A Single Weak Cation Exchange Liquid Chromatographic Method ((WCX)HPLC/DAD) for the of Study of Five Marketed Therapeutic Monoclonal Antibodies

Abstract

Marketed monoclonal antibodies (mAbs) represent a very important class of the current biopharmaceutics. The great complexity of their structure made necessary the use of different analytical approaches for assessing different physico-chemical properties. In this work, weak cation exchange (CEX) high performance liquid chromatography with diode array detection ((CEX)HPLC/DAD) is used to assess the charge variant profile. The method developed combined the effect of ionic strength and controlled pH gradient and allows for the charge variants analysis of the five mAbs studied, namely bevacizumab (BVZ), cetuximab (CTX) infliximab (INF), rituximab (RTX) and trastuzumab (TTZ), which are among the most used mAbs worlwide. The differences in the charge variant in the natural isoforms of the mAbs generated characteristic CEX chromatograms for each of mAbs that can be useful for identification purposes. These chromatograms have provided to be very useful to track changes in the charge variants of each particular mAb both in controlled degraded studies and in long-term stability study of in-use samples solutions at 2 mg/mL in 0.9% NaCl stored refrigerated (at 4 oC) and frozen (-20 oC) for two months.