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A Novel Chemiluminescence Immunoassay Using Solid‐Phase Antigen for Free 17 β ‐Estradiol in Human Serum
Author(s) -
Qi Yuanyuan,
Chen Hui,
Lin Zhen,
Chen Guonan,
Lin Jinming
Publication year - 2011
Publication title -
chinese journal of chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.28
H-Index - 41
eISSN - 1614-7065
pISSN - 1001-604X
DOI - 10.1002/cjoc.201180424
Subject(s) - chemistry , chemiluminescence , immunoassay , detection limit , chromatography , horseradish peroxidase , bovine serum albumin , analyte , antigen , human serum albumin , conjugate , luminol , chemiluminescent immunoassay , antibody , enzyme , biochemistry , immunology , mathematical analysis , mathematics , biology
A high‐performance chemiluminescence immunoassay, with long‐term durability, good precision and time‐saving, was proposed for the detection of free 17 β ‐estradiol (E 2 ) in human serum. Ninety‐six microplates were coated with bovine serum albumin conjugated E 2 antigen as solid phase for the immunoassay. The E 2 ‐BSA antigen coated on the microplate and the E 2 antigen in the sample competed for the binding sites on the horseradish peroxidase (HRP) labeled anti‐E 2 antibody. Chemiluminescence reaction was subsequently carried out by HRP catalyzing luminol‐H 2 O 2 substrates, and the chemiluminescence intensity was inversely proportional to the amount of analyte in human sera samples. The concentration of immunoreagents, immunoreaction time, and other relevant variable conditions upon the immunoassay were studied and optimized. The proposed method exhibited detection limit as low as 5.94×10 −3 µg·L −1 in a linear detection range from 0.01 to 1.00 µg·L −1 , good recoveries between 105% and 108%, and high precision with intra‐ and inter‐assay coefficients between 7.9% and 14.3%.