Capillary electrophoresis for rapid identification of monoclonal antibodies for routine application in hospital

Abstract

mAbs are widely used in cancer therapy. Their compounding, performed just before their administration to patients, is executed in a production unit of the hospital. Identification of these drugs, individually prepared in bags for infusion before patient administration, is of paramount importance to detect potential mistakes during compounding stage. A fast and reliable analytical method based on CZE combined to a cationic capillary coating (hexadimethrine bromide) was developed for identification of the most widely used compounded therapeutic for cancer therapy (bevacizumab, cetuximab, rituximab, and trastuzumab). Considering the high structural and physico-chemical similarities of these mAbs, an extensive optimization of the BGE composition has been performed. The addition of perchlorate ions and polysorbate in the BGE greatly increased the resolution. To validate the method, an internal standard was used and the relative migration times (RTm) were estimated. Very satisfactory RSDs of the RTm for rituximab (0.76%), cetuximab (0.46%), bevacizumab (0.31%), and trastuzumab (0.60%) were obtained. The intraday and interday RSD of the method were less than 0.32 and 1.3%, respectively for RTm. Significant differences between theses RTms have been demonstrated allowing mAbs identification. Finally, accurate mAbs identification has been demonstrated by a blind test.

Cutting-edge capillary electrophoresis characterization of monoclonal antibodies and related products

“…They compared their method to conventional SDS-PAGE and concluded on the MZE advantages in terms of faster separation and convenience of handling the microfluidics-based technology. Finally, Jacoulet et al proposed a fast and reliable CZE method combined to a cationic capillary coating to identify the most widely used mAbs for cancer therapy[86].…”

High‐resolution separation of monoclonal antibodies mixtures and their charge variants by an alternative and generic CZE method

“…Jaccoulet et al . proposed a denaturing CZE method involving the use of a BGE buffered at pH 3.0 for the separation of bevacizumab, cetuximab, trastuzumab and rituximab within 15 min.…”

“…To ensure patients safety, the confirmation of the mAb identity should be performed to prevent potential mistake at the compounding stage. Jaccoulet et al [27] proposed a denaturing CZE method involving the use of a BGE buffered at pH 3.0 for the separation of bevacizumab, cetuximab, trastuzumab and rituximab within 15 min. In addition to the large separation of the main species of bevacizumab, trastuzumab and rituximab within 45 min, our methodology also successfully separated the charge variants of the three mAbs.…”

Recent advances in protein analysis by capillary and microchip electrophoresis

“…149 A CZE method with cationic capillary coating for the identification of mAb in routine mAb for cancer therapy in hospitals was reported. 154 Characterization of ADC was performed by sheathless CE-MS/MS with nanoESI infusion for detection of species with different number of conjugated drugs has also been described. 155 A characterization of therapeutic antibodies by CE-ESI-MS allows for rapid sequencing and degradation hotspot analysis, and this work achieved enhanced sensitivity for poorly ionizing molecular species like glycoproteins.…”