双抗体C-ELISA测定人血清胰岛素的实验研究

An Experimental Study on Measurement of Human Serum Insulin with Two Antibody C-ELISA

我们建立了双抗体竞争抑制法 ELISA(C-ELISA)测定人血清胰岛素的方法。第1抗体和第2抗体的最佳工作浓度分别为1:5和1:1280;胰岛素浓度经过平方根对数变换后与吸光度呈良好的直线关系(r=-0.99,P=0.01);线性范围为1～640mU/L;灵敏度为1mU/L;批内和批间变异分别为9.5%和12.5%;与 RIA 法相关良好。C-ELISA,测定人血清胰岛素勿需特殊设备,无放射性污染,操作简便,灵敏度高,可成为 RIA 法的理想替代方法。

A new method was estabolished for measurement of human serum insulin concentration, which base on the principle of competitive satuation ELISA(C-ELISA),and charactized by in- dirct binding of an anti-insulin antibody by an antiglobin antibody.The optimal dilution of anti insulin antibody and antigolbin antibody is 1:5,1:1 280,respectively.The logarithm of the square root of serum insulin concentration is corelative with the optical absorbance(r=-0.99, P=0.01).This new method have a wild standard curve ranging from 1mU/L to 640 mU/L in- sulin.It's sensitivity is 1mU/L insulin,the between-and within-assay coefficients of variation is respectively less 12.5% and 9.5%.The results from C-ELISA is highly eorelated to from RIA,so C-ELISA may come to be a good substitute of RIA.