N-glycan Profiling of monoclonal Antibody (mAb) on Nexera Bio UHPLC Coupled with Fluorescence Detector and Q-TOF Mass Spectrometer

Introduction

N -linked glycosylation on Asn residue with consensus sequence Asn-X-Ser/Thr (where X is any amino acid except Pro) plays a critical role in stability, bioactivity, and immunogenicity of monoclonal antibodies (mAbs). The N -glycan moieties of therapeutic mAbs, especially biosimilar products, must be adequately and routinely characterized to ensure product quality. In this report, we established a robust, sensitive, and reproducible analytical platform that contains a Nexera Bio UHPLC system, a fluorescence detector (RF-20A), and a Q- TOF mass spectrometer (LCMS-9030) for N -glycan profiling of bevacizumab biosimilar. N -glycans were released from bevacizumab biosimilar with PNGase F, labeled with 2-aminobenzamide (2-AB), and subsequently detected via RF-20A and LCMS-9030. LCMS-9030 was applied for peak assignment using an accurate mass of corresponding N -glycans, while peak areas from RF-20A were used for N -glycan quantitation.

December 12, 2019 GMT