Preparation of a novel dual-function strong cation exchange/hydrophobic interaction chromatography stationary phase for protein separation

► We explored a bifunctional column with ligand containing sulfonic and benzyl groups. ► This column can provide two operation modes (HIC and SCX). ► High resolution and selectivity are obtained in both SCX and HIC modes, respectively. ► This column can replace two corresponding single mode (SCX and HIC mode) columns. ► Based on this bifunctional column, 2DLC was established using only a single column.