一种定性比较抗体相对亲和力的酶联免疫吸附测定方法

目的:建立一种简便的对抗体相对亲和力进行定性比较的酶联免疫吸附测定(ELISA)方法,以便快速、简便地从大量抗体突变体中挑选高亲和力突变体。方法:将待测抗体倍比稀释后用直接ELISA方法进行定量,同时用相同浓度抗体作为一抗与抗原进行间接ELISA反应,以前者吸光度值为横轴、后者吸光度值为纵轴绘制散点图,通过拟和后的曲线判断抗体亲和力高低,并通过BIA-core法对该方法的准确性进行验证。结果:通过该方法获得的抗体亲和力高低情况与经测定抗体亲和力得出的结果一致。结论:该ELISA方法是一种简便可行、准确有效的抗体亲和力定性比较方法,可以应用于不同抗体的亲和力成熟比较研究。

A Convenient Enzyme Linked Immunosorbent Assay Method for Antibody Affinity Qualitative Comparison

Objective：To approach an effective enzyme linked immunosorbent assay（ELISA） method for screening high-affinity mutants from the amount of variants conveniently.Methods：Serial delusions of antibodies were used as antigen in the direct ELISA and the absorbance of direct ELISA represented concentration of the diluted sam-ples.Indirect ELISA was carried out with the diluted samples simultaneously.Then relative lines were drawn by taking the direct ELISA absorbance as X axle and the Indirect ELISA absorbance as Y axle to analysis the affini-ty of antibody samples.The results were compared with the data from BIA-core.Results：The results of this method were consistent with that of BIA-core which is more precise.Conclusion：The method is a convenient means to compare affinity of antibody mutants qualitatively,and it can be used in the process of antibody affinity maturation in vitro.