Using Core-Shell UHPLC Columns for Improved Separation and Characterisation of Immunoglobulins and Other Large Intact Proteins

The widespread popularity of developing monoclonal antibodies and other similar immunoglobulin-like proteins as human therapeutics has required new tools for reversed phase LC/MS analysis of proteins. Existing fully porous column technologies have struggled in resolving chemical and structural details of such large, hydrophobic proteins. Recently introduced UHPLC column technologies including wide-pore core-shell particle columns have addressed such issues by demonstrating increased peak capacity as well as improved recovery of hydrophobic proteins. Several examples of the utility of such materials are shown and important method development considerations are discussed which address the radically different chromatographic characteristics of such materials.