人血清真胰岛素酶联免疫法的建立及初步应用

DEVELOPMENT AND PRELIMINARY APPLICATION OF ENZYMELINKED IMMUNOSORBENT ASSAY FOR HUMAN NET INSULIN IN SERUM

人血清真胰岛素酶联免疫法的建立及初步应用黎明吴从愿宋爱玲张葵摘要选用两种可相互配对的抗人胰岛素（ｈＩｎｓ）单克隆抗体，其中一种包被塑料孔条作固相抗体，另一种经生物素化后作液相抗体，再以辣根过氧化物酶标记的亲和素放大检测信号，建立了高度特异和灵敏的人血...

A specific and sensitive monoclonal antibody_based two_site enzyme immunoassay (BA_ELISA) was described for human net insulin in serum. Two monoclonal antibodies (McA) against insulin were used; one was absorbed to polystyrene microtitre strip wells as solid phase and the other was biotinated, thus quadravalencely binding to avidin_horseradish peroxidase to amplify the detecting signal during the one step BA_ELISA. With a detection limit of 0.8mU/L, the assay covered a working range of 1.5～160.0mU/L, and did not cross_react with high dosage of human proinsulin (2000pmol/L) and C peptide (5000pmol/L). The mean recovery was 101.7% and the intraassay cv and inter_assay cv were 4.1% and 7.0%, respectively. Serum insulin levels both in normal and type 2 diabetes mellits patients, as detected by the present method, showed a good correlation with those measured by RIA, r being 0.914 and 0.964 respectively. These results suggest that the BA_ELISA has more advantages than the routine RIA in estimating hIns.