This application note demonstrates the rapid separation of a wide mass range of peptides and proteins using a reversed-phase monolithic column with both UV and mass spectrometry (MS) detection. The separation is conducted on Thermo Scientific™ ProSwift™ RP-4H columns using conventional water/acetonitrile-based eluents. These simple, straightforward methods can be used to optimize chromatographic separations for analysis time, resolution, and MS detection especially for top-down proteomics.
The Thermo Scientific™ MAbPac™ Protein A column is a high-performance affinity chromatography column specifically designed for the determination of monoclonal antibody concentration in harvest cell culture. The nonporous, hydrophilic stationary phase enables fast and accurate MAb titer analysis. The ability to collect the IgG fraction during the HPLC analysis allows the analyst to characterize the IgG charge variants using a novel pH gradient approach.