Acta Chimica Sinica ›› 2006, Vol. 64 ›› Issue (4): 325-330. Previous Articles     Next Articles

Original Articles

基于多层酶/纳米金固定甲胎蛋白免疫传感器的研究

黎雪莲,袁若*,柴雅琴,朱强,张凌燕,王娜   

  1. (西南大学化学化工学院 重庆市现代分析化学重点实验室 重庆 400715)
  • 收稿日期:2005-07-08 修回日期:2005-10-16 出版日期:2006-02-28 发布日期:2006-02-28
  • 通讯作者: 袁若

Reagentless Amperometric Immunosensor for α-1-Fetoprotein Based on Multilayers of Horseradish Peroxidase/Nano Au

LI Xue-Lian, YUAN Ruo*, CHAI Ya-Qin, ZHU Qiang, ZHANG Ling-Yan, WANG Na   

  1. (Chongqing Key Laboratory of Analytical Chemistry, College of Chemistry and Chemical Engineering,
    Southwest University, Chongqing 400715)
  • Received:2005-07-08 Revised:2005-10-16 Online:2006-02-28 Published:2006-02-28
  • Contact: YUAN Ruo

An immunosensor for direct amperometric determination of α-1-fetoprotein (AFP) was constructed by α-1-fetoprotein antibody (anti-AFP) immobilized on multilayers of horseradish peroxidase (HRP)/nano Au through electrostatic adsorption onto L-cysteine modified gold electrode surface. The fabricated procedures and electrochemical behaviors of proteins on such an interface were characterized with electrochemical impedance spectroscopy, cyclic voltammetry and chronoamperometry. The determination of AFP was established by chronoamperometry to record the direct electrochemistry response of HRP to reduction of H2O2. Under optimal conditions, the immunosensor was highly sensitive to AFP with a detection limit of 0.5 ng•mL-1 and the linear range was at two concentrations from 1 to 10 and from 10 to 200 ng• mL-1 with 0.7 mmol•L-1 H2O2 in analyte solution. The dependence of the number of HRP/nano Au layer, pH, incubation time, reproducibility and stability was studied. More than 86.7% of the results of the human serum samples obtained by this method were in agreement with those obtained by radioimmunoassay.

Key words: reagentless immunosensor, α-1-fetoprotein, horseradish peroxidase, nano Au, electrostatic adsorption