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Application of affinity capillary electrophoresis for charge heterogeneity profiling of biopharmaceuticals
Author(s) -
Hutanu Andrei,
Kiessig Steffen,
Bathke Andrea,
Ketterer Rolf,
Riner Sonja,
Olaf Stracke Jan,
Wild Markus,
Moritz Bernd
Publication year - 2019
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201900233
Subject(s) - capillary electrophoresis , chemistry , chromatography , monoclonal antibody , electrophoresis , guanidine , affinity electrophoresis , antibody , affinity chromatography , biochemistry , biology , immunology , enzyme
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.