Bottom-up characterization of a monoclonal antibody Trastuzumab with sheathless CESI-MS coupled to the Orbitraps mass spectrometers

摘要

Trastuzumab is a humanized monoclonal antibody used to treat metastatic breast cancer. In the US, the patent on Trastuzumab expires in 2014, which opens up the opportunity for worldwide biosimilar competition. The current FDA guidelines for approval of biosimilars require rigorous characterization of candidate heterogeneity, including glycoforms and degradation products, whose analysis relies heavily on mass spectrometry (MS). Capillary electrophoresis (CE) is well suited to the separation of such species due to their differences in charge states and Stoke's radii. Recent advances in electrospray ion source technology have allowed the sheathless coupling of CE to MS, eliminating the dilution effects of prior sheathflow techniques, potentially allowing complete sequence coverage and glycoform characterization in single runs with reduced sample manipulation.