抗人白细胞间素—2McAb亲和力排列及亲和指数的测定

The Determination of Affinity Indexes and Affinity Ranking of Monoclonal Antibodies against Interleukin-2

利用蛋白浓度校正的ELISA稀释曲线快速确定了抗人白细胞间素-2(IL-2)单克隆抗体(McAb)的亲和力排列。该排列与用竞争ELISA测定的亲和常数(K_D值)相一致。为了更实际地确定McAb的用途,又用十二烷基磺酸钠(SDS)等解离因素,建立了反映 McAb在实际应用时亲和力变化强度的吸附-解离ELISA,测定McAb的亲和指数(ID)。根据亲和力排列和亲和指数可更准确地取舍McAb,供检测或亲和色谱使用。

The affinities of the monoclonal antibodies (McAb) against human interleu-kin-2 were ranked easily by their protein concentration-adjusted ELISA dilution curves. The ranking was consistent with the'ranking of their KD values measured by a competitive ELISA method. In order to reflect the virtual affinity changes of the McAb in practical uses, an absorption elution sandwich ELISA assay was established to obtain the affinity indexes, which were the ratio of absorbance value produced by one McAb in normal antigen-absorbing condition to that in elution condition ( with low concentration of detergents or low pH ) . By affinity ranking and affinity indexes, we can choose more proper antibodies used either for detection assay or for affinity chromatography.