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目的:制备两株抗人心肌肌钙蛋白 I(human cardiac troponin I,cTn I)单克隆抗体,建立鲁米诺化学发光酶免疫分析法.方法:制备腹水型单抗、饱和硫酸铵沉淀法和亲和层析柱Protein G纯化抗体,并通过SDS-PAGE和ELISA对抗体特异性进行鉴定,建立鲁米诺化学发光酶免疫分析法,并对30份临床血清样本进行检测.结果:本实验制备的两株单抗(分别命名为Ab1、Ab3;亚型都是IgG1)能够识别以cTn I单体、cTn I-C复合物和cTn I-T-C复合物不同形式存在的cTn I.Ab1、Ab3的效价分别为1:80万和1:40万,亲和力常数分别为1.62×109L/mol、2.60×108L/mol.利用2株单抗建立的鲁米诺化学发光酶免疫分析法检测范围为:6.25 ~400 ng/ml.对30份临床样本进行检测,阳性检出率为77.3%,阴性检出率为100%.结论:建立了可以检测以cTn I单体、cTn I-C和cTn I-T-C复合物形式存在的cTn I的鲁米诺化学发光酶免疫分析法,具有更高的实用性.“,”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.