亲和吸附纯化甲胎蛋白异质体方法的建立

Establishment of a purificatory method for alpha-fetoprotein variant by affinity adsorption

目的利用小扁豆凝集素（LCA）建立基于微量离心柱的甲胎蛋白异质体L3（AFP—L3）的纯化方法。方法采用硫酸铵分级沉淀的方法由小扁豆中提取凝集素，偶联到活化的琼脂糖凝胶4B（sephrose4B）上，灌注离心柱，制备AFP—L3亲和吸附离心管。血清加入离心管后经吸附和洗脱，便可获得纯化的AFP—L3。随机选取10例AFP阳性患者血清，应用亲和吸附纯化法纯化AFP．L3后应用电化学发光法检测，并与传统的亲和免疫电泳方法相比较。结果10例患者血清经纯化后，AFP-L3检测结果有8例〉5ng／ml，亲和免疫电泳有6例呈现阳性反应。结论成功建立基于微量离心柱的亲和吸附纯化AFP—L3的方法，该方法具有较高的灵敏度，重复性好并且操作简便更利于临床实验室应用。

Objective To establish a purificatory method of alpha-fetoprotein variant （AFP-L3） based on microspincolumn with lens culinaris agglutinin （LCA）. Methods LCA was isolated by ammonium sulfate precipitation method from lens culinaris. AFP-L3 affinity adsorption microspincolumns which were made from LCA coupled with activated Sepharose 4B were prepared. By adding into the centrifuge column, serum was absorbed and eluted to purify AFP-L3. The results of purified AFP-L3 detection of 10 cases AFP positive sera by electro-chemiluminescence immunoassay were compared with traditional crossed affinity immunoelectropboresis. Results 8 of 10 eases AFP-L3 concentration were greater than 5 ng/ml in purified sera. Six cases show positive reaction in affinity immune cross electrophoresis experiment. Conclusion Successfully established purification method of AFP-L3 by affinity absorption based on microspincolumn. The method was more conducive to clinical laboratory applications due to its high sensitive and easy operation.