摘要
A high-performance chemiluminescence immunoassay, with long-term durability, good precision and time- saving, was proposed for the detection of free 17β-estradiol （E2） in human serum. Ninety-six microplates were coated with bovine serum albumin conjugated E2 antigen as solid phase for the immunoassay. The E2-BSA antigen coated on the microplate and the E2 antigen in the sample competed for the binding sites on the horseradish peroxi- dase （HRP） labeled anti-E2 antibody. Chemiluminescence reaction was subsequently carried out by HRP catalyzing luminol-H2O2 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 1.00μg·L^-1 good recoveries between 105% and 108%, and high precision with intra- and inter-assay coefficients between 7.9% and 14.3%.
A high-performance chemiluminescence immunoassay, with long-term durability, good precision and time- saving, was proposed for the detection of free 17β-estradiol （E2） in human serum. Ninety-six microplates were coated with bovine serum albumin conjugated E2 antigen as solid phase for the immunoassay. The E2-BSA antigen coated on the microplate and the E2 antigen in the sample competed for the binding sites on the horseradish peroxi- dase （HRP） labeled anti-E2 antibody. Chemiluminescence reaction was subsequently carried out by HRP catalyzing luminol-H2O2 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 1.00μg·L^-1 good recoveries between 105% and 108%, and high precision with intra- and inter-assay coefficients between 7.9% and 14.3%.
基金
Project supported by the National Natural Science Foundation of China （Nos. 90813013, 2U933UU2）.
