Di Marco, Fiammetta, Berger, Thomas, Esser-Skala, Wolfgang, Rapp, Erdmann, Regl, Christof and Huber, Christian (2021) Simultaneous Monitoring of Monoclonal Antibody Variants by Strong Cation-Exchange Chromatography Hyphenated to Mass Spectrometry to Assess Quality Attributes of Rituximab-Based Biotherapeutics. International journal of molecular sciences, 22 (16). ISSN 1422-0067

Abstract

Different manufacturing processes and storage conditions of biotherapeutics can lead to a significant variability in drug products arising from chemical and enzymatic post-translational modifications (PTMs), resulting in the co-existence of a plethora of proteoforms with different physicochemical properties. To unravel the heterogeneity of these proteoforms, novel approaches employing strong cation-exchange (SCX) high-performance liquid chromatography (HPLC) hyphenated to mass spectrometry (MS) using a pH gradient of volatile salts have been developed in recent years. Here, we apply an established SCX-HPLC-MS method to characterize and compare two rituximab-based biotherapeutics, the originator MabThera® and its Indian copy product Reditux™. The study assessed molecular differences between the two drug products in terms of C-terminal lysine variants, glycosylation patterns, and other basic and acidic variants. Overall, MabThera® and Reditux™ displayed differences at the molecular level. MabThera® showed a higher degree of galactosylated and sialylated glycoforms, while Reditux™ showed increased levels of oligomannose and afucosylated glycoforms. Moreover, the two drug products showed differences in terms of basic variants such as C-terminal lysine and N-terminal truncation, present in Reditux™ but not in MabThera®. This study demonstrates the capability of this fast SCX-HPLC-MS approach to compare different drug products and simultaneously assess some of their quality attributes.

Item Type:	Article
Keywords:	Antibodies, Monoclonal Antineoplastic Agents, Immunological Biosimilar Pharmaceuticals Cations Chromatography, Affinity Chromatography, High Pressure Liquid Glycosylation Mass Spectrometry Rituximab
Date Deposited:	23 Jun 2022 00:45
Last Modified:	23 Jun 2022 00:45
URI:	https://oak.novartis.com/id/eprint/45081