Applications of capillary electrophoresis for biopharmaceutical product characterization

Abstract

Capillary electrophoresis (CE), after being introduced several decades ago, has carved out a niche for itself in the field of analytical characterization of biopharmaceutical products. It does not only offer fast separation, high resolution in miniaturized format, but equally importantly represents an orthogonal separation mechanism to high‐performance liquid chromatography. Therefore, it is not surprising that CE‐based methods can be found in all major pharmacopoeias and are recommended for the analysis of biopharmaceutical products during process development, characterization, quality control, and release testing. Different separation formats of CE, such as capillary gel electrophoresis, capillary isoelectric focusing, and capillary zone electrophoresis are widely used for size and charge heterogeneity characterization as well as purity and stability testing of therapeutic proteins. Hyphenation of CE with MS is emerging as a promising bioanalytical tool to assess the primary structure of therapeutic proteins along with any impurities. In this review, we confer the latest developments in capillary electrophoresis, used for the characterization of critical quality attributes of biopharmaceutical products covering the past 6 years (2015–2021). Monoclonal antibodies, due to their significant share in the market, have been given prioritized coverage.

Approaches for the detection and analysis of antidrug antibodies to biopharmaceuticals: A review

“…Several modes of CE have been used to examine antibody‐based drugs and antibody‐drug conjugates. These modes have included CZE, cIEF, and CGE [95, 98].…”

“…CE is a separation technique that is based on the differential migration of analytes through a capillary and in the presence of an applied potential [95][96][97][98]. Factors that affect the rate of migration and mobility of an analyte in CE include the size and charge of the analyte as well as the pH, temperature, and composition of the BGE within the capillary.…”

“…Several modes of CE have been used to examine antibody-based drugs and antibody-drug conjugates. These modes have included CZE, cIEF, and CGE [95,98]. CE has been used to screen ADAs against antibodyrelated biotherapeutics, including nanobodies [96].…”

Global intercompany assessment of ICIEF platform comparability for the characterization of therapeutic proteins

“…Charge heterogeneity is deemed a critical product quality attribute by regulatory agencies and must be evaluated due to the potential impact of acidic and basic isoforms on pharmacokinetics, biological activity, and long-term storage [1,2]. Charge heterogeneity can be monitored by a wide range of analytical methods that are based on LC [3] or electrophoresis [4]. Among the electrophoretic-based methods, CIEF has become the most common method used in the biopharmaceutical industry to characterize charge heterogeneity [4,5].…”

“…Charge heterogeneity can be monitored by a wide range of analytical methods that are based on LC [3] or electrophoresis [4]. Among the electrophoretic-based methods, CIEF has become the most common method used in the biopharmaceutical industry to characterize charge heterogeneity [4,5].…”