Enzymatic Desialylation Enables Reliable Charge Variant Characterization of Highly Glycosylated and Sialylated Fc Fusion Proteins.

ABSTRACT: Fusion proteins constitute a class of engineered therapeutics and have emerged as promising candidates for disease treatment. However, the structural complexity and heterogeneity of fusion proteins make their characterization extremely challenging, and thus, an innovative and comprehensive analytical toolbox is needed. Here, for the first time, we demonstrate a novel and robust workflow to evaluate charge variants for a highly glycosylated fusion protein with heavy sialylation using imaged capillary isoelectric focusing (icIEF). In the development of the icIEF method, key factors that were systematically investigated include the desialylation level, the stability of the desialylated molecule, incubation time and temperature of desialylation, protein concentrations, urea and l-arginine effe

SUBMITTER: Wen X 

PROVIDER: S-EPMC11833721 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature