AUTHOR=Kim Hyejin , Bang Geul , Park Ye Eun , Park Moonhee , Choi Jung Hoon , Oh Myung Jin , An Hyun Joo , Yoo Jong Shin , Park Youngja Hwang , Kim Jin Young , Hwang Heeyoun TITLE=Advanced assessment through intact glycopeptide analysis of Infliximab’s biologics and biosimilar JOURNAL=Frontiers in Molecular Biosciences VOLUME=9 YEAR=2022 URL=https://www.frontiersin.org/journals/molecular-biosciences/articles/10.3389/fmolb.2022.1006866 DOI=10.3389/fmolb.2022.1006866 ISSN=2296-889X ABSTRACT=

Characterization of therapeutic monoclonal antibodies (mAbs) represents a major challenge for analytical sciences due to their heterogeneity associated with post-translational modifications (PTMs). The protein glycosylation requires comprehensive identification, which could influence on the mAbs’ structure and their function. Here, we demonstrated high-resolution tandem mass spectrometry with an ultra-high-performance liquid chromatography for characterization and comparison between biologics and biosimilar of infliximab at an advanced level. Comparing the N- and O-glycopeptides profiles, a total of 49 and 54 glycopeptides was identified for each product of the biologics and biosimilar, respectively. We also discovered one novel N-glycosylation site at the light chain from both biopharmaceuticals and one novel O-glycopeptide at the heavy chain from only biosimilar. Site-specific glycopeptide analysis process will be a robust and useful technique for evaluating therapeutic mAbs and complex glycoprotein products.