Application of affinity capillary electrophoresis for charge heterogeneity profiling of biopharmaceuticals

• Published in: Electrophoresis Vol. 40; no. 22; pp. 3014 - 3022
• Main Authors: Hutanu, Andrei, Kiessig, Steffen, Bathke, Andrea, Ketterer, Rolf, Riner, Sonja, Olaf Stracke, Jan, Wild, Markus, Moritz, Bernd
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 01-11-2019; John Wiley and Sons Inc
• Subjects: Affinity; Affinity capillary electrophoresis; Antibodies, Monoclonal - analysis; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - isolation & purification; Biological Products - analysis; Biological Products - chemistry; Biological Products - isolation & purification; Capillary electrophoresis; Charge heterogeneity testing; Charging; Electric fields; Electrophoresis; Electrophoresis, Capillary - methods; Flow‐through partial filling; Guanidine hydrochloride; Heterogeneity; Ligands; Monoclonal antibodies; Monoclonal antibody co‐formulation; Part V. Proteins, Proteomics and 2D; Quality control; Flow-through partial filling; Affinity capillary electrophoresis; Charge heterogeneity testing; Monoclonal antibody co-formulation
• ISSN: 0173-0835; 1522-2683
• DOI: 10.1002/elps.201900233

Charge heterogeneity profiling is important for the quality control (QC) of biopharmaceuticals. Because of the increasing complexity of these therapeutic entities [1], the development of alternative analytical techniques is needed. In this work, flow‐through partial‐filling affinity capillary electrophoresis (FTPFACE) has been established as a method for the analysis of a mixture of two similar monoclonal antibodies (mAbs). The addition of a specific ligand results in the complexation of one mAb in the co‐formulation, thus changing its migration time in the electric field. This allows the characterization of the charged variants of the non‐shifted mAb without interferences. Adsorption of proteins to the inner capillary wall has been circumvented by rinsing with guanidine hydrochloride before each injection. The presented FTPFACE approach requires only very small amounts of ligands and provides complete comparability with a standard CZE of a single mAb.