Analysis of Rituximab, A Therapeutic Monoclonal Antibody by Capillary Zone Electrophoresis

Abstract

This paper focuses on the applicability of capillary zone electrophoresis (CZE) using uncoated fused silica capillaries for the determination of heterogeneity of rituximab (MabThera, Roche) and for the study of its solution and thermal stability. The best resolution for the charge variants of the main component was obtained with a buffer electrolyte containing 800 mM 6-amino caproic acid, 2 mM triethylene tetramine and 0.05% hydroxypropyl methylcellulose at pH = 5.2. It was found that the pH and the components of the buffer used for the electrophoretic separation and for the dilution of the sample prior to the analysis are important to the stability of the rituximab. We demonstrated the rituximab is stable in the pharmaceutical product MabThera due to the stabilizing additives, but the dilution of the MabThera caused a slow formation of acidic variants, while the amount of the basic variants did not change. After incubation of the diluted rituximab at higher temperature several charge variants could be determined by CZE.

High‐throughput capillary electrophoresis analysis of biopharmaceuticals utilizing sequential injections

“…Moreover, the presented method can be adapted to perform any separation where acquisition time is considerably longer than the separation window. For example, researchers have reported CE‐UV analysis of rituximab (similar mAb used in the present study) within 10 min with a CE dead time of 9 min [25]. Because the separation window is only about 1 min, at least four‐to‐five samples can easily be analysed in the 10 min total time.…”

Recent advances in capillary electrophoretic migration techniques for pharmaceutical analysis (2013–2015)

“…For example, a repeatability of peak areas of approximately 1% RSD has been obtained. The transferability of this robust method has been demonstrated in an intercompany study with 11 participating labs, and another similar method has proven useful for a stability indicating assay of the mAb Rituximab [200]. As described earlier [1,191], CZE has been used for stability indicating methods, e.g.…”

“…Size variations are now routinely elucidated using CE employing sodium dodecyl sulfate as reagent ("CE-SDS"), similar to SDS-PAGE but superior in terms of validation properties [200,[204][205][206][207]. The related general concepts have been already described, e.g.…”