Analytical comparability study of anti-CD20 monoclonal antibodies rituximab and obinutuzumab using a stability-indicating orthogonal testing protocol: Effect of structural optimization and glycoengineering.

This study focuses on analytical comparability study of anti-cd20 monoclonal antibodies rituximab and obinutuzumab using a stability-indicating orthogonal testing protocol: effect of structural optimization and glycoengineering.. The research employs high-performance liquid chromatography (HPLC) techniques to address analytical challenges in the biopharmaceutical field. Glycoengineering and biosimilarity are the key factors for growing, promising and progressive approaches in monoclonal antibodies development. In this study, the physicochemical stability of anti-CD20 rituximab (RTX); originator and biosimilar was compared to its glycoengineered humanized version; obinutuzumab (OBZ). An orthogonal stability-indicating protocol using a set of validated bioanalytical techniques;... Research Background and Significance The analytical comparability of monoclonal antibodies (mAbs) is a critical step in biopharmaceutical development, especially when evaluating originator biologics against biosimilars or glycoengineered variants. Rituximab (RTX) and obinutuzumab (OBZ) are anti-CD20 mAbs widely used for targeting B-cell malignancies. While RTX represents a chimeric originator antibody, OBZ is a humanized glycoengineered antibody designed to enhance therapeutic efficacy and reduce immunogenicity. This study is significant as it addresses the complex analytical challenges in characterizing these structurally related but functionally distinct antibodies. It underscores the importance of stability-indicating orthogonal testing protocols, integrating multiple validated bioanalytical techniques including HPLC, to assess physicochemical stability and comparability. The insights gained are essential for ensuring product quality, safety, and efficacy, supporting regulatory approval and g