The Impact of Immunoglobulin G1 Fc Sialylation on Backbone Amide H/D Exchange.

ABSTRACT: The usefulness of higher-order structural information provided by hydrogen/deuterium exchange-mass spectrometry (H/DX-MS) for the structural impact analyses of chemical and post-translational antibody modifications has been demonstrated in various studies. However, the structure-function assessment for protein drugs in biopharmaceutical research and development is often impeded by the relatively low-abundance (below 5%) of critical quality attributes or by overlapping effects of modifications, such as glycosylation, with chemical amino acid modifications; e.g., oxidation or deamidation. We present results demonstrating the applicability of the H/DX-MS technique to monitor conformational changes of specific Fc glycosylation variants produced by in vitro glyco-engineering technology. A trend towards less H/DX in Fc C?2 domain segments correlating with larger glycan structures could be confirmed. Furthermore, significant deuterium uptake differences and corresponding binding properties to Fc receptors (as monitored by SPR) between ?-2,3- and ?-2,6-sialylated Fc glycosylation variants were verified at sensitive levels.

SUBMITTER: Kuhne F 

PROVIDER: S-EPMC6963987 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature