Imaged Capillary Isoelectric Focusing for Charge-Variant Analysis of Biopharmaceuticals
نویسندگان
چکیده
level: interMediAte Analyzing charge variants of therapeutic proteins is critical for characterizing and monitoring quality attributes of antibodies. Charge variants include deamidation, formation of N-terminal pyroglutamate, aggregation, isomerization, sialylated glycans, antibody fragmentation, and glycation at the lysine residues. In some cases, such changes affect binding, biological activity, patient safety, and shelf life. The biopharmaceutical industry relies on tools such as ionexchange chromatography (IEC), isoelectric-focusing gel electrophoresis (IEF), and capillary equivalents such as capillary isoelectric focusing (CIEF) and imaged CIEF (iCIEF) to characterize charge variants. iCIEF has contributed significantly to biopharmaceutical development with its high resolution, minimal development time, reduced sample volume, and fast run times (1–3). Those benefits allow for applications across entire pharmaceutical processes, from cell culture development and optimization to commercial quality control (QC) release and stability activities. Here, we describe its application in QC and characterization (product and process). We focus on unique advantages for analyzing drug– antibody conjugates, and provide an in-depth example of iCIEF’s power for evaluating process changes. Finally, we detail a novel application for iCIEF profile characterization by mass spectrometry.
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