 100 m 8116-7007 CF 6 1450 136.3 210 × 293 mm 8116-6621 CF 6 1450 136.3 25 mm × 50 m 8116-9915 CF 7 1873 252.3 22 mm × 50 m 8117-2250 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 161 Grade Thickness (μm @ 53 kPA) Water absorption (mg/cm2) Dimensions Product code GR470 840 78 µm 14 mm × 100 m 9890-10010 GR470 840 78 µm 25 mm × 100 m 9890-10056 GR470 840 78 µm 26.5 mm × 100 m 9890-10195 GR470 840 78 µm 25 mm × 150 m 1053-10020 GR470 840 78 µm 31 mm × 500 ft 9890-10047 GR470 840 78 µm 20 mm × 100 m 9890-10077 GR470 840 78 µm 20 mm × 285 mm 9890-10083 GR470 840 78 µm 18 mm × 50 m 10534267 GR470 840 78 µm 4” × 250 ft 10534408 GR470 840 78 µm 17 mm × 300 mm 10534807 GR470 840 78 µm 26 mm × 100 m 10535012 GR470 840 78 µm 4” × 312 ft 10535174 GR470 840 78 µm 18 × 285 mm 10537731 Table continued > Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection GR470 A thick, absorbent, smooth-surfaced, cotton linter paper for medium and high sample volumes and flow rates. Untreated bound cellulose suitable for whole blood or serum. Originally used as a filter for gelatinous precipitates and oils. Features and benefits Performs well with one or two drops of whole blood Extensively used in clinical labs as an absorbent pad for culture media or as absorbent sink paper 162 Grade Thickness (μm @ 53 kPA) Water absorption (mg/cm2) Dimensions Product code GR470 840 78 µm 19 × 285 mm 10539031 GR470 840 78 µm 10 × 12” 10539036 GR470 840 78 µm 29 × 275 mm 10539044 GR470 840 78 µm 6” × 500 ft 10539067 GR470 840 78 µm 6.5” × 1000 ft 10539068 GR470 840 78 µm 1” × 350 ft 10539554 GR470 840 78 µm 20 mm × 500 ft 10539558 GR470 840 78 µm 18 mm × 100 m 10539998 GR470 840 78 µm 31 mm × 50 m 989010043 GR470 840 78 µm 19 mm × 100 m 989010098 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 163 Whatman glass fiber pads Standard 14 Standard 14 is a glass fiber (355 µm thickness at 53 kPA) suitable for use as a conjugate release pad in a lateral-flow assay. Features and benefits Light, thin glass fiber: suitable for small volumes Consistent absorbency and wicking rates: ensures test-totest reproducibility Product manufactured in controlled environments from high-quality materials: minimizes the risk of false results due to sample contamination Low protein binding: high test sensitivity with minimal loss of analyte Naturally hydrophilic: rapid rewetting after prolonged storage Compatible: works with most housing styles Minimal leakage along the strip: minimizes the risk of contaminating test results Fast flowing: faster flow than cotton with lower sample retention Standard 17 Standard 17 is a glass fiber (370 µm thickness at 53 kPA) suitable for use as a conjugate release pad in a lateral-flow assay. Features and benefits Light, thin glass fiber: suitable for small volumes Consistent absorbency and wicking rates: ensures test-totest reproducibility Product manufactured in controlled environments from high-quality materials: minimizes the risk of false results due to sample contamination Low protein binding: high test sensitivity with minimal loss of analyte Naturally hydrophilic: rapid rewetting after prolonged storage Compatible: works with most housing styles Minimal leakage along the strip: minimizes the risk of contaminating test results Fast flowing: faster flow than cotton with lower sample retention Fusion 5 Fusion 5 is a proprietary, single-layer matrix membrane that can be used to replace traditional modular components from a lateral-flow testing kit. Features and benefits Fast flowing: quick and accurate results with wicking rates under 40 s for a 4 cm strip No blocking required: uses hydrophilic material High sensitivity: low background minimizes false-positive results Eliminates component contact issues: ensures accurate test results No assembly required: manufacturing is easy, fast, and efficient Fast test development: save time and money Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 164 Grade Thickness (μm @ 53 kPA) Wicking rate (s/4 cm) Water absorption (mg/cm2) Percent release of gold conjugate (after 90 s) Product code Rapid 24 348 20 52 80 8131-2250 Rapid 27 370 28 49 80 8132-2250 Fusion 5 370 38 42.3 > 94 8151-9915 Accuflow G 370 N/A 40 89 10537200 Standard 14 355 23.1 50.9 75 8133-2250 Standard 17 370 34.5 44.9 75 8134-2250 Custom roll, sheet, and disc sizes available upon request. Get in touch Unsure of which filtration material to use? Our team of experts are trained in troubleshooting filtration and can advise on which materials are most suitable for your assay. They can help identify the cause of and solution to any issues you might be having in diagnostic assay development. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Rapid 24 and Rapid 27 Treated bound glass fiber pad for good rewetting of membranes. Features and benefits Providing high absorption capacity and optimized as conjugate release material Rapid 24 — higher absorption capacity than Rapid 27 Rapid 27 — higher tensile strength than Rapid 24 Accuflow G Medium weight, treated bound glass fiber pad. Features and benefits High volume and medium absorption capacity that allows it to be used as a sample wick pad as well as for conjugate release. 165 Product name Size Thickness Wicking rate (s/4 cm) Product code Vivid 70 LFNC 25 mm × 100 m 190 to 230 μm includes 95 to 105 μm polyester support 64 to 90 VIV7025100R Vivid 70 LFNC 25 mm × 50 m 190 to 230 μm includes 95 to 105 μm polyester support 64 to 90 VIV702550R Vivid 70 LFNC 25 mm × 3 m 190 to 230 μm includes 95 to 105 μm polyester support 64 to 90 VIV702503R Vivid 90 LFNC 25 mm × 3 m 190 to 230 μm includes 95 to 105 μm polyester support 70 to 110 VIV902503R Vivid 90 LFNC 25 mm × 50 mc 190 to 230 μm includes 95 to 105 μm polyester support 70 to 110 VIV902550R Vivid 120 LFNC 25 mm × 3 m 190 to 230 μm includes 95 to 105 μm polyester support 95 to 135 VIV1202503R Vivid 120 LFNC 25 mm × 50 m 190 to 230 μm includes 95 to 105 μm polyester support 95 to 135 VIV1202550R Vivid 140 LFNC 25 mm × 100 m 190 to 230 μm includes 95 to 105 μm polyester support 120 to 160 VIV14025100R Vivid 140 LFNC 25 mm × 50 m 190 to 230 μm includes 95 to 105 μm polyester support 120 to 160 VIV1402550R Vivid 140 LFNC 25 mm × 3 m 190 to 230 μm includes 95 to 105 μm polyester support 120 to 160 VIV1402503R Vivid 180 LFNC 200 mm × 100 m 190 to 230 μm includes 95 to 105 μm polyester support 160 to 200 VIV180200100R Vivid 180 LFNC 205 mm × 100 m 190 to 230 μm includes 95 to 105 μm polyester support 160 to 200 VIV180205100R Vivid 180 LFNC 25 mm × 3 m 190 to 230 μm includes 95 to 105 μm polyester support 160 to 200 VIV1802503R Vivid 180 LFNC 25 mm × 100 m 190 to 230 μm includes 95 to 105 μm polyester support 160 to 200 VIV18025100R Vivid 180 LFNC 25 mm × 50 m 190 to 230 μm includes 95 to 105 μm polyester support 160 to 200 VIV1802550R Request a sample Request samples of membranes and paper for your immunoassay development. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Vivid lateral-flow nitrocellulose membranes With quality assured manufacturing and defined lot-to-lot consistency, our Vivid lateral-flow nitrocellulose (LFNC) membranes offer a reliable media for development, manufacturing, and implementation of lateral-flow diagnostic point of care tests. Features and benefits Vivid LFNC membranes are manufactured to ensure that these key parameters are tightly controlled with coefficients of variation (CV) between 5 and 10% minimizing detrimental effects on assay sensitivity, reproducibility, and reagent consumption. Performance Achieve high plasma yields with Vivid PS membrane The percent of plasma recovered from different volumes of blood does not depend on the blood volume applied to the media. Figure 1 shows that the separation time increases dramatically with increasing blood volume applied to the media. Figure 2 shows how Vivid PS membrane does not bind to clinically relevant protein biomarkers from plasma samples. All plasma samples were generated from the same sample of fresh EDTA blood spiked with Troponin I at 1 ng/mL. Protein concentration in each sample was measured in triplicate. 1.0 1.5 2.0 2.5 3.0 1.0 1.5 2.0 2.5 1.0 1.5 2.0 2.5 Low HTC Average HTC High HTC 100 80 60 40 20 0 Plasma recovery (% of available) Separation time (min) 20 15 10 5 0 Blood volume applied (in terms of void volumes) Fig 1. Plasma recovery (green columns) and separation time (orange rhombus) as a function of blood volume applied. ■ Recovered plasma ◆ Separation time 1.0 1.5 2.0 2.5 3.0 1.0 1.5 2.0 2.5 1.0 1.5 2.0 2.5 100 80 60 40 20 0 Plasma recovery (% of available) Separation time (min) 20 15 10 5 0 ■ Recovered plasma ◆ Separation time Fig 2. Troponin I concentration after filtration through Vivid PS membrane. Centr. control GR (1) GR (2) GF 1.6 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0 Troponin I concentration, ng/mL measured by ELISA 166 167 Item Description Capillary Flow Rate s/4 cm Thickness Product code FF80HP 20 mm × 50 m 60 to 100 200 µm 10547002 FF80HP 25 mm x 50 m 60 to 100 200 µm 10547003 FF80HP 210 mm × 297 mm 60 to 100 200 µm 13549206 FF80HP 60 (25) mm × 300 mm 60 to 100 200 µm 10547020 FF high performance (HP) membranes are part of the AE family, which are directly cast onto a plastic film. The FFHP membranes are a result of improved membrane casting procedures, which result in enhanced intra- and inter lot consistency and sharper lines. The surface is uniform without unincorporated nitrocellulose powder, and the fine structure fiber distribution provides large internal surfaces for binding proteins. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Nitrocellulose membranes The role of the membrane is to immobilize the capture molecules (typically antibodies) at the test and the control lines in a way that their binding properties for their respective target molecule are preserved. The membrane also guides the flow of the sample and the detection conjugate to the reaction area. To fulfil its role, a membrane must have a high and consistent protein binding capacity as well as porosity and wettability to allow a capillary flow of aqueous samples. The protein binding and capillary flow characteristics depend on the physical and chemical properties of the nitrocellulose membrane. To ensure consistent assay performance, test designers and manufacturers rely on test materials that perform consistently lot-to-lot. Cytiva products are manufactured to tightly controlled specifications to minimize product variability. Whatman FF80HP membranes A backed and highly consistent membrane for use with high density samples (e.g., undiluted serum) in a lateral-flow assay. Features and benefits Exceptional intra- and inter-lot consistency as demonstrated by a coefficient of variation (CV) for capillary rise of < 10% Improved assay consistency and reproducible limit of detection Reduced assay optimization costs compared to a less consistent membrane 168 Item Description Capillary Flow Rate s/4 cm Thickness Product code FF120HP 20 mm × 50 m 90 to 150 200 µm 10547006 FF120HP 25 mm x 50 m 90 to 150 200 µm 10547001 FF120HP 210 mm × 297 mm 90 to 150 200 µm 13549205 FF120HP 60 (25) mm × 300 mm 90 to 150 200 µm 10547021 FF120HP Plus 20 mm × 50 m 90 to 150 200 µm 10547126 FF120HP Plus 25 mm x 50 m 90 to 150 200 µm 10547125 FF120HP Plus 210 mm × 297 mm 90 to 150 200 µm 10547117 FF120HP Plus 60 (25) mm × 300 mm 90 to 150 200 µm 10547129 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Whatman FF120HP membranes A backed and highly consistent membrane for use with most sample types in a lateral-flow assay. Features and benefits Exceptional intra- and inter-lot consistency as demonstrated by a coefficient of variation (CV) for capillary rise of < 10% Improved assay consistency and reproducible limit of detection Reduced assay optimization costs compared to a less consistent membrane FF high performance (HP) membranes are part of the AE family, which are directly cast onto a plastic film. The FFHP membranes are a result of improved membrane casting procedures, which result in enhanced intra- and inter lot consistency and sharper lines. The surface is uniform without unincorporated nitrocellulose powder, and the fine structure fiber distribution provides large internal surfaces for binding proteins. 169 Item Description Capillary Flow Rate s/4 cm Thickness Product code FF170HP 20 mm × 50 m 140 to 200 200 µm 10547004 FF170HP 25 mm x 50 m 140 to 200 200 µm 10547005 FF170HP 210 mm × 297 mm 140 to 200 200 µm 13549204 FF170HP 60 (25) mm × 300 mm 140 to 200 200 µm 10547023 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Whatman FF170HP membranes A backed and highly consistent membrane for use with low viscosity samples in a lateral-flow assay. Features and benefits Exceptional intra- and inter-lot consistency as demonstrated by a coefficient of variation (CV) for capillary rise of < 10% Improved assay consistency and reproducible limit of detection Reduced assay optimization costs compared to a less consistent membrane The FFHP membranes are a result of improved membrane casting procedures, which result in enhanced intra- and inter lot consistency and sharper lines. The surface is uniform without unincorporated nitrocellulose powder, and the fine structure fiber distribution provides large internal surfaces for binding proteins. Lateral flow assay troubleshooting guide and how to switch diagnostic membranes Lateral flow assay troubleshooting and understanding molecular diagnostic membrane switch-out It’s a common scenario... your new diagnostic assay prototype is almost working the way you want it when an issue arises with a key component, the diagnostic membrane. You’re going to have to switch it out with an alternative, perhaps from another supplier, and that’s going to create substantial delays. Or is it? Switching to a different lateral flow assay membrane is never easy, but there are things you and your new supplier can do to make the switch as smooth as possible. Read on to find out more and discover key troubleshooting tips. Request a sample Interested in testing a nitrocellulose membrane for your lateral flow assay, request a free sample now! Sample pad/ blood separator Conjugate release Nitrocellulose membrane Absorption pad Test line Control line 170 171 Why might I need to switch-out a nitrocellulose membrane for a lateral flow assay? Lateral flow membranes form an integral part of diagnostic assays. Changing the nitrocellulose membrane in the later stages of assay development requires a complete revalidation of the test system, so developers will only switch membrane if there is a good reason. Ideally, they would also select a membrane and supplier that will help minimize any delays to market. Quality and supply chain issues are two of the most common reasons for changing a nitrocellulose membrane. You might find that the pore size on a production batch is inconsistent with that of the test batch, jeopardizing the performance of your entire lateral flow development assay. Or your supplier might run into manufacturing or logistical difficulties, prompting a membrane switch-out. How can a supplier simplify membrane switch-out for lateral flow assays? Switching to a different diagnostic assay membrane doesn’t need to be difficult, though it does require time and resources. Key decisions in any diagnostic membrane switch-out process include identifying a new supplier and selecting a material and grade that’s fit for purpose. Every supplier will be able to show you their ISO certifications, but can they help you get your diagnostic assay to market as quickly as possible? The aim is to find a supplier that can demonstrate a history of delivering high-flow nitrocellulose membranes while matching the specifications you give them, supporting you from the first day to the last. One thing to look out for, regardless of whether a nitrocellulose membrane switch-out is due to quality or supply issues, is whether this potential new supplier can provide accurate comparison tables and knowledgeable people to help in finding a suitable replacement assay membrane. They should know their products best, and be able to help you streamline the switch-out process. If quality improvements are the primary driver, the million-dollar question is whether you have successfully identified the material property needing improvement. For example, when liquid flow is inconsistent or slow, does the solution lie in the material’s chemistry (e.g. hydrophilicity) or in its physical properties (e.g. density, pore size, etc.). The supplier’s in-house experts can work with you to confirm the existing issue, find any potential challenges, and provide appropriate test samples. Working closely with an experienced supplier can relieve some of the burden on the lateral flow assay developer, simplifying the nitrocellulose membrane switch-out process. What can I do to ease the process of nitrocellulose membrane switching? Membrane material specifications: Raw material sources and proprietary additives for glass fiber and paper pads vary between suppliers, leading to differences in performance that might need some adjustments to equalize. Assay antibody chemistry: Although the risk is small, antibody chemistry might need adjusting to make sure the later flow assay’s sensitivity and reliability is unaffected by differences in nitrocellulose membrane surfactant. Other assay redevelopment: Exchanging the lateral flow test membrane will most likely require a redevelopment of the dispensing blocking buffer systems and the conjugate pad chemistry. While you might think addressing these points could add further delays, each is manageable with a well-informed supplier, and well worth considering from the start. Download Download our ‘Considerations for lateral flow membrane selection’ infographic. Download Download our 10 top tips for lateral flow assay development infographic. 172 What are some other lateral flow assay troubleshooting solutions? Should you still run into issues during membrane switch-out or revalidation, here are a few common challenges and (potential) quick fixes our in-house experts have encountered and solved. Hopefully you find them useful. Request a sample Looking to try a new membrane in your existing solution? Building a new lateral flow device? Get a free sample of our Fast Flow High Performance nitrocellulose backed membrane. Challenge Possible solution >Uneven lines or dots Use membranes with different pore size Reduce dispensing volume of reagent Increase protein concentration of reagent Check dispensing buffer composition Check dispensing process False positive signals Modify buffer in conjugate pad/solution (e.g. pH, salt concentration, or surfactant concentration) Change conjugated protein False negative signals Modify buffer in conjugate pad/solution (e.g. pH, salt concentration, or surfactant concentration) Change conjugated protein Use membrane with smaller pore size Increase sample volume Uneven liquid fronts of migrating sample Check membrane shelf life Use membrane with different/more surfactant Check whether the relative humidity is very low Increase surfactant concentration in conjugate pad 173 Item Description Capillary Flow Rate s/4 cm Thickness Product code FF80HP Plus Thick 20 mm × 50 m 60 to 100 235 µm 10547155 FF80HP Plus Thick 25 mm x 50 m 60 to 100 235 µm 10547156 FF80HP Plus Thick 210 mm × 297 mm 60 to 100 235 µm 13547204 FF80HP Plus Thick 60 (25) mm × 300 mm 60 to 100 235 µm 10547154 FF120HP Plus Thick 20 mm × 50 m 90 to 150 235 µm 10547152 FF120HP Plus Thick 25 mm × 50 m 90 to 150 235 µm 10547149 FF120HP Plus Thick 210 mm × 297 mm 90 to 150 235 µm 13547200 FF120HP Plus Thick 60 (25) mm × 300 mm 90 to 150 235 µm 13547202 FF170HP Plus Thick 20 mm × 50 m 130 to 210 235 µm 10547153 FF170HP Plus Thick 25 mm x 50 m 130 to 210 235 µm 10547147 FF170HP Plus Thick 210 mm × 297 mm 130 to 210 235 µm 13547201 FF170HP Plus Thick 60 (25) mm × 300 mm 130 to 210 235 µm 13547203 Download Download our infographic: Considerations for membrane selection. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Whatman FFHP plus thick membranes A thicker nitrocellulose membrane with added surfactant, creating a large-pore, backed membrane (235 µm thick) for easy replacement of other membranes in lateral-flow assays with minimal optimization work needed. Features and benefits Reliable intra- and inter-lot consistency as demonstrated by a coefficient of variation (CV) for capillary rise of < 10% Enhanced consistency in application with minimal amount of unincorporated nitrocellulose powder (often referred to as “sticky dust”) on the surface of the membrane Reduced assay optimization costs, in terms of cost and time, as compared to a less consistent membrane Easily swapped into existing lateral-flow assays without the need for a large redevelopment FFHP plus thick membranes are part of the Whatman FFHP family of nitrocellulose membranes, which are directly cast onto a plastic film. The membranes are a result of an improved casting procedure, which results in enhanced intra- and inter-lot consistency and sharper lines. The surface is uniform with minimal amounts of unincorporated nitrocellulose powder, while the fine structure fiber distribution provides large internal surfaces for binding proteins. 174 Fig 2. Thickness and flow rates for (A) FF80HP Plus thick membranes, (B) FF120HP Plus thick membranes, and (C) FF170HP Plus thick membranes. Surface quality AdttdbthiiFi3thffthbiifithiiltfThickness (μm) Flow rate (4 cm/s) FF80HP Plus thick Thickness (μm): 215–255 Flow rate (4 cm/s): 60–100 FF120HP Plus thick Thickness (μm): 215–255 Flow rate (4 cm/s): 90–150 FF170HP Plus thick Thickness (μm): 215–255 Flow rate (4 cm/s): 130–210 (A) (B) (C) Mean LSL USL 3sp 3sp Target 0 5 10 15 210 215 220 225 230 235 240 245 250 255 Mean LSL USL 3sp 3sp Target 0 10 20 30 40 50 60 65 70 75 80 85 90 95 100 Mean LSL USL 3sp 3sp Target 0 5 10 15 215 220 225 230 235 240 245 250 255 260 Mean LSL USL 3sp 3sp Target 0 5 10 15 90 100 110 120 130 140 150 Mean LSL USL 3sp 3sp Target 0 2 4 6 8 10 210 215 220 225 230 235 240 245 250 255 Mean LSL USL 3sp 3sp Target 0 5 10 15 20 120 140 160 180 200 220 240 Based on the quality testing of a group of validated samples in terms of thickness and flow rate, our data demonstrates that our membranes are manufactured within the promised range and only deviate approximately ± 10% in terms of thickness and flow rate from the given norm. Fig 1. Thickness and flow rates for (A) FF80HP Plus thick membranes, (B) FF120HP Plus thick membranes, and (C) FF170HP Plus thick membranes. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 175 Item Description Capillary Flow Rate s/4 cm Thickness Product code AE99 25 mm × 50 m 120 to 160 120 μm 10548081 AE99 20 mm × 100 m 120 to 160 120 μm 10539706 AE99 25 mm × 50 m 120 to 160 120 μm 10549916 Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Whatman AE membranes A range of unbacked nitrocellulose membranes for use in a lateral-flow assay. Features and benefits AE membranes are unbacked, allowing either belt or air side of the membrane to be used Enables assay suitability tests of both air and belt side of the membrane Constructed of 100% nitrocellulose Constructed of 100% nitrocellulose, the AE family of membranes offers a higher level of purity and performance compared to posttreated materials. The AE membranes have been used extensively since the development of the original lateral-flow tests and have become a standard for manufacturers worldwide. There is a long history of success and experience for the optimization of these products in assays. AE membranes are unsupported, which means either the belt or air side of the membrane can be used. 176 Our services range from component customization to complete development of your molecular or immunoassay. We have you covered from idea to commercialization. “ 177 Item Thickness (mm) Thickness (μm) Blood volume (μL/cm2) Membrane void volume Plasma separation time (minutes) Plasma recovery (%) Product code Vivid GF PSM 12.99 330 20 1× ≤ 2 ≤ 60 T9EXPPA0200S00A Vivid GX PSM 12.99 330 20 to 30 1 to 1.5× ≤ 2 ≤ 60 T9EXPPA0200S00X Vivid GR PSM 12.99 330 40 to 50 2 to 2.5× ≤ 2 ≤ 80 T9EXPPA0200S00R Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Vivid plasma separation membrane The Vivid plasma separation membrane (PSM) utilizes a patented process where a highly asymmetric membrane is specifically engineered for one-step plasma separation from whole blood without the use of centrifugation. Cellular components of the blood (red cells, white cells, and platelets) can be captured in the larger pores without lysis, while the plasma flows down into the smaller pores on the downstream side of the membrane. Features and benefits High plasma yield: Achieve high quality plasma similar in HPLC and SDS-PAGE profiles to traditional centrifuged plasma in less than two minutes with ≥ 80% plasma yield, lowering the amount of starting whole blood needed Low analyte binding: Low, nonspecific binding of common diagnostic biomarkers and target analytes Dependable performance: Efficient removal of the cellular components of whole blood, including red cells, white cells, and platelets Low lysis: Hemolysis levels significantly lower than glass fiber media generated plasma Device integration: Compatible with point-of-care (POC) and point-of-use (POU) diagnostic platforms such as lateral-flow test strips and microfluidics 178 Nylon membranes Biodyne™ and Immunodyne™ nylon transfer membranes Nylon membranes have been recognized as the gold standard for nucleic acid detection for over 35 years. High sensitivity, low background, and lot-to-lot consistency for all radioactive and nonradioactive detection methods is provided. The intrinsically hydrophilic nature of the membrane results in easy wetting across the membrane. When subjected to multiple cycles of hybridization, stripping, and reprobing, the membranes remain stable and durable as they will not crack, shrink, or tear. We have five chemistries available that provide versatile adsorption properties: Amphoteric nylon 6,6: The net charge of this membrane can be modulated by changes in pH. The binding mechanism for this membrane is through hydrophobic and electrostatic interactions. Features and benefits Ideal for single probe or multiple rehybridizations and in applications where background is troublesome. Nucleic acids can be immobilized to this membrane via UV crosslinking and baking. Well suited for nucleic acid dot blots, gene probe assays, and DNA fingerprinting. Positively-charged nylon 6,6: The pore surfaces of this membrane are populated by a high density of quaternary ammonium groups. It offers high sensitivity in nucleic acid detection applications. The binding mechanism for this membrane is through electrostatic interactions. Features and benefits Nucleic acids can be immobilized to this membrane via UV crosslinking and baking although it is not required. Well suited for nucleic acid dot blots, DNA fingerprinting, and colony/plaque lifts. Positively-charged Nylon 6,6 with high isoelectric point: This membrane has an extremely high isoelectric point that provides greater sensitivity than the amphoteric nylon while exhibiting lower background than the standard positively-charged nylon membrane in certain nonradioactive detection systems. The binding mechanism for this membrane is through electrostatic interactions. Features and benefits Our most sensitive nylon membrane for nucleic acid detection. Nucleic acids can be immobilized to this membrane via UV crosslinking and baking although it is not required. Well suited for nucleic acid dot blots and DNA fingerprinting. Negatively-charged nylon 6,6: The surface of this membrane is covered with a high density of carboxyl groups that can be derivatized for protein attachment. The binding mechanism for this membrane is through electrostatic interactions. Features and benefits Well suited for protein immobilization, ELISAs, and affinity purification via ligand attachment. Modified nylon 6,6: This membrane is surface modified through proprietary chemistry that allows for covalent binding and stable ligand immobilization. The membrane is preactivated to form covalent linkages with nucleophilic groups found on proteins and other biological macromolecules. Primary reactivity is with amine groups at neutral pH. Features and benefits The microporous structure of the membrane provides an available immobilization area of up to 300 cm2 for each cm2 of planar membrane. Intrinsically hydrophilic and exhibits instantaneous wetting on contact with low ionic strength aqueous solutions. High capillarity and rapid absorbent wicking are obtainable without the necessity of surface modifying wetting agents. Prewetting is not required prior to contact with ligand solutions. This membrane offers greater binding capacity for proteins than traditional nonporous solid phase surfaces. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 179 Grade Base material Pore size Thickness Product code Biodyne A membrane Amphoteric supported nylon 6,6 0.2 μm 139.7 to 177.8 μm BNRG810S Biodyne A membrane Amphoteric supported nylon 6,6 0.45 μm 144.8 to 170.2 μm BNXG810S Biodyne A membrane Amphoteric supported nylon 6,6 1.2 μm 139.7 to 177.8 μm BNNF810S Biodyne A membrane Amphoteric supported nylon 6,6 0.2 μm 60113 Biodyne A membrane Amphoteric supported nylon 6,6 0.45 μm 60106 Biodyne A membrane Amphoteric supported nylon 6,6 0.45 μm BNXF810S Biodyne A membrane Amphoteric supported nylon 6,6 0.45 μm BSP0533 Biodyne A membrane Amphoteric supported nylon 6,6 0.2 μm BNRGR500 Biodyne A membrane Amphoteric supported nylon 6,6 0.2 μm BNRGR15240 Biodyne B membrane Positively charged supported nylon 6,6 0.45 μm 144.8 to 170.2 μm BNBZF810S Biodyne B membrane Positively charged supported nylon 6,6 0.8 μm 139.7 to 177.8 μm BNHZF810S Biodyne B membrane Positively charged supported nylon 6,6 0.45 μm 60200 Biodyne B membrane Positively charged supported nylon 6,6 0.45 μm 60201 Biodyne B membrane Positively charged supported nylon 6,6 0.45 μm 160 to 200 60208 Biodyne B membrane Positively charged supported nylon 6,6 0.45 μm 160 to 200 60207 Table continued > Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 180 Grade Base material Pore size Thickness Product code Biodyne B membrane Positively charged supported nylon 6,6 0.45 μm 139.7 to 177.8 μm BNBZH50001 Biodyne B membrane Positively charged supported nylon 6,6 0.45 μm 144.8 to 170.2 μm BSP0616 Biodyne B membrane Positively charged supported nylon 6,6 0.45 μm 139.7 to 177.8 μm BNBZH50006 Biodyne B membrane Positively charged supported nylon 6,6 0.45 μm BSP0687 Biodyne B membrane Positively charged supported nylon 6,6 0.45 μm BNBZH3R Biodyne C membrane Negatively charged supported nylon 0.45 μm 279.4 to 330.2 μm (dual layer measurement) BNBCH810S Biodyne C membrane Negatively charged supported nylon 1.2 μm 139.7 to 177.8 μm BNNCH810S Biodyne C membrane Negatively charged supported nylon 0.45 μm 60314 Biodyne C membrane Negatively charged supported nylon N/A 60336 Biodyne C membrane Negatively charged supported nylon 0.45 μm 60315 Biodyne C membrane Negatively charged supported nylon 1.2 μm BSP0193 Biodyne Plus membrane Positively charged supported nylon 6,6 with high isoelectric point 0.45 μm 144.8 to 170.2 μm BNHZF810S Biodyne Plus membrane Positively charged supported nylon 6,6 with high isoelectric point 0.45 μm 60400 Biodyne Plus membrane Positively charged supported nylon 6,6 with high isoelectric point 0.45 μm 60406 Biodyne Plus membrane Positively charged supported nylon 6,6 with high isoelectric point 0.45 μm 60404 Immunodyne ABC membrane Modified nylon 6,6 0.45 μm 279.4 to 330.2 μm (dual layer measurement) BC045H810S Immunodyne ABC membrane Modified nylon 6,6 1.2 μm 139.7 to 177.8 μm BC120H810S Note: Exact binding capacities are difficult to determine because biomolecules tend to layer on the membrane surface when applied at very high concentrations. Highest effective loads for nucleic acids and proteins are typically less than 100 μg/cm2 . Higher loads usually do not result in higher activity due to steric hindrance or other phenomena. For molecular detection, maximum signal is usually detected with less than 10 μg/cm2 , or 1 μg/μL, of applied solution. Biomolecules tend to concentrate near the surface of nylon membranes as they bind quickly to the nylon membranes and separate (similar to chromatography) from the solvent carrier. Because of this, binding to the membrane depends less on the total internal surface area of the membrane than expected. Spotting biological levels of nucleic acid will produce similar binding to 0.2 or 1.2 μm nylon membranes, despite the much greater internal surface area of the smaller pore size membrane. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 181 Product Size Pore Application Product code Amersham Hybond P 0.2 1 roll, 26 cm × 4 m 0.2 μm Standard colorimetric and chemiluminescence detection methods for proteins of < Mr 20 000. 10600021 Amersham Hybond P 0.45 1 roll, 30 cm × 4 m 0.45 μm Chemiluminescence and fluorescence detection methods for proteins of > Mr 20 000. 10600023 Amersham Hybond LFP 1 roll 25.4 cm × 4 m 0.2 μm Fluorescence Western blotting applications. 10600022 Amersham Hybond P 0.2 10 units, 8 × 9 cm 0.2 μm PVDF membranes preassembled with 2 × 3 mm Chr blotting papers 10600122 Amersham Hybond P 0.45 10 units, 8 × 9 cm 0.45 μm PVDF membranes preassembled with 2 × 3 mm Chr blotting papers 10600121 Amersham Hybond LFP 0.2 10 units, 8 × 9 cm 0.2 μm PVDF membranes preassembled with 2 × 3 mm Chr blotting papers 10600123 Read more Read more about choosing the right nylon membrane. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection PVDF blotting membranes Amersham Hybond™ membranes The Western blot (or immunoblot) technique has been a fundamental in protein analysis since the 1970s when it was first discovered that biomolecules could be spotted directly onto membranes (spot ELISA or DNA dot blots), or transferred from gels (Southern blots, Northern blots, Western blots). Despite the now routine application of Western blotting, successful results can depend on several factors including optimal membrane selection for the desired detection method and protocol. Features and benefits High tensile strength and low background signal Low membrane burn-through Compatibility with a wide selection of detection methods without compromising quality. 182 Product Pore size Thickness Leukocyte immobilization Leukocyte retention post wash Hold-up Product code Leukosorb B Media 8” × 10” sheets 8 µm 355.6 to 558.8 µm 40 to 80% 70 to 80% 40 to 70 µL/cm2 BSP0669 Note: Custom roll, sheet, and disc sizes available upon request. Note: Unsure of which filtration material to use? Our team of experts are trained in troubleshooting filtration and can advise on which materials are most suitable for your assay. They can help identify the cause of and solution to any issues you might be having in diagnostic assay development. Red cells, white cells, platelets and plasma Red cells, platelets and plasma pass through White cells are retained in filter media Get in touch Contact your local Cytiva representative to discuss your needs. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Leukosorb™ media Leukosorb™ leukocyte removal medium is a highly wettable, fibrous matrix designed for use in procedures requiring isolation of leukocytes from whole blood samples, which can then be prepared for use in polymerase chain reaction (PCR) applications. Features and benefits Specifically designed for the isolation of leukocytes from whole blood Choice of system — whole blood samples can be applied to the top of membrane and allowed to pass through the filter matrix either by gravity or pressure. If using a pressurized filtration system, care must be employed not to lyse the red blood cells resulting in hemoglobin contamination. The leukocytes are isolated from the sample and become trapped within the fibrous matrix of the membrane. 183 Item Thickness (mm) Thickness (μm) Blood volume (μL/cm2) Membrane void volume Plasma separation time (minutes) Plasma recovery (%) Product code Vivid GF PSM 12.99 330 20 1× ≤ 2 ≤ 60 T9EXPPA0200S00A Vivid GX PSM 12.99 330 20 to 30 1 to 1.5× ≤ 2 ≤ 60 T9EXPPA0200S00X Vivid GR PSM 12.99 330 40 to 50 2 to 2.5× ≤ 2 ≤ 80 T9EXPPA0200S00R Note: Custom roll, sheet, and disc sizes available upon request. Note: Unsure of which filtration material to use? Our team of experts are trained in troubleshooting filtration and can advise on which materials are most suitable for your assay. They can help identify the cause of and solution to any issues you might be having in diagnostic assay development. Get in touch Contact your local Cytiva representative to discuss your needs. Download Download our infographic: 10 tips for lateral-flow development. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection Vivid plasma separation membranes The Vivid plasma separation membrane (PSM) utilizes a patented process where a highly asymmetric membrane is specifically engineered for one-step plasma separation from whole blood without the use of centrifugation. Cellular components of the blood (red cells, white cells, and platelets) can be captured in the larger pores without lysis, while the plasma flows down into the smaller pores on the downstream side of the membrane. Features and benefits High plasma yield: Achieve high quality plasma similar in HPLC and SDS-PAGE profiles to traditional centrifuged plasma in less than two minutes with ≥ 80% plasma yield, lowering the amount of starting whole blood needed Low analyte binding: Low, nonspecific binding of common diagnostic biomarkers and target analytes Dependable performance: Efficient removal of the cellular components of whole blood, including red cells, white cells, and platelets Low lysis: Hemolysis levels significantly lower than glass fiber media generated plasma Device integration: Compatible with point-of-care (POC) and point-of-use (POU) diagnostic platforms such as lateral-flow test strips and microfluidics 184 Detection So, with the rapid evolution in immunodiagnostics, what does the next generation of assays look like? One key technology enabling the next generation are microfluidics. Microfluidics is used in a variety of application areas, including antibody characterization. These systems use immobilized antibodies on surface plasmon resonance (SPR) chips to study analyte interaction but can also be used the other way around to look for biomarkers in samples in real time. The OPKO Health PSA screening assay mentioned earlier uses this approach for biomarker identification, and there are many microfluidics-based devices already on the market. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 185 Training and workshops As part of Cytiva Diagnostic Services, we provide hands-on training and education, on-site at our Dassel, Germany facility or off-site at locations convenient to you. Our training and education services maximize our collaboration and help you take your assay development forward and be fully informed for your next project. On-site access and participation during all development activities Test development lab for proof-of-concept Support staff at your site for start-up activities Access to process, equipment, and development training Site safety training Classroom, on-site, and online education Workshop format to work through specific issues Reagent characterization Commercially available antibodies are typically poorly characterized, and supplier statements on intended use are not necessarily reliable. Surface plasmon resonance technology (SPR) can be a valuable tool for the quick down-selection of specific binding reagents for lateral-flow assays. Contract antibody manufacturers may have to be asked to provide all supernatants of all hybridoma cell lines they created to their customer for SPR analysis. SPR also provides an option for quality control of incoming goods. Concentration and kinetic parameters can be determined within a short period of time. Full test and conjugate development We offer laboratory infrastructure, technical expertise, training, and consultation services to diagnostic developers looking to convert an idea into a rapid diagnostic test or access additional capacity to scale up or manufacture a diagnostic assay successfully. Our full test development services include: Method development Method transfer Method qualification Troubleshooting Full outsourcing of development Learn more Learn more about our lateral flow immunoassay development services. Learn more Learn more about our Biacore SPR — surface plasmon resonance interaction analysis. Process Cytiva products Cytiva services Biomarker identification Protein selection Reagent purification Membrane family selection Sample collection Chromatography Detection 186 Diagnostic services 187 Building on over 40 years’ experience in the life sciences industry, we integrate with the teams of biopharma companies developing diagnostic kits and assays by sharing our knowledge, experiences, and project updates as part of a rigorous process to move projects to commercialization with minimal risk. Value added services We put our diagnostic team to work on your solution to address workflow needs and changes. Lyophilization to remove cold-chain requirements Conjugation and chemistry services to remove steps from your workflow and enhance your product Kitting and contract manufacturing support Diagnostic services: your assay is in safe hands The benefits of lyophilization in assay kit development Freeze drying, lyophilization, and cryodesiccation are all terms used to describe the process of removing water from a sample at low temperature. This low-temperature dehydration process maximizes product or sample stability and shelf life, maintains chemical or biological function, and enables easier transportation and storage compared to a cold chain. The process has applications in assay kit development, where it enables room-temperature shipping and storage of reagents and complete assays. Sample stabilization by lyophilization enables the production of predispensed, single-dose reagents, which help: simplify assay setup increase assay robustness and reliability reduce the risk of sample contamination Diagnostic services 188 What is lyophilization? Lyophilization is a freeze-drying process (also known as cryodesiccation) that removes water from a sample or product by freezing and placing it under a vacuum. It relies on a phenomenon known as the triple point of water — the combination of temperature and pressure where three phases of water (solid, liquid and gas) can co-exist and are equally stable. The lyophilization process typically involves three stages (Fig 1): freezing primary drying secondary drying A product or sample is first frozen with cryoprotective excipients, either in a dedicated freeze dryer or in a freezer at temperatures of approximately -40°C (below the triple point of water). Excipients provide a layer of protection during the freeze-drying process and help maintain stability of the intended function over time. The optimum combination and concentration of excipients are unique to each product or sample. Gradual freezing would usually provide maximum efficiency for lyophilization, but this process forms large ice crystals. The large ice crystals can disrupt and damage materials that are structurally sensitive, such as cells. In these cases, the freezing step employs a rapid cooling profile to minimize ice crystal size. The primary drying phase involves reducing the pressure of the surrounding environment below that of the triple point of water. The application of a small amount of heat then energizes the free ice crystals to undergo sublimation, transitioning directly from a solid to a gas phase. The primary drying step removes approximately 95% of the water and results in the product or sample transforming from a glassy frozen state to a largely dry powder. The final step removes any remaining unfrozen water molecules adsorbed to the product or sample. The temperature is increased slightly, disrupting interactions between water molecules and the frozen material, removing them through desorption. The result is a lyophilized powder that is much lighter in mass and smaller in volume than the original product or sample. Readying this lyophilized powder for use then simply involves adding sterile, distilled water or an appropriate buffer to form a solution in a process called reconstitution. The challenges in long-term sample storage Often, samples that require long-term storage in the biotechnology and pharmaceutical industries are highly labile. Application of heat for dehydration can, therefore, have consequences in terms of sample stability and quality. Enzymes, for example, are used in many different assay kits and are often dehydrated to extend shelf life and ease transportation. These proteins have precise macromolecular structures required for their specific biological function. Heat stress would disrupt the noncovalent bonds and interactions that maintain these structures and in turn disrupt the in vivo function. Not dehydrating samples intended for long-term storage is also not a viable option in many cases. Water can act as a powerful solvent and has a dramatic change in chemical structure when frozen. As a result, water can break covalent bonds found in biological macromolecules and, because the solid structure differs vastly from the liquid, those molecules can precipitate. Fig 1. The process of lyophilization involves three stages that remove water from a sample by freezing it and placing it under specific vacuum pressures and temperatures. 20 10 0 -10 -20 -30 -40 -50 -60 Shelf temperature (°c) 95% to 99% water Vacuum 5% to 10% water 1% to 2% water Diagnostic services 189 How lyophilization addresses sample storage challenges Although it is a complicated process, lyophilization has become standard practice in many industries, including pharmaceutics, biotechnology, and agriculture. Given the adverse effects of heat and water, dehydration by lyophilization offers several advantages, from improved sample stability and purity to increased shelf life and reduced costs. By removing the need to dehydrate by heating, lyophilization provides a convenient and safe method for long-term storage of lab samples and pharmaceutical products while preserving their activity. As well as increasing shelf life, freeze-drying samples also reduces their weight and volume, helping cut down on shipping costs and environmental impact. Eliminating the need for shipping procedures designed to maintain sample stability, such as dry ice, also simplifies logistics and helps further reduce costs. These benefits are especially advantageous when transporting samples and assays to developing countries with limited facilities and budgets. Long transportation distances and often hot and humid environments would otherwise put a substantial strain on cold chain transport and storage. With lyophilization, a temperature-controlled chain is not necessary. An example of transport without a cold chain happened during World War II, which saw one of the first use cases of freeze-drying as a commercial technique (5). Lyophilization was used to render blood plasma and penicillin chemically stable and viable without refrigeration. As a result, both could be transported to their intended recipients regardless of cold chain transport availability. This process is still in use today. A complete, stabilized assay in a single tube In addition to the practical and logistical benefits to shipping, costs, and shelf life, lyophilization offers scientists reliable, efficient, and accurate research tools in the form of individual reagents and complete, ready-to-use formulations in assay kits. Through lyophilization, the individual components of an assay kit can be combined in a single tube. The result is a predispensed, single-dose reagent that vastly simplifies assay setup (Fig 2). In many cases, and particularly for pharmaceutical and biological products, excipients will be critical in formulations to help maintain the stability of the macromolecular structure (6, 7). Enzymes, for example, are still susceptible to destabilization during dehydration, losing activity even without the addition of heat. Excipients, such as amino acids and nonaqueous solvents, can act as cryoprotectants, minimizing this dehydration stress. Using this lyophilization approach, researchers do not need to individually pipette microliter quantities of potentially expensive assay reagents, greatly simplifying their workflows. Fewer pipetting steps and less sample handling also help minimize training requirements, reduce costs, and save time. The robustness and reliability of an assay are also improved along with data quality, and the risk of sample contamination is minimized. Fig 2. A complete assay in a single tube. Example of components for an amplification-based custom Lyo-Stable services formulation. Other user-defined components Enzyme(s) Primers/ probes Ready-to-gostabilization Propietory excipient mix Buffer dNTPs MgCI2 Read more Read the article in full. Diagnostic services 190 Custom solutions Using our core technology of proven diagnostic components, we can work with you to create bespoke solutions to fit your needs. Product customization: from pack sizes to bespoke product changes Product development: custom versions of our high-quality components Custom multiwell plate design Download Download our infographic: 10 essentials when choosing a collaborator. Start your project today Diagnostic services Sharing knowledge and experience Whether you are looking to complete one aspect of contract development or do a full test development, our experienced team is happy to work on your project. Classroom, on-site, and online education: process, equipment, or application focused Workshops and consultation services: including SPR analysis in diagnostic test development and bespoke projects Contract development services 191 References 1. Sharma R, Gale BK. Electrochemical quantification of DNA using aluminum oxide membranes. Procedia Engineering. 2011;25:713-716. Doi:10.1016/j.proeng.2011.12.176 2. Mueller JA, Culley AI, Steward GF. Variables influencing extraction of nucleic acids from microbial plankton (viruses, bacteria, and protists) collected on nanoporous aluminum oxide filters. Applied and Environmental Microbiology. 2014;80(13):3930-3942. Doi:10.1128/ aem.00245-14 3. Asano K, Ono A, Hashimoto S, Inoue T, Kanno J. Screening of endocrine disrupting chemicals using a surface plasmon resonance sensor. Analytical Sciences. 2004;20(4):611- 616. Doi:10.2116/analsci.20.611 4. Kuzuhara T, Sei Y, Yamaguchi K, Suganuma M, Fujiki H. DNA and RNA as new binding targets of Green Tea Catechins. Journal of Biological Chemistry. 2006;281(25):17446-17456. Doi:10.1074/jbc.m601196200 5. Sailliol A, Martinaud C, Cap AP, et al. The evolving role of lyophilized plasma in remote damage control resuscitation in the French Armed Forces Health Service. Transfusion. 2013;53(S1). doi:10.1111/trf.12038 6. Bansal A, Lale S, Goyal M. Development of lyophilization cycle and effect of excipients on the stability of catalase during lyophilization. International Journal of Pharmaceutical Investigation. 2011;1(4):214. doi:10.4103/2230-973x.93007 7. Wang W. Lyophilization and development of Solid Protein Pharmaceuticals. International Journal of Pharmaceutics. 2000;203(1-2):1-60. doi:10.1016/s0378-5173(00)00423-3 192 Cytiva and the Drop logo are trademarks of Life Sciences IP Holdings Corporation or an affiliate doing business as Cytiva. 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