Mass spectrometry (MS) mass resolution and accuracy, sample complexity remains a challenge when analyzing intact proteins. Separation before MS analysis is needed to achieve reliable and sensitive characterization of minor protein variants and modifications. Integrating capillary electrophoresis separation with electrospray ionization (CESI) offers the selectivity and sensitivity required to distinguish closely related intact protein variants and isoforms by mass spectrometric detection.
Examples will focus on the intact and middle-up analysis of proteins of pharmaceutical and biomedical interest. These include monoclonal antibodies (mAbs), single-domain antibodies (nanobodies) targeting GPCR receptors and amyloidogenic protein beta-2-microglobulin.
Key Learning Points
Learn about the analysis of protein conformational changes under native conditions.
Understand how CESI-MS was used to reveal detailed protein difference, such as deamidation and truncation.
Discover new intact and middle-up approaches for studying protein glycoforms.
Dr. Elena Dominguez Vega Post-Doctoral Researcher, Biomolecular Analysis Group Vrije Universiteit Amsterdam
Dr. Rob Haselberg Post-Doctoral Researcher, Biomolecular Analysis Group, Vrije Universiteit Amsterdam