Characterization of Charge Variants in Biotherapeutics

This technical report outlines the use of orthogonal technologies such as capillary isoelectric focusing (cIEF), imaged cIEF (icIEF)-UV/MS, and LC-MS based peptide mapping for the characterization of charge variants in biotherapeutics. The document details how these workflows can be utilized separately or in combination to analyze critical quality attributes (CQAs) throughout the biotherapeutic development process. It emphasizes the importance of charge heterogeneity as a CQA that must be closely monitored to ensure the safety and efficacy of biotherapeutic products. The report also addresses challenges in identifying proteoforms within charge variant peaks and describes how the integration of cIEF, icIEF-UV/MS, and LC-MS techniques can provide comprehensive insights into the charge profile of infliximab. Additionally, it discusses the methodologies for sample preparation, data acquisition, and analysis, highlighting the capabilities of the BioPhase 8800 and ZenoTOF 7600 systems in achieving accurate and rapid characterization of biotherapeutics.