复合前列腺特异性抗原的单克隆抗体制备及化学发光法免疫定量检测试剂研究  

Generation of monoclonal antibodies against complexed prostate specific antigen and development of an antibody-based chemiluminescence immune quantification assay

作　　者：詹炉停 温桂平[1] 赵敏[1,2] 依含 刘江武 郭小怡 林海军[1,3] 黄刘女 夏宁邵 郑子峥[1,2] 

机构地区：[1]厦门大学分子疫苗学与分子诊断学国家重点实验室,国家传染病诊断试剂与疫苗工程技术研究中心,公共卫生学院,厦门361102 [2]厦门大学生命科学学院,厦门361102 [3]厦门万泰凯瑞生物技术有限公司,厦门361000

出　　处：《中国免疫学杂志》2016年第8期1171-1174,1178,共5页Chinese Journal of Immunology

基　　金：863计划资助项目(No.2011AA02A101;2012AA02A307)

摘　　要：目的：获得特异性检测复合前列腺特异性抗原（c-PSA）的单克隆抗体配对,建立基于化学发光c-PSA免疫定量试剂。方法：利用市购c-PSA抗原免疫6周龄雌性BALB/c小鼠,间接法ELISA差异筛选抗c-PSA、游离前列腺特异性抗原（f-PSA）、总前列腺特异型抗原（t-PSA）的抗体,抗体配对后化学发光定量检测血清中c-PSA蛋白。结果：获得抗体配对1A10/7D6-SAE,经c-PSA标准品及临床血清样本初步筛选,该抗体配对适用于基于化学法发光的免疫反应定量检测试剂的开发;血清阳性样本与阴性样本检测结果显示具有统计学意义（P〈0.000 1）;阳性样本定量结果与Siemens公司复合前列腺特异性抗原测定试剂盒（直接化学发光法）的相关系数为0.97;线性范围为0.1~100 ng/ml;检测灵敏度为0.005 ng/ml。结论：成功筛选获得特异性检测c-PSA的抗体配对,并开发c-PSA化学发光免疫定量试剂,其检测能力与国际主流试剂相当。Objective： To construct a chemiluminescense immune quantification assay based one paired m Abs against complexed prostate specific antigen（ c-PSA）. Methods： Six week-old female BALB / c mice were immunized with the commercial c-PSA antigen. After serum titer reaching a platform stage,the spleen was immunized and fused with mouse myeloma cell lines（ Sp2 /0）. The hybridoma were screened by indirect ELISA,and eight generated antibodies were paired to obtain a quantitative analysis of the chemical luminescence. Results： 7D6 specifically recognized c-PSA,while 1A10 recognized total PSA（ t-PSA）. And the paired antibody 1A10 /7D6 were determined to successfully construct a chemiluminescense immune response quantitative detection method through the detection of c-PSA standard and clinical serum samples. had,positive samples have statistically significant difference（ P〈0. 000 1） with negative samples. And the correlation coefficient R2 was 0. 97 between our c-PSA quantitative results and that of the Siemens c-PSA chemiluminescense immunoassay kit. The detection linear range was 0. 1- 100 ng / ml,and the sensitivity was 0. 005 ng /ml. Conclusion： The paired monoclonal antibodies specifically detecting c-PSA were generated and a c-PSA chemiluminescense immunoassay were developed in this study. The detection capability of our method was comparable with that of the international commercial kit.

关 键 词：复合前列腺特异性抗原 单克隆抗体 化学发光 定量试剂 

分 类 号：R392.33[医药卫生—免疫学]