Accelerated development of a SEC-HPLC procedure for purity analysis of monoclonal antibodies using design of experiments.

This study focuses on accelerated development of a sec-hplc procedure for purity analysis of monoclonal antibodies using design of experiments.. The research employs high-performance liquid chromatography (HPLC) techniques to address analytical challenges in the biopharmaceutical field. The complex structure of biopharmaceutical products poses an inherent need for their thorough characterization to ensure product quality, safety, and efficacy. Analytical size exclusion chromatography (SEC) is a widely used technique throughout the development and manufacturing of monoclonal antibodies (mAbs) which quantifies product size variants such as aggregates and fragments.... Research Background and Significance Monoclonal antibodies (mAbs) have become cornerstone biopharmaceutical agents, extensively utilized for treating a wide range of diseases including cancer, autoimmune disorders, and infectious diseases. Their complex molecular architecture, comprising multiple domains and potential post-translational modifications, presents significant analytical challenges, especially in purity and size variant characterization. Size exclusion chromatography (SEC) coupled with high-performance liquid chromatography (HPLC) is a pivotal technique in this context, routinely applied for the quantitation of aggregates, fragments, and other size-related impurities. The accurate assessment of these variants is critical for ensuring therapeutic efficacy, safety, and regulatory compliance. This study addresses the growing demand for accelerated and robust SEC-HPLC methods, leveraging design of experiments (DoE) approaches to optimize chromatographic conditions efficiently. By systematically exploring the interplay of chromatographic parameters, the research aims to establish a validated, reliable