Biophysical Characterization Of A Therapeutic mAb And Its Associated Antigen-Binding Fragments

Changes during purification, formulation, fill-finish, and distribution can affect protein stability and practically all steps in the manufacturing process may perturb the higher-order structure and heterogeneity of the molecule, thus determining size homogeneity of a monoclonal antibody in solution is important for comparability and characterization of these medications. For this study, an IgG antibody was cleaved into antigen-binding fragments using pepsin and papain digestion, followed by protein purification. The fragments, along with intact mAb, were studied by Sedimentation Velocity-Analytical Ultracentrifugation (SV-AUC) and Size-Exclusion Chromatography Multi-Angle Light Scattering (SEC-MALS) to determine sedimentation coefficient, distribution, and molecular weight of each.

Download the poster to learn more about the results from the study and characterization of size and distribution of the intact mAb and enzymatic fragments.

Key Learning

• The efficacy of orthogonal approaches, SV-AUC and SEC-MALS
• Effective characterization of intact antibodies, as well as antibody fragments
• Overlapping and complementary results provide ideal context for further aggregation studies