化学发光免疫分析法检测血清中前列腺特异抗原复合物

发布时间：2018-07-03 09:09

本文选题：前列腺特异抗原 + 前列腺癌　；参考：《分析化学》2016年08期

【摘要】：采用杂交瘤技术构建抗前列腺特异抗原-α1-抗胰凝乳蛋白酶(Prostate specific antigen-α1-antichymotrypsin complex,PSA-ACT)单克隆抗体(单抗)细胞株,共获得8株稳定分泌PSA-ACT单抗的细胞株,纯化后进行免疫特性鉴定并做单抗配对。选择其中一对单抗,采用辣根过氧化物酶(HRP)催化鲁米诺(lumino)-H_2O_2化学发光体系,建立了测定人血清中PSA-ACT浓度的化学发光酶免疫分析方法(CLEIA)。对包被缓冲液、包被抗体浓度、酶标抗体稀释度、温育时间、发光反应时间和免疫反应步骤等实验参数进行了优化。在最佳实验条件下,PSA-ACT浓度在5~40 ng/m L范围内线性关系良好(R~2=0.9943),检出限为0.53 ng/m L,批内相对标准偏差(RSD)为4.6%~6.6%,批间RSD为5.7%~8.0%,回收率为95.4%~104.2%,与游离前列腺特异性抗原F-PSA的交叉反应率为0.6%。本方法简单、稳定、灵敏、快速,为开发检测PSA-ACT的CLEIA试剂盒奠定了基础。
[Abstract]:Hybridoma technique was used to construct prostate specific antigen- 伪 1-antichymotrypsin complex PSA-ACT monoclonal antibody cell lines. Eight cell lines secreting stable PSA-ACT monoclonal antibodies were obtained. A chemiluminescent enzyme immunoassay (CLEIA) for the determination of PSA-ACT in human serum was developed by using a pair of monoclonal antibodies and a chemiluminescence system of (lumino) / S _ 2O _ 2 catalyzed by horseradish peroxidase (HRP). The experimental parameters such as coating buffer concentration of coated antibody dilution of enzyme-labeled antibody incubation time luminescence reaction time and immunoreaction steps were optimized. Under the optimum experimental conditions, the concentration of PSA-ACT was linear in the range of 5o 40 ng/m / L (RPSA-ACT 0.9943), the detection limit was 0.53 ng/m / L, the relative standard deviation (RSD) was 4.6 ng/m / L, and the relative standard deviation (RSD) was 4.66.0.The recovery rate was 95.4104.22.The cross reaction rate with free prostate specific antigen F-PSA was 0.6m. The method is simple, stable, sensitive and rapid, which lays a foundation for the development of CLEIA kit for the detection of PSA-ACT.
【作者单位】：昆明医科大学附属延安医院核医学科;昆明医科大学第三附属医院核医学科;
【基金】：云南省应用基础研究(昆医联合专项)项目(No.2014FB077)资助
【分类号】：R446.6

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