Document Detail

Development of a high-throughput, indirect antibody immobilization format chemiluminescence enzyme immunoassay (CLEIA) for the determination of progesterone in human serum.
MedLine Citation:
PMID:  18452138     Owner:  NLM     Status:  MEDLINE    
A high-throughput and simple chemiluminescence (CL) enzyme immunoassay (CLEIA) for the determination of progesterone (P) in human serum was developed, with the highly sensitive 4-methoxy-4-(3-phosphatephenyl)-spiro-(1,2-dioxetane-3,2'-adamantane) (AMPPD)-alkaline phosphatase (ALP) system as the CL detection system. The results showed that the indirect immobilization of rabbit anti-progesterone polyclonal antibody (RAPA) through secondary antibody exhibited apparent advantages over direct coating in terms of antibody saving and improvement of the coating stability and uniformity. The direct analysis of P in human serum without extraction was realized by using 8-anilino-1-naphthalenesulphonic acid (ANS) to displace P from its binding proteins. The effect of several relevant parameters of the immunoreaction were examined and optimized. Compared with some commercial progesterone kits, the presented CLEIA has higher sensitivity with detection limitation as low as 0.06 ng/mL. The recoveries were 95.9-101%. The coefficient of variation was <8.4% and 9.9% for intra- and inter-assay precision, respectively. This method has been successfully applied to the evaluation of P in human serum.
Shiqi Ren; Xu Wang; Zhen Lin; Zhenjia Li; Xitang Ying; Guonan Chen; Jin-Ming Lin
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Luminescence : the journal of biological and chemical luminescence     Volume:  23     ISSN:  1522-7243     ISO Abbreviation:  Luminescence     Publication Date:    2008 May-Jun
Date Detail:
Created Date:  2008-05-22     Completed Date:  2008-07-16     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100889025     Medline TA:  Luminescence     Country:  England    
Other Details:
Languages:  eng     Pagination:  175-81     Citation Subset:  IM    
Copyright Information:
(c) 2008 John Wiley & Sons, Ltd.
Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology, Ministry of Education, Department of Chemistry, Tsinghua University, Beijing 100084, People's Republic of China.
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MeSH Terms
Alkaline Phosphatase
Chemiluminescent Measurements
Immunoenzyme Techniques / methods*
Progesterone / blood*
Reg. No./Substance:
0/Antibodies; 57-83-0/Progesterone; EC Phosphatase

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