earticle

영어

Bioactivity and efficacy of therapeutic protein are dependent on structure-specific glycan attached to the therapeutic proteins. Therefore, precise glycomic characterization and monitoring are essential in the manufacture of therapeutic glycoproteins. However, the inherent structural diversity of glycosylation significantly hinders analysis. Thus, it is challenging to comprehensively characterize structure–specific glycans. We propose isomer-specific LC/MS and MS/MS screening as a method for rapid identification and structural elucidation of biopharmaceutical glycosylation. Biotherapeutic glycoproteins including mAbs, and EPOs were prepared and analyzed. After removing salts and detergents, N-glycans were enzymatically released and purified by graphitized carbon solid phase extraction. PGC chip nano-LC/Q-TOF provided chromatographic isomer separation profiling and structural characterization. Multiple structure isomers including antennary, polylactosamine, alpha galactose epitope, and linkage iosomers were identified within isomeric compositions. A database of accurate mass, RT, and diagnostic MS/MS fragments was developed and applied to identify and structurally characterize biopharmaceutical glycans. The approach is fast, does not need for labeling or derivatisation and is applicable for highly accurate identification of biotherapeutic glycosylation.