| Three cell strains of monoclonal antibodies(MAb) against insulin were obtained. One of the three monoclonal antibodies-INS49Fl, had a high affinity against insulin . INS49F1 and a commercial MAb recoginized different epitopes of insulin , and could be applied in two-site enzyme-linked immunosorbent assay(ELISA).We made a research on the different modes of ELISA for the measurement of insulin. A two-site sandwich ELISA was developed by using two monoclonal antibodies against insulin. One of the antibodies was immobilizd on the microtiter plate, the other was biotinated or labelled with HRP. The sensitivity of the assay was lmlU/L and the recovery was 96. 6%~110. 6%. The intraassay and interassay CVs were 4. 6 % ~ 5. 5 % and 13. 5 % ~ 14. 8 % respectively. The correlation coefficient between measured and expected values was 0.9999 after serial dilutions of the samples with high concentration of insulin. The value for normal samples (n=34) was 0. 9~ 7. 4mIU/L. Compared with RIA,the correlation was y=0. 70X-2. 22, r=0.957, n=90. We also tried to establish a competitive ELISA using PcAb and HRP-labelled second-antibody. |
