Citation: ZHOU You-Jun, LI Jiao-Xia, CHENG Hui-Jun, YANG Qiao-Fen, HE Mei-Qiong, GUO Li-Ping, DENG Zhi-Yong. Chemiluminescence Immunoassay for Quantitative Analysis of Prostate Specific Antigen Complexed to α1-Antichymotrypsin in Human Serum[J]. Chinese Journal of Analytical Chemistry, ;2016, 44(8): 1209-1214. doi: 10.11895/j.issn.0253-3820.150937 shu

Chemiluminescence Immunoassay for Quantitative Analysis of Prostate Specific Antigen Complexed to α1-Antichymotrypsin in Human Serum

1.
Nuclear Medicine, Yan'an Hospital Affiliated to Kunming Medical University, Kunming 650051, China;
2.
Nuclear Medicine, The Third Affiliated Hospital of Kunming Medical University, Kunming 650051, China

Received Date: 25 November 2015
Revised Date: 7 June 2016

Figures(2)

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-H₂O₂ 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.
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沈阳化工大学材料科学与工程学院沈阳 110142