MLCK定量检测的双抗体夹心ELISA方法建立  

Estabilishment of Double-antibody Sandwich Enzyme Linked Immunosorbent Assay for Quantitative Detection of Mylosin-light-chain Kinase

作　　者：邸亚男[1,2] 王晋星一[3] 王猛[3] 李兴[1] 杨国珍[1] 潘卫[1,2] 

机构地区：[1]贵阳医学院医学检验系,贵州贵阳550004 [2]贵阳医学院附院临床生化科,贵州贵阳550004 [3]贵阳医学院基础医学(生物技术方向),贵州贵阳550004

出　　处：《贵阳医学院学报》2011年第3期289-291,共3页Journal of Guiyang Medical College

基　　金：贵州省科学技术基金资助项目[黔科合J字(2011)2118号];贵州省教育厅自然科学研究项目;黔教科[2010]032贵州省教育厅;黔教高发[2006]354号

摘　　要：目的:建立定量检测人肌球蛋白轻链激酶(MLCK)的双抗夹心ELISA法。方法:用抗MLCK兔多克隆抗体包被酶标板,1%小牛血清白蛋白(1%BSA)作为封闭液,抗MLCK羊多克隆抗体和HRP-兔抗羊抗体分别为包被抗体和检测抗体,建立双抗体夹心ELISA法,并进行了实验条件的优化;同时对新建立方法的灵敏度、精密度进行了评价。结果:包被抗体和检测抗体最佳工作浓度分别为1∶2 000和1∶200,该方法的敏感性为5.16 mg/L,低、高浓度样本批内和批间变异系数分别为5.9%、6.1%和16.0%、19.0%。结论:建立了MLCK定量检测的ELISA方法,其灵敏度和精密度均较好,为人血清MLCK的检测提供了一种简便快捷方法。Objective：To establish a double-antibody sandwich enzyme linked immunosorbent assay（ELISA） method for quantitative detection of mylosin-light-chain kinase（MLCK） in human serum.Methods：Double-antibody sandwich ELISA was carried out by using rabbit anti-MLCK polyclonal antibody coating assay plates,1% BSA as blocking solution,goat anti-MLCK polyclonal antibody and HRP rabbit anti-goat antibody as coating antibody and detecting antibody respectively.Experiment condition was optimized.Sensibility and precision of the assay were evaluated.Results：The optimal working concentrations of capture antibody and detecting antibody were 1∶2000 and 1∶200 respectively.Sensitivity of the assay was 5.16 ng/ml;Variation coefficients of low and high concentration samples between different batches and in the same batch were 5.9%,6.1% and 16.0%,19.0% respectively.Conclusions：ELISA method is successfully established for quantitative detection of MLCK with high sensibility and precision,and it provides an easy and efficient method for MLCK detection in human serum.

关 键 词：肌球蛋白轻链激酶 酶联免疫吸附测定 生物技术 

分 类 号：R34-33[医药卫生—基础医学] R345