贵州医科大学学报

2011, v.36;No.150(03) 289-291

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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

邸亚男;王晋星一;王猛;李兴;杨国珍;潘卫;
DI Yanan1,2,WANGJIN Xingyi3,WANG Meng3,LI Xing1,YANG Guozhen1 PAN Wei1,2 (1.Department of Clinical Laboratory Science,Guiyang Medical College,Guiyang 550004,Guizhou,China; 2.Department of Clinical Biochemistry Laboratory,the Affiliated Hospital of Guiyang Medical College, Guiyang 550004,Guizhou,China;3.Department of Biological Technology,Guiyang Medical College,Guiyang 550004,Guizhou,China.)

摘要(Abstract):

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(KeyWords): 肌球蛋白轻链激酶;酶联免疫吸附测定;生物技术
mylosin-light-chain kinase;enzyme-linked immunosorbent assay;biotechnology

Abstract:

Keywords:

基金项目(Foundation): 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

作者(Author): 邸亚男;王晋星一;王猛;李兴;杨国珍;潘卫;
DI Yanan1,2,WANGJIN Xingyi3,WANG Meng3,LI Xing1,YANG Guozhen1 PAN Wei1,2 (1.Department of Clinical Laboratory Science,Guiyang Medical College,Guiyang 550004,Guizhou,China; 2.Department of Clinical Biochemistry Laboratory,the Affiliated Hospital of Guiyang Medical College, Guiyang 550004,Guizhou,China;3.Department of Biological Technology,Guiyang Medical College,Guiyang 550004,Guizhou,China.)

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DOI: 10.19367/j.cnki.1000-2707.2011.03.020

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