Streamlining End-to-End Testing for Optimized Monoclonal Antibody Production

Since the approval of the first monoclonal (mAb) for clinical use in 1986, they have become a key therapeutic for a range of diseases including cancers and autoimmune disorders.1,2 All mAbs are subject to rigorous quality regulations to ensure safety and purity during clinical use. As such, products must be tested for contaminants at multiple stages during development and manufacture. Inefficient quality control (QC) protocols can result in costly delays or failure to meet regulatory guidelines. This infographic will explore how robust, sensitive kit-based QC solutions can be applied across the mAb production workflow to save time and resources. Successful production and approval of mAbs depend on robust characterization – the final product’s safety, efficacy, purity, quality and potency need to be proved. Arising from various sources and organisms, contaminants and impurities occur at all stages of the mAb development and manufacturing process. Therefore, testing must be carried out throughout the process, from research and development to lot release. Helping to ensure quality at every stage Environmental monitoring Microbial identification Product safety Mycoplasma & viral detection Product quality Impurity analysis Streamlining End-to-End Testing for Optimized Monoclonal Antibody Production Proving quality in mAb workflows Minimized in-house staffing requirements In-house testing for QC requires complex optimization and validation to ensure results are in line with regulatory guidelines. This can be time-consuming and result in high long-term costs, and it demands extensive expertise. In comparison, commercial kits and analysis software can offer a far more efficient solution: Advantages of commercial testing kits Less than half the time to produce a qualified method Method creation and maintenance provided Designed to meet regulatory guidelines, such as cGMP and ICH Q2 (R1) guidance for analytical procedures. Can help ensure data lineage and traceability for 21 CFR Part 11 compliance Access to support from a dedicated team of specialists, from implementation to validation Reduced overhead costs References 1. Todd PA, Brogden RN. Muromonab CD3. Drugs. 1989;37(6):871-899. doi:10.2165/00003495-198937060-00004 2. Manis JP. Overview of therapeutic monoclonal antibodies. UpToDate. Wolters Kluwer. https://www.uptodate.com/contents/overview-oftherapeutic-monoclonal-antibodies/print. Updated April 10, 2024. Accessed June 6, 2024 3. Barone PW, Wiebe ME, Leung JC, et al. Viral contamination in biologic manufacture and implications for emerging therapies. Nat Biotechnol. 2020;38:563-572. doi:10.1038/s41587-020-0507-2 4. US Food and Drug Administration. Sterile Drug Products Produced by Aseptic Processing — Current Good Manufacturing Practice: Guidance for Industry. https://www.fda.gov/regulatory-information/search-fda-guidance-documents/sterile-drug-products-produced-aseptic-processingcurrent-good-manufacturing-practice. Published October 2004. Updated 4 May, 2020. Accessed June 10, 2024. PCR assay kits may offer expert efficiency, but a complete workflow can take turnaround and reliability to the next level. Thermo Fisher Scientific offers end-to-end testing with a range of integrated solutions. From complementary sample preparation and DNA/RNA extraction, through the SEQ family of assays to technology platforms, including qPCR and dPCR instruments with integrated software for streamlined study set up and data analysis, this workflow enables rapid turnaround of accurate, actionable results. An integrated solution for a complete workflow Propel your innovations forward Prepare sample Set up reaction Run assay Analyze results Thermo Scientific™ Pharma KingFisher™ with PrepSEQ™ or Manual w/PrepSEQ™ kit DNA/RNA extraction < 2.5 hours Real-time PCR 1.5–2.5 hours Applied Biosystems™ resDNASEQ, MycoSEQ & ViralSEQ™ Kits Applied Biosystems™ QuantStudio™ 5 or 7500 Fast Real-Time PCR System Applied Biosystems™ AccuSEQ™ Software Bacterial identification Fungal identification Residual DNA quantitation Virus detection and quantitation Mycoplasma detection Bacterial and fungal identification Scale up to production Cell culture development Harvest and collection Bulk storage and final fill Purification Enviro nment al mon itorin g Produ ct saf ety Product quality Virus detection and quantitation Contamination by viruses, such as mouse minute virus and vesivirus, can have significant consequences for mAb safety. Although viruses can be difficult to detect in cell culture, virus-specific rapid PCR testing is the best option for identifying viral contaminants early in the mAb workflow, preventing impacts further downstream.3 The Applied Biosystems™ ViralSEQ™ detection and quantitation kits allow the detection of specific viruses that pose risks of contamination. • No cross-reactivity to non-target species • Same day actionable results, within 5 hours, enabling in-process testing • Highly sensitive detection using Applied Biosystems™ TaqMan™ chemistry for broad dynamic range • Includes discriminatory positive/ extraction DNA controls for a reduced risk of false positives Bacterial and fungal identification Identifying microorganisms to a species level is a minimal requirement to comply with good manufacturing practices. Genotypic identification is considered more accurate and precise than phenotypic methods, and so is the more reliable choice for mAb workflows.4 The Applied Biosystems™ MicroSEQ™ Rapid Microbial Identification System combines PCR and DNA sequencing technologies for highly accurate environmental monitoring. • Comprehensive, validated library of bacteria and fungi species—more than 12,000 strain types • Regulatory compliant with approaches recommended by global regulatory agencies • Rapid results in ~5 hours from colony to identification, allowing in-process testing • Worldwide technical support network, including training and implementation guidance Mycoplasma detection Mycoplasmas can interfere with cell health during mAb manufacture, and negatively affect product safety and quality. Early detection of mycoplasma can help prevent costly manufacturing delays or remediation activities. The Applied Biosystems™ MycoSEQ™ Mycoplasma Detection Kits enable high-sensitivity, sameday, real-time PCR results for early mycoplasma detection, without using live mycoplasma. • Meets or exceeds the recommended regulatory guidance of 10 CFU/mL or 10 genome copies /mL for a variety of mycoplasma species • Highly specific with no cross-reactivity to closely related non-mycoplasma bacterial species • Workflow delivers actionable results in less than 5 hours • Highly accurate, including a discriminatory positive control helps distinguish between true mycoplasma and accidental contamination Residual host-cell DNA quantitation Residual DNA is the genetic material left over from the host cell culture, following mAb purification. The levels of residual DNA in the final product must be as low as possible to meet regulatory guidelines, so accurate testing is required before lot release. The Applied Biosystems™ resDNASEQ™ Residual Host Cell DNA Quantitation Kits are a robust portfolio of real-time and digital PCR assays for a wide variety of host cell lines. • Meets or exceeds recommended regulatory guidance, including test sample LOQ for mammalian DNA and bacteria/yeast DNA • No observed cross-reactivity to unrelated DNA; able to detect fragmented and unfragmented DNA for accurate quantitation • Leverage sample prep protocols to help ensure high precision DNA recovery • Consistent performance kit-to-kit, lot-to-lot and year-to-year For Research Use Only. Not for use in diagnostic procedures. © 2024 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. TaqMan is a trademark of Roche Molecular Systems, Inc., used under permission and license.