Generation of monoclonal antibodies against complexed prostate specific antigen and development of an antibody-based chemiluminescence immune quantifica-tion assay
10.3969/j.issn.1000-484X.2016.08.019
- VernacularTitle: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
- Author:
Luting ZHAN
;
Guiping WEN
;
Min ZHAO
;
Han YI
;
Jiangwu LIU
;
Xiaoyi GUO
;
Haijun LIN
;
Liunü HUANG
;
Zizheng ZHENG
- Publication Type:Journal Article
- Keywords:
Complexed prostate specific antigen ( c-PSA);
Monoclonal antibody;
Chemiluminescense;
Quantification reagent
- From:
Chinese Journal of Immunology
2016;32(8):1171-1174,1178
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct a chemiluminescense immune quantification assay based one paired mAbs 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 R 2 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 .
