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

This study focuses on comparison of originator and biosimilar monoclonal antibodies using hrms, fc affinity chromatography, and 2d-hplc.. The research employs high-performance liquid chromatography (HPLC) techniques to address analytical challenges in the biopharmaceutical field. 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... Research Background and Significance Monoclonal antibodies (mAbs) represent a cornerstone in modern biopharmaceutical therapeutics due to their specificity and efficacy in treating a variety of diseases, including cancers and autoimmune disorders. The complexity of these biologics arises from their large molecular size, post-translational modifications, and intricate tertiary structures, which pose substantial analytical challenges. Because of the inherent complexity, biosimilars—follow-on versions of originator mAbs—cannot be exact replicas but must be shown to be highly similar in structure and function to the originator to ensure comparable safety and efficacy. This study addresses the critical need for robust analytical methods capable of differentiating subtle molecular differences between originator and biosimilar mAbs. The combined use of high-resolution mass spectrometry (HRMS), Fc affinity chromatography, and advanced two-dimensional HPLC (2D-HPLC) provides a comprehensive characterization platform. Such multidimensional analytical strategies are vital for regulatory approval processes and quality control in biopharmaceutical dev