ABSTRACT
Objective: To prepare monoclonal antibodies (McAb) with cardiac troponin I (cTnI) which was purified from fresh human cardiac muscle within 6 h. Methods: (1) Extraction and purification of human cTnI: cTnI was purified by high salt extraction, saltless precipitation, 65℃ treatment, ammonium sulfate fractionation and DEAE-cellulose chromatography, etc. (2) Preparation of anti human cTnI McAb: The purified cTnI was injected into the spleen of BALB/c mice. The cTnI-primed spleen cells were fused with Sp2/0 myoloma cell. The McAbs anti human cTnI were obtained by screening with indirect ELISA and 3 times clone. (3)The identification of anti cTnI McAb. Results: Five hybridoma cell lines, named 3A7，3A11，3D2，3F10 and 1H9 were developed, which could secret McAb stably. The 5 McAbs all were demonstrated to be IgG2a by double gel diffusion test. The number of hybridoma chromosomes was between 92 to 110 and the chromosomes were mainly telocentric. Five kinds of ascites had no cross-reaction to LDH，CK，CK-MB ，AST and cardiac troponin T(cTnT), and their titers were between 3.2×10-6 to 1.6×10-7. Conclusion: 3D2，3F10 and 3A7,3A11,1H9 react to different epitopes of cTnI.
ABSTRACT
Objective:To develop a sensitive,rapid,simple gold immunochromatography assay for human cardiac troponin I(cTnI).Methods:The cTnI was used to immunize Balb/c mice by route method.The spenocytes were fused with SP2/0 myeloma cell lines.Six hybridoma cell lines,secreting stably McAbs to cTnI antigen have been obtained and screened.The 3F10 was coupled with colloidal golds,the 3A7 biotinylated.Streptavidin was coupled with the nitrocellose strip to prepare an test apparatu.According to red line in nitrocellose,thus can analysis the results.Results:The sensitivity of cTnI measured by this apparatus was 0.4 ng/ml.The 30 patients with acute myocardial infarction(AMI)were detected for cTnI by this apparatus and by the product of PBM Co USA as standard.The coincidence rate of both two assays was 93.6%. Conclusion:This method is a sensitive,specific simple and rapid assay.