Capillary electrophoretic immunoassay for alpha-fetoprotein with chemiluminescence detection |
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Authors: | Liu Yan-Ming Mu Hai-Bei Zheng Yan-Li Wang Cheng-Quan Chen Yong-Hong Li Fu-Rong Wang Jun-Hua Cheng Jie-Ke |
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Institution: | College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang, China. liuym9518@sina.com |
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Abstract: | A capillary electrophoretic immunoassay with chemiluminescence detection (CEIA-CL) using a non-competitive format for analyzing tumor marker alpha-fetoprotein (AFP) has been developed. In this method, antigen (Ag) AFP reacts with an excess amount of horseradish peroxidase (HRP)-labeled antibody (Ab*). The free Ab* and the bound Ab*-Ag complex produced in the solution are separated by CE in a separation capillary. Then they catalyze the reaction of enzyme substrate luminol and H(2)O(2) in a reaction capillary following the separation capillary. Parameters affecting the CE separation and CL detection were investigated. Under the optimal conditions, the free Ab* and the Ab*-Ag complex were well separated within 4 min, the linear range and the detection limit (S/N=3) for AFP were 5-500 ng/ml and 0.85 ng/ml (1.2 x 10(-11)M), respectively. The proposed method has been applied satisfactorily in the analysis of human sera samples. |
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