纯化小鼠腹水中抗富含组氨酸糖蛋白单克隆抗体方法的比较

THE COMPARISON OF THREE METHODS FOR PURIFICATION OF MONOCLONAL ANTIBODY AGAINST HISTIDINE-RICH GLYCOPROTEIN FROM MOUSE ASCITES

用羟基磷灰石柱层析法，辛醚－硫酸铵沉淀法以及辛酸－硫酸按沉淀法加羟基磷灰石柱层析法从小鼠腹水中提取纯化抗富含组氨酸糖蛋白单克隆抗体。结果表明，羟基磷灰石柱层析法简便，快速，其单克隆抗体产量和纯度均高，适合大多数实验室采用。

sing hydroxylapatite column chromatography(HACC),caprylic acid-ammonium sulphateprecipitation(CAASP),and caprylic acid-ammonium sulphate precipitation plus hydroxylapatitecolumn chromatography(CAASP+HACC),monoclonal antibody(MoAb)against histidine-richglycoprotein from mouse ascites was purified.The MoAb obtained from 1 ml of mouse ascites byNACC,CAASP,and CAASP+HACC amounted to 16.60,7.95,3nd 7.20mg respectively.Only two electrophoretic. bands (heavy and light chains of IgG)were seen on SDS-PAGE profileof MoAb purified with HACC or CAASP+HACC;whereas there were at least three bands onSDS-PAGE profile of MoAb purified with CAASP. The results indicated that HACC was simpleand rapid in procedures,with high yields and purity of MoAb.Therefore,it is suggested thatHACC was suitable to most laboratories.