Capillary electrophoretic immunoassay for alpha-fetoprotein with chemiluminescence detection.
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.
Liu
Yan-Ming
YM
College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang, China. liuym9518@sina.com
Mu
Hai-Bei
HB
Zheng
Yan-Li
YL
Wang
Cheng-Quan
CQ
Chen
Yong-Hong
YH
Li
Fu-Rong
FR
Wang
Jun-Hua
JH
Cheng
Jie-Ke
JK
D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008163:Luminescent Measurements / Q000379:methods*; D000509:alpha-Fetoproteins / Q000032:analysis* / Q000302:isolation & purification
D000509:alpha-Fetoproteins