Sneekes EJ Webinar Mass spectrometry-based protein characterization can be bracketed into two major applications: top-down proteomics and analysis of recombinant proteins, antibodies, and other biotherapeutics. In each case, the analysis of high molecular weight proteins still can be challenging and the methodology should be chosen to minimize common problems in protein analysis such as degradation, poor recovery, broad elution profiles, poor ionization efficiency, ion sensitivity, and resolution.
We will compare and analyze the performance of a number of different column-spray interfaces to assess their suitability and robustness for intact protein separations. The influence of different reversed-phase chemistry and stationary phase supports (namely, a porous particle, solid core particle and monolithic structure) on separation efficiency is a critical factor. The optimal chromatographic conditions for these column-spray interfaces vary and have to be matched to the mass spectrometric detection. Post-column volume on recovery and sensitivity are analyzed by comparing the data obtained using integrated column-spray devices with traditional setups. Finally, the effects of temperature on the separation and recovery are also discussed.
By the end of the webinar, the participant will be able to perform foolproof intact protein analysis, regardless of their expertise in this area.