| 高特异性人血清白蛋白化学发光免疫分析方法的建立; Establishment of a High-specificity Chemiluminescence Immunoassay Assay for Human Serum Albumin | |
| 张雅丽 ; 袁伦志 ; 王腾云 ; 刘旋 ; 吴坤 ; 张军 ; 程通 ; 夏宁邵 | |
| 2014-05-28 | |
| 关键词 | 人血清白蛋白(HSA) 单克隆抗体 种属特异性 化学发光免疫分析(CLIA) human serum albumin(HSA) monoclonal antibody species-specific chemiluminescence immunoassay(CLIA) |
| 英文摘要 | 人血清白蛋白(HSA)是人类肝脏功能的重要生物标志物.本研究旨在发展一种具有高度种属特异性和灵敏度的HSA化学发光免疫分析(HSA-ClIA)定量检测方法.通过HSA免疫小鼠,制备了6株HSA鼠单克隆抗体;使用间接化学发光检测(IndIrECT-ClIA)和WEST blOTTIng检测方法评价单抗对HSA的反应活性;使用免疫组织化学检测(IHC)和WEST blOTTIng检测方法评价单抗的种属特异性;选择最佳的包被抗体和检测抗体配对;确定HSA-ClIA的定量线性和范围.WEST blOTIng和IHC方法中,4f5和5d2抗体反应性最强,且不与小鼠、大鼠、牛、兔、山羊、猴、鸡等常见实验动物血清白蛋白交叉反应.以单抗5d2配合单抗4f5建立的HSA-ClIA,线性范围达到122~31 250Pg/Ml,可适用于多种样本的HSA定量检测.本研究发展的单克隆抗体和检测方法为HSA的特异性检测提供了重要工具和手段.; Human serum albumin(HSA)is an important bio-marker for liver function.This study aimed to develop aquantitative species-specific chemiluminescence immunoassay(CLIA)with high sensitivity for detection of HSA.A total of 6 strains of monoclonal antibodies(mAbs)against HSA were obtained by mouse immunization.Subsequently,the reactivity of mAbs against HSA was evaluated by indirect CLIA and western blotting,and the species-specificity of mAbs was performed by western blotting and immunohistochemistry(IHC).And then,the best capture antibody and the detection antibody pairs were selected.Finally,the quantitative linearity and range of HSA-CLIA had been determined.Among these antibodies,the mAb 4F5 and 5D2 had the strongest reactivity against HSA,and did not present cross-reactivity with serum albumin from common experimental animals,such as mouse,rat,bovine,rabbit,goat,monkey,and chicken.Based on the mAb pair of 5D2 and 4F5,we developed an HSA chemiluminescence immunoassay with a linear range from 122 pg/mL to 31 250pg/mL,which was suitable for HSA quantification of various specimens.The monoclonal antibodies and the detection methods we developed in this report would provide important tools for the specific detection of HSA.; 国家重大科技专项(2012ZX10004503-005) |
| 语种 | zh_CN |
| 内容类型 | 期刊论文 |
| 源URL | [http://dspace.xmu.edu.cn/handle/2288/119240]  |
| 专题 | 生命科学－已发表论文 |
| 推荐引用方式 GB/T 7714 | 张雅丽,袁伦志,王腾云,等. 高特异性人血清白蛋白化学发光免疫分析方法的建立, Establishment of a High-specificity Chemiluminescence Immunoassay Assay for Human Serum Albumin[J],2014. |
| APA | 张雅丽.,袁伦志.,王腾云.,刘旋.,吴坤.,...&夏宁邵.(2014).高特异性人血清白蛋白化学发光免疫分析方法的建立.. |
| MLA | 张雅丽,et al."高特异性人血清白蛋白化学发光免疫分析方法的建立".(2014). |
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