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.
Highlights
The biotechnologically engineered monoclonal antibody contains four polypeptides linked via disulfide bonds: two light and two heavy chains
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
A buffer of pH=5.2 with several additives was used to suppress the interactions between the Rtx and the capillary, even an uncoated fused silica capillary was applicable for the efficient separations of several charge variants and the main component
Summary
The biotechnologically engineered monoclonal antibody (mAb) contains four polypeptides linked via disulfide bonds: two light and two heavy chains. It is produced in mammalian cell culture. The mAbs can undergo post-translational modification (deamidation, oxidation, glycosylation, lysine truncation, aggregation) resulting in molecular heterogeneity, charge and size variants. Since these varieties of the mAb have effects on its pharmaceutical properties (antigen binding) and stability, it is important to investigate its molecular heterogeneity. Numerous works were published about the analysis of mAb and the characterization of the molecular heterogeneity [1]. The rituximab (Rtx) is a monoclonal antibody used to treat cancers of blood system such as B cells leukemia (non Hodgkin’s lymphoma) and some autoimmune diseases (rheumatoid arthritis) [2]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: Journal of Chromatography & Separation Techniques
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
