Computational insights into O-glycosylation in a CTLA4 Fc-fusion protein linker and its impact on protein quality attributes.

ABSTRACT: The hinge region of immunoglobulin G1 (IgG1) is used as a common linker for Fc-fusion therapeutic proteins. With the advances of high-resolution mass spectrometry and sample treatment strategies, unexpected O-linked glycosylation has been observed in the linker. However, the molecular mechanism involved in this unusual posttranslational modification is unknown. In this study, we applied site-direct mutagenesis, mass spectrometry, analytical chromatography, and computational modeling to investigate O-glycosylation processes in a clinically used CTLA4 Fc-fusion protein and its impacts on protein quality attributes. Surprisingly, O-glycans could be formed at new sites when an initial O-glycosylation site was eliminated, and continued to occur until all potential O-glycosylation sites were nul

SUBMITTER: Song Y 

PROVIDER: S-EPMC7734232 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature