A New Method to Control the Coupling Site of Protein Ligands and Affinity Chromatographic Media
亲和层析介质上蛋白质偶联位点的控制方法

Lysozyme was coupled on Sepharose 4FF media activated with 1,4-butanediol diglycidyl ether. HPLC-MS and enzymatic digestion were used to identify the coupling sites of lysozyme on the media. The effects of epoxy group density and the coupling reaction condition on the coupling sites were investigated. It was found that K96 was the main coupling site on the media when the epoxy group density was 11.34μmol/g and pH 9.5. The coupling reaction time did not affect the coupling site. Epoxy group density and pH value during the coupling reaction are the main factors affecting the coupling sites of lysozyme on the media. More lysozyme molecules were coupled on the media via K33 and K97 when pH value was above 10.5.