压电石英晶体免疫传感器微阵列检测人免疫球蛋白的实验研究

Piezoelectric Immunoglobulin Microarray Immunosensor

目的以AT切型、基频10MHz的镀金石英晶体作为换能器,将抗人免疫球蛋白抗体固定在石英晶体电极表面,用2×5检测池阵列构建一种新型压电石英晶体免疫传感器。方法分别进行标准品及人血清免疫球蛋白的检测,研究了检测温度、时间、特异性等指标对检测的影响。结果该免疫传感器可在15 min内完成检测且无非特异性反应;在检测范围内传感器的响应特性良好,压电晶体频率偏移值与免疫球蛋白浓度呈良好的线性关系,将此免疫传感器用于人血清标本的测定,结果与免疫比浊法符合(r分别为0.94、0.94、0.94、0.92和0.91)。结论此传感器微阵列具有灵敏度高、特异性好,检测通量高,不需标记,操作简便,能实时在线检测和重复使用等优点,可用于临床实验诊断,具有较高的临床推广应用价值。

OBJECTIVE To develop new 2 ×5 type of piezoelectric immunoglobulin microarray immunosensor for determination of immunoglobulin. The energy converters are 10MHz AT-cut quartz crystals with gold-coated electrodes. The monoclonal antibodies of immunoglobulin are immobilized onto the surfaces of crystals gold electrodes by staphylococcal protein A （SPA）. METHODS The standard substance and serum were detected to find the detection time, temperature and specificity of the immunosensor. RESULTS The piezoelectric quartz crystal immunoglobulin microarray immunosensor could complete the detection in 15 rain without nonspecific response. Under the optimized conditions, the experimental results showed that the piezoelectric immunosensor had good response to immunoglobulin whose frequency shifts were linearly dependent on immunoglobulin concentration in different range. The piezoelectric microarray immunosensor had been used to detect immunoglobulin in serum, the analytical results given by this method were in satisfactory agreement with those given by traditional immunoassay, with correlation coefficient of 0. 94, 0. 94, 0. 94, 0. 92 and 0. 91. CONCLUSIONS Piezoelectric microarray immunosensor for the determination of immunoglobulin is of high sensitivity, high specificity, high analysis speed, unnecessary labelling, simple operation, real-time detection, repeated use, etc. It is suitable for detecting of immunoglobulin and should be used for clinical detection.