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Simple, Accurate Large Molecule Bioanalysis
Industry Insight

Simple, Accurate Large Molecule Bioanalysis

Simple, Accurate Large Molecule Bioanalysis
Industry Insight

Simple, Accurate Large Molecule Bioanalysis

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Biopharmaceutical, translational, clinical and sports anti-doping labs performing high-throughput characterization of large molecules are commonly challenged by the limitations of conventional affinity purification methods. These challenges include low efficiency, inconsistent results, high background and carryover and the inability to standardize methods and transfer between labs. 

To discuss these challenges and how Thermo Scientific are helping  to overcome them we spoke to Preeti Saini, product manager for MSIA liquid handling consumables at Thermo. 

What are some common limitations of traditional affinity purification methods and how will the Thermo Scientific™ MSIA™ microcolumns for Tecan’s Freedom EVO® platform help to overcome these?

Traditional affinity purification methods have several challenges including insufficient sensitivity at low levels of detection (picogram quantities); low efficiency, high background noise and carryover, sub-standard data quality and reproducibility, and the inability to standardize and transfer analytical methods between labs. The Thermo Scientific MSIA microcolumns for the Freedom EVO series overcomes these challenges by combining the robust automated high-throughput liquid handling capabilities of the Tecan Freedom EVO series of instruments with the novel affinity purification capabilities of MSIA technology. The proprietary MSIA microcolumns are densely coated with a target-specific affinity ligand for effective and efficient analytical-grade affinity purification of even low-abundance target analytes.

Housed within a pipette tip, these microcolumns enable users to affinity purify the target analyte from complex biological matrices with greater ease and simplicity as compared to other affinity purification technologies on the market. This unique affinity capture method allows the detection and simultaneous qualitative and quantitative analyses of ≥ pg/mL levels of intact or digested proteins without the use of mass tags or labeling reagents. 
By combining MSIA technology with Tecan’s Freedom EVO platform, what application needs does the new workflow address?

MSIA Streptavidin EVO microcolumns can benefit many applications, including:

• Pre-clinical and clinical trials of biotherapeutics
• Therapeutic antibody analysis
• Biotransformation [Drug antibody ratio (DAR), and drug antibody conjugate (DAC)]
• Analysis of protein variants and post-translational modifications
• Biomarker discovery and analysis
• Detection and characterization of exogenous biomolecules in sports doping

In fact, most applications requiring qualitative or quantitative information from large molecules such as protein, peptides and antibodies can benefit from this technology. Laboratories looking to maximize efficiency when conducting high-throughput large molecule analyses can also take advantage of the automation capabilities of the Tecan EVO platform to streamline automated workflows.

In terms of efficiency, what new benefits does the technology provide to researchers performing large molecule bioanalysis?

Researchers performing large molecule bioanalysis can use this technology for faster assay development, and to preserve their precious samples – even 10 µL is sufficient. Compared to traditional affinity purification methods, the MSIA Streptavidin-EVO method involves a shorter incubation time, and unlike bead-based technologies, there is no need to aliquot samples.

The Freedom EVO platform boosts productivity by allowing parallel processing of 96 samples at a time, while improving consistency and reproducibility. The MSIA workflow produces minimal analytical variability and improved signal-to-noise ratios, reducing the likelihood that experiments will need to be repeated. Unlike resin-based supports, the microcolumns are resistant to compression and there is no affinity matrix loss, increasing LC-MS instrument lifespan and making it a cost-effective solution.

The MSIA technology is available in a range of formats to support individual assay development needs. What are these and what application areas do they cover?

There are several different formats of MSIA technology available, in order to meet the needs of different users and applications. 

• The MSIA Streptavidin EVO microcolumns employ streptavidin to bind biotin-conjugates and are designed specifically for use with the Tecan Freedom EVO instrument for high throughput large molecule analysis. 

• The Thermo Scientific™ MSIA D.A.R.T.’s™ (Disposable Automated Research Tips) are compatible with both the Thermo Scientific™ Versette™ automated liquid handler and the Thermo Scientific™ semi-automated Finnipipette™ Novus i multichannel electronic pipette, making them ideal for high- and low-throughput analyses. 

o MSIA D.A.R.T.’s are available in Protein A, G and combined A and G; streptavidin; and insulin formats. Protein A, G and A/G provides a versatile option to bind and orientate a wide range of antibodies, enabling customer flexibility to use antibodies of their choice specific to their applications 

o MSIA Insulin D.A.R.T.’s employ an antibody against a conserved region in the insulin peptide for the simultaneous quantitative and qualitative bioanalysis of insulin, including endogenous and therapeutic analogs such as Lantus™, Apidra™ etc. 

o If a customer has an analyte of interest that cannot be captured by these products, we offer custom MSIA D.A.R.T.’s to tailor specific requests.

Preeti Saini was speaking to Jack Rudd, Editor at Technology Networks.