人组织激肽释放酶活性检测试剂盒的制备  

Construction of an activity assay kit for tissue kallikrein

作　　者：冉晓[1] 游艳艳[2] 张琴[2] 

机构地区：[1]华中科技大学同济医学院附属同济医院急诊科,武汉430030 [2]华中科技大学同济医学院附属同济医院麻醉科,武汉430030

出　　处：《免疫学杂志》2014年第8期737-739,752,共4页Immunological Journal

基　　金：国家自然科学基金(81100864);华中科技大学同济医学院附属同济医院科研基金(2013B004)

摘　　要：目的利用人组织激肽释放酶(TK)能与特异性抗体结合并能催化底物苯甲酰精氨酸乙酯盐酸盐(BAEE)发生化学反应的特性建立1种能检测血浆及尿中TK活性的试剂盒。方法从已有的抗TK的抗体中筛选出1种既能与TK结合又不会封闭其活性位点的抗体作为包被酶标板。然后利用免疫反应及化学显色的方法建立TK活性检测试剂盒。结果抗11P的抗TK的多克隆抗体可作为该试剂盒的包被抗体。成功建立了TK活性检测试剂盒,可用于检测血浆及尿中TK的活性,其标准曲线对线良好(r=0.993)。结论成功建立了1种能检测血浆及尿中TK活性的试剂盒。To prepare a kit for detecting tissue kallikrein（TK） activity in plasma and urine by utilizing immunological characteristics of TK and chemical characteristics of catalyzing benzoyl arginine ethyl ester hydrochloride（BAEE）, we chose an antibody against TK without blocking the active site at first. Then the antibody was pre-coated onto a microplate. After that, a method measuring TK activity by its esterolytic activity over synthetic ester substrates was developed. The result showed that a polyclonal antibody against TK（anti-11P） could be used as capture antibody; The linearity of this kit was demonstrated as 0.993. TK activity in plasma and urine were detected and results showed that TK activity in plasma were apparently higher than that in urine [（2.28±0.61）versus（1.11 ±0.53） Eu/ml, P 0.001]. The assay remained stable, with no change in the values measured, over freezing and thawing. Therefore, the development of a kit for detecting TK activity meets the needs of TK activity detection in urine and plasma.

关 键 词：人组织激肽释放酶 活性 抗体 

分 类 号：R446.6[医药卫生—诊断学]