ABSTRACT
Objective:Two anti-human cardiac troponin I(cTnI)monoclonal antibodies were prepared to establish Luminol chemiluminescence enzyme immunoassay.Methods: Obtaining ascites monoclonal antibody was purified by ammonium sulfate fractionation and protein G affinity chromatography column, and their specificity was identified by SDS-PAGE and ELISA methods.Luminol chemiluminescence enzyme immunoassay was successfully established,which was used to detect 30 clinical serum samples.Results:Two antibodies named Ab1 and Ab3 could recognize different forms of cTnI including cTn I monomers,cTn I-C complexes and cTn I-T-C complexes.Both Ab1 and Ab3 are IgG1.The titers of the Ab1 and the Ab3 were up to 1:800 000 and 1:400 000,respectively.The affinity constants of Ab1 and Ab3 were 1.62×109L/mol and 2.60×108L/mol,respectively.The detection range of Lumino chemiluminescence enzyme immunoassay established by two monoclonal antibodies was 6.25 ng/ml to 400 ng/ml. Detecting 30 clinical samples attained the positive detection rate of 77.3% and the negative detection rate of 100%.Conclusion:The chemilu minescence enzyme immunoassay has been established to detect different forms of cTn I including cTn I monomer,cTn I-C complexes and cTn I-T-C complexes,which has better practicality.