“Click chemistry” preparation of WCX packings for protein separation

Click chemistry was applied to immobilize three kinds of alkyne-carboxylic acids onto azide-modified silica gel to prepare three novel stationary phases for weak cation exchange chromatography (WCX). The developed protocol combines the benefits of operational simplicity, exceptionally mild conditions and high surface loadings. Six kinds of standard proteins were separated completely on the novel packings. Compared with commercial WCX columns, the three kinds of novel WCX packings prepared by click chemistry approach have better resolution and selectivity. Lysozyme was purified successfully from egg white with the novel WCX column by one step. The purity was more than 97% and a high specific activity was achieved to be 81,435 U/mg. The results illustrate the potential of click chemistry for preparation of stationary phase for IEC.