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Flow injection immunoassay for carcinoembryonic antigen combined with time-resolved fluorometric detection
Authors:Yan Feng  Zhou Jiannong  Lin Jiehua  Ju Huangxian  Hu Xiaoya
Affiliation:College of Chemistry and Chemical Engineering, Yangzhou University, Yangzhou, and Jiangsu Institute of Cancer Prevention and Cure, Nanjing 210009, PR China.
Abstract:Time-resolved fluorescence has been developed for immunoassay to obtain higher sensitivity than usual immunoassays. In this paper, a simple, sensitive and specific method was developed for immunoassay of serum carcinoembryonic antigen (CEA) by combining time-resolved fluoroimmunoassay (TRFIA) and flow injection analysis. Based on a sandwich immunoassay format, a monoclonal antibody immobilized immunoaffinity column inserted in a flow system was used for immunoreactions. The cleaved solution was collected after the reaction between the immunocomplex in the immunoaffinity column and the enhancement solution that was used to cleave the Eu-labels from the immunocomplex, and then detected by time-resolved fluorescence. Serum CEA could be detected in the linear range from 2.5 to 100 ng/ml with a correlation coefficient of 0.997 and the detection limit of 1.0 ng/ml. Twenty human serum samples detected by this method were in good agreement with the results obtained by the electrochemiluminescence immunoassay. This method could be further developed for fast practical clinical detection of serum CEA levels.
Keywords:BSA, Bovine serum albumin   CEA, Carcinoembryonic antigen   CV, Variation coefficient of variation   ECLIA, Electrochemiluminescence immunoassay   EIA, Enzyme immunoassay   FIA, Flow injection analysis   FIIA, Flow injection immunoassay   IRMA, Immunoradiometric assay   PBS, Phosphate buffered solution saline   TM, Tumor marker   TRFIA, Time-resolved fluoroimmunoassay   ECLIA, Electrochemiluminescence immunoassay
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