CAJ | 학술논문

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(sephrose 4B)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＞5 ng/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.
목적 이용소편두응집소(LCA)건립기우미량리심주적갑태단백이질체L3(AFP-L3)적순화방법.방법 채용류산안분급침정적방법유소편두중제취응집소,우련도활화적경지당응효4B(sephrose 4B)상,관주리심주,제비AFP-L3친화흡부리심관.혈청가입리심관후경흡부화세탈,편가획득순화적AFP-L3.수궤선취10례AFP양성환자혈청,응용친화흡부순화법순화AFP-L3후응용전화학발광법검측,병여전통적친화면역전영방법상비교.결과 10례환자혈청경순화후,AFP-L3검측결과유8례＞5 ng/ml,친화면역전영유6례정현양성반응.결론 성공건립기우미량리심주적친화흡부순화AFP-L3적방법,해방법구유교고적령민도,중복성호병차조작간편경리우림상실험실응용.
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 immunoelectrophoresis.Results 8 of 10 cases 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.