新型压电石英晶体免疫传感器抗体固定方法的建立

Establishment of a Novel Antibody Immobilizati on Method on Piezoelectric Quarts Crystal Immunosensor

目的 探讨并建立一种高效免疫微阵列传感器抗体固定的方法。方法 分别采用普通蛋白A(SPA)法与经巯基取代SPA(自组装SPA)法固定石英晶体微阵列上抗体,并用石英晶体微天平(QCM)分别检测其不同浓度抗体、抗原条件下发生抗原抗体反应,并对其抗体包被量、反应频率漂移、非特异性反应等多项指标进行统计学分析。结果 与传统SPA法相比,自组装SPA具有抗体固定量大、反应速度快、频率下降明显、非特异性反应低等优点。结论 自组装SPA法具有比普通SPA法包被一致性更高的优点,可以作为新一代免疫传感器的抗体固定方法。

OBJECTIVE To discuss and establish a highly effective method to immobilize antibody(Ab) on the surface of immunosensor microarray. METHODS Common protein A (SPA) and self-assembled monolayers SPA (SAMs SPA) were used to immobilize the antibody on the microarray respectively. Then different concentration of antibody and antigen conjugated on the quarts crystal microbalance(QCM) to compare the difference of the quantity of antibody immobilized, the time spent in the process of Ag-Ab complex formed and the nonspecific reaction between unrelated antigen. RESULTS Modified SPA had the advantage of more antibody immobilization, less time consumed in testing and lower nonspecific reaction. CONCLUSIONS SAMs SPA have better ability to immobilize antibody and can be applied to immobilize antibody on the surface of a new generation immunosensor.