基于蛋白A定向固定抗体的触珠蛋白压电免疫传感器的研究

Piezoelectric Immunosensor for the Detection of Haptoglobin Using the Protein A-Based Oriented Immobilization of Antibody

用蛋白A-金电极法将触珠蛋白抗体固定在9 MHz镀金石英晶体比表面,研制了一种简单、灵敏的压电免疫传感器,用于溶血性贫血与严重肝病标志物-触珠蛋白的高特异性检测.比较了传感器采用直接金电极吸附法、牛血清白蛋白吸附-戊二醛交联法和蛋白A法三种固定抗体的方法.研究了蛋白A的定向自组装、抗体的固定化浓度和免疫反应时间等主要因素对免疫传感检测灵敏度的影响,并考察了传感器响应与再生性能.测得该免疫传感器检测人血清中触珠蛋白的线性浓度范围为1.27～31.7 mg/L.

A piezoelectric immunosensor has been developed for the detection of Haptoglobin（Hp） by immobilization at anti-Hp antibody adopting protein A method to the 9 MHz quartz At-cut with gold coated electrodes. Three methods for immobilization of anti-Hp antibody on the gold electrodes were employed. The protein A method gave the best results in terms of sensitivity, stability and reproducibility. The factors affecting the sensitivity of the system such as concentration of protein A coated, the titer of antibody, the reaction time of immunoreaction etc., were discussed. The linear range for Haptoglobin was 1.27 - 31.7 mg/L.