Identification of a CE-SDS shoulder peak as disulfide-linked fragments from common C_H2 cleavages in IgGs and IgG-like bispecific antibodies.

ABSTRACT: Fragmentation is a well-characterized degradation pathway of therapeutic antibodies and is usually monitored by capillary electrophoresis-sodium dodecyl sulfate (CE-SDS). Although fragments due to cleavage in C_H2 domains linked by intrachain disulfide bonds are common and can be detected by reduced reversed-phase - liquid chromatography mass spectrometry (RP-LCMS) and reduced CE-SDS methods, their separation in nonreduced CE-SDS (nrCE-SDS) has not been reported but speculated as comigrating with intact IgG. A shoulder peak in nrCE-SDS was observed in the stability samples of an IgG-like bispecific antibody and was determined to be mainly caused by fragments from clipping at the C-terminus of leucine (L)306 or L309 (EU numbering) in the C_H2 domain of both heavy chains

SUBMITTER: Cao M 

PROVIDER: S-EPMC8565840 | biostudies-literature | 2021 Jan-Dec

REPOSITORIES: biostudies-literature