Unambiguous identification and quantification of galactose-α-1,3-galactose as a critical quality attribute with mass spectrometry and exoglycosidases.

ABSTRACT: Alpha-galactosylation, galactose-α-1,3-galactose (α-Gal), is always regarded as a critical quality attribute due to its potential to provoke immunogenic responses in patients. Consequently, monitoring alpha-galactosylation in therapeutic proteins is essential, but current analytical techniques fall short in terms of identification sensitivity and quantification accuracy. Specifically, the released glycan assay by hydrophilic interaction liquid chromatography-fluorescence-mass spectrometry, the gold standard for glycan separation/identification, faces challenges due to ambiguities with isomeric glycan structures. To address these challenges, we developed a comprehensive analytical method that enhances both the identification sensitivity and quantification accuracy for α-Gal. We developed an

SUBMITTER: Gan Y 

PROVIDER: S-EPMC12622356 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature