Comparison of originator and biosimilar monoclonal antibodies using HRMS, Fc affinity chromatography, and 2D-HPLC

• Published in: Analytical and bioanalytical chemistry Vol. 414; no. 23; pp. 6761 - 6769
• Main Authors: Reinders, Lars M. H., Klassen, Martin D., Teutenberg, Thorsten, Jaeger, Martin, Schmidt, Torsten C.
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01-09-2022; Springer; Springer Nature B.V
• Subjects: Affinity; Affinity chromatography; Analytical Chemistry; Analytical methods; Biochemistry; Biological products; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Chromatography; Comparative analysis; Fc receptors; Food Science; Glycan; High performance liquid chromatography; High resolution; Laboratory Medicine; Liquid chromatography; Manufacturing industry; Mass spectrometry; Mass spectroscopy; Mathematical analysis; Monitoring/Environmental Analysis; Monoclonal antibodies; Orthogonality; Peptide mapping; Peptides; Research Paper; Rituximab; Scientific imaging; Spectroscopy; Structure-function relationships; Trastuzumab; Two dimensional analysis; Germany; High-resolution mass spectrometry (HRMS); Monoclonal antibody (mAb); FcR affinity chromatography; Biosimilar; 2D-HPLC
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-022-04236-8

Due to the complex manufacturing process of therapeutic monoclonal antibodies, it is hardly possible to produce an identical copy of the original product (originator). Consequently, follow-on products (biosimilars) must demonstrate their efficacy being similar to the originator in terms of structure and function. During this process, a variety of analytical methods are required for this purpose. This study focuses on three particularly relevant analytical techniques: high-resolution mass spectrometry, fragment crystallisable (Fc) affinity chromatography, and two-dimensional peptide mapping. Each analytical method proved able to identify specific differences between originator and biosimilar. High-resolution mass spectrometry was used to characterize the glycan pattern. It was shown that a trastuzumab biosimilar did not have the G0:G0F sugar modification identified in the originator. The application of affinity chromatography to rituximab showed that originator and biosimilar interacted differently with the immobilized Fc receptor. Furthermore, 2D-HPLC peptide mapping demonstrated the influence of orthogonality of separation dimensions, leading to differentiation of a rituximab originator and biosimilar.
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