Chemiluminescence Immunoassay for Quantitative Analysis of Prostate Specific Antigen Complexed toα1-Antichymotrypsin in Human Serum
10.11895/j.issn.0253-3820.150937
- VernacularTitle:化学发光免疫分析法检测血清中前列腺特异抗原复合物
- Author:
Youjun ZHOU
;
Jiaoxia LI
;
Huijun CHENG
;
Qiaofen YANG
;
Meiqiong HE
;
Liping GUO
;
Zhiyong DENG
- Publication Type:Journal Article
- Keywords:
Prostate-specific antigen complexed toα1-antichymotrypsin;
Prostate cancer;
Chemiluminescence enzyme immunoassay;
Horseradish peroxidase
- From:
Chinese Journal of Analytical Chemistry
2016;(8):1209-1214
- CountryChina
- Language:Chinese
-
Abstract:
Eight mouse hybridoma cell lines which stably secreted monoclonal antibodies ( McAbs ) against human prostate-specific antigen-α1-antichymotrypsin complex ( PSA-ACT ) were obtained through hybridoma technique. After purification, the immunological characters of 8 McAbs were identified and classified by epitopes analysis through indirect enzyme-linked immunosorbent assay ( ELISA) . A pair of McAbs was chosen from above 8 McAbs, based on which a highly sensitive, simple and rapid chemiluminescence enzyme immunoassay ( CLEIA) was developed for determination of PSA-ACT in human serums using the lumino-H2 O2 reaction catalyzed by horseradish peroxidase ( HRP) as the chemiluminescence system. Several experiment factors such as coating buffer, coating concentration, dilution ratio of PSA-ACT-HRP complex, incubation time, immunoreaction protocol and chemiluminescence reaction time were optimized. The results showed that the linear range of the proposed method for PSA-ACT determination was 0-40 ng/mL (R2=0. 9943), with the detection limit of 0. 53 ng/mL. The inter-assay relative standard deviations (RSDs) were 4. 6%-6. 6%, and intra-assay RSDs were 5 . 7%-8 . 0%. The recoveries of PSA-ACT at three spiked levels in serum samples were 95. 4%-104. 2%. The proposed method exhibited a cross-reactivity of 0. 6% with free-PSA. The proposed method is stable, sensitive, rapid and simple, and provides a foundation for the development of PSA-ACT CLEIA kit and shows great value in clinical auxiliary diagnosis of prostate cancer.
