Techniques & Tools Pharma & Biopharma, Mass Spectrometry

Rapid and Robust Subunit Domain Mapping of Monoclonal Antibody Based Biotherapeutics

Highly specific enzymes for middle down approach - rapid mass spectrometric analysis of antibody based biotherapeutics using FabRICATOR®

05/27/2014

Digestion with the FabRICATOR^® (IdeS) enzyme, which specifically cleaves IgG under the hinge region, followed by reduction results in 3 distinct mAb subunits (LC, Fd and Fc/2) in the range of 22-27 kDa. Optionally, selective Fc-deglycosylation can be performed using IgGZERO™ (EndoS) or GlycINATOR™ (EndoS2) to further reduce heterogeneity. Either of the two enzymes can be co-incubated with FabRICATOR ® during the digestion step. They facilitate characterization of posttranslational modifications like glycan heterogeneity, oxidation, lysine clipping and a rapid confirmation of amino acid sequence. The method can be applied for tasks like clone selection, process development, formulation and stability studies of monoclonal antibody based bio therapeutics such as MAb’s, Fc-fusion proteins, biosimilars and ADC’s.

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