Method validation of Capillary Electrophoresis–Sodium Dodecyl Sulfate for Identity and Purity analysis of Monoclonal Antibodies

Abstract

       Capillary electrophoresis–sodium dodecyl sulfate (CE-SDS) is a qualitative technique for protein analysis and is widely use for separation of protein by molecular size. This technique is increasingly applied to analysis of biotherapeutic products derived from biotechnology including monoclonal antibodies (mAbs). This study, validation of CE-SDS technique for identity and purity analysis of biotherapeutic products was performed by using Beckman PA 800, capillary bare-fused silica, SDS-MW Gel buffer with photodiode array (PDA) detector at wavelength 220 nm. The results showed that it has high resolution for detection of product – related impurity such as non-glycosylated heavy chain: NGHC. The obtained data has specificity and precision related to standard criteria as shown that the %RSD of migration time (MT) and corrected percentage area of light chain: LC, heavy chain: HC and NGHC were less than 3. The robustness was also determined by using new lot of SDS gel buffer and capillary in analysis. It’s found that only %RSD of MT of HC and NGHC were slightly exceed 3 when using new capillary. This method was used to analyze 2 mAb samples, the results were correlated with results of standard material supported that CE-SDS is suitable technique for using as standard method for identity and purity analysis for quality control of marketed mAbs in Thailand.