Heightened Characterization of Therapeutic mAbs Using a Novel Enzyme-Facilitated Middle-Down Approach

• Broadcast Date: Wednesday, May 15, 2013
• Time: 1 pm ET, 10 am PT

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Quality control and analysis of monoclonal antibody (mAb) therapeutics is required throughout therapeutic development and production. In particular, deep characterization of glycosylation and other post-translational modifications is required because of their influence on antibody stability, activity, and function. The complexity of cell culture systems used to produce these proteins further complicates analyses. The majority of analytical methodologies are time consuming and relatively labor intensive.

In this webinar, the speakers will introduce the Antibody Subunit/Domain Mapping assay, a new characterization method enabled using ultrahigh-resolution mass spectrometry and a novel IgG-specific enzyme, FabRICATOR (IdeS). In contrast to limited proteolysis using Lys-C or Papain, the FabRICATOR enzyme enables rapid, single site-specific cleavage of an IgG thus resulting in three distinct mAb fragments after further processing. The three fragments range in size between 22–26 kilodaltons and are more amenable to further MS analysis compared to intact mAb molecules. The enzyme is very robust, efficient, and facilitates complete sample digestion in less than half an hour for individual antibody candidates. This characterization approach has be applied to molecules other than native IgGs such as ADCs, bi-specific IgGs, and Fc fusion proteins.

Speakers will also detail how the use of the FabRICATOR enzyme facilitates the characterization of multiple post-translational modifications (i.e., N-glycosylation, methionine oxidation, etc.) along with demonstrating the rapid confirmation of the amino acid sequence of the therapeutic protein within a few hours. This approach reduces the number of fragments generated by conventional enzymes, limits digestion variability, and minimizes the generation of method-induced degradation products commonly observed in bottom-up characterization approaches for protein therapeutics.

What You Will Learn

• How these novel enzyme can rapidly generate specific antibody fragments for use in bioanalytics
• How the generation of a smaller number of proteolytic products in the middle-down approach simplifies LC-MS analysis
• How this enzyme can be used for rapid comparability studies of biosimilars vs. innovator compounds
• How these enzyme systems can be used to screen production clones, therapeutic antibody molecules
• How antibody glycoprofiling and post-translational modification analysis can be more rapidly characterized

Who Should Attend

• Therapeutic antibody developers
• Recombinant antibody QC/QA scientists
• Antibody production scientists
• Protein biochemists
• Protein/biological protein mass spectroscopists
• Carbohydrate chemists
• Glycoprotein chemists

A live Q&A session will follow the presentations, offering you a chance to pose questions to our expert panelists.

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