Identification of critical chemical modifications by size exclusion chromatography of stressed antibody-target complexes with competitive binding.

ABSTRACT: Chemical modifications (attributes) in the binding regions of stressed therapeutic proteins may affect binding to target and efficacy of therapeutic proteins. The method presented here describes the criticality assessment of therapeutic antibody modifications by size-exclusion chromatography (SEC) of competitive binding between a stressed antibody and its target, human epidermal growth factor receptor-2 (HER2), followed by SEC fractionation and peptide mapping characterization of bound and unbound antibodies. When stressed antibody and its target were mixed at a stoichiometric molar ratio of 1:2, only antibody-receptor complex eluted from SEC, indicating that binding was not decreased to break the complex. When a smaller amount of the receptor was provided (1:1), the antibody species with

SUBMITTER: Shi RL 

PROVIDER: S-EPMC7899689 | biostudies-literature | 2021 Jan-Dec

REPOSITORIES: biostudies-literature