16通道无标记型糖化血红蛋白检测免疫传感器的构建

Construction of a 16 channel label-free electrochemical immunosensor for glycated hemoglobin A1c detection

目的构建1种检测糖化血红蛋白(HbA1c)的无标记电化学免疫传感器。方法通过HbA1c抗体修饰16通道丝网印刷碳电极(16-SPCE),制备特异检测HbA1c的无标记电化学免疫传感器。采用循环伏安法测定16-SPCE电化学免疫传感器检测HbA1c的电信号,优化检测条件(HbA1c抗体最佳包被浓度、HbA1c抗原最佳孵育时间、电极的电化学特性、不同修饰电极的性能),同时进行初步方法学评价。结果HbA1c抗体最佳包被浓度为50μg/mL,HbA1c抗原最佳孵育时间为60 min。构建的16-SPCE电化学免疫传感器检测HbA1c的线性范围为1~50μg/mL,线性方程为Y=0.199X+10.468(r=0.955),检测限为0.88μg/mL。50μg/mL牛血清白蛋白(BSA)、50μg/mL前列腺特异性抗原(PSA)和50μg/mL葡萄糖对16-SPCE电化学免疫传感器检测HbA1c无干扰。制备的免疫电极在4℃能稳定放置5 d。采用16-SPCE电化学免疫传感器检测模拟血清样本中的HbA1c,各浓度的电流响应值能明显区分,且检测结果的重复性较好。结论构建的16-SPCE电化学免疫传感器具有良好的电化学响应能力、线性范围、抗干扰能力等。

Objective To construct a label-free electrochemical immunosensor for the specific detection of glycated hemoglobin A1c(HbA1c).Methods A 16 channel screen-printed carbon electrode(16-SPCE)was modified by HbA1c antibody to prepare a label-free electrochemical immunosensor for the specific detection of HbA1c.Cyclic voltammetry was used to detect the electrical signal of HbA1c by 16-SPCE electrochemical immunosensor,and the detection conditions(the optimal coating concentration of HbA1c antibody,the optimal incubation time of HbA1c antigen,the electrochemical characteristics of the electrode and the performance of different modified electrodes)were optimized.A preliminary methodological evaluation was conducted.Results The optimal HbA1c antibody coating concentration was 50μg/mL,and the optimal HbA1c antigen incubation time was 60 min.The linear range of HbA1c was 1-50μg/mL,and the linear equation was Y=0.199X+10.468(r=0.955).The detection limit was 0.88μg/mL,and 50μg/mL bovine serum albumin(BSA),50μg/mL prostate-specific antigen(PSA)and 50μg/mL glucose did not interfere with the detection of HbA1c by 16-SPCE electrochemical immunosensor.The prepared immunoelectrode was stably placed at 4°C for 5 d.The 16-SPCE electrochemical immunosensor was used to detect HbA1c in the simulated serum samples.The current response values of each concentration were clearly distinguished,and the repeatability was good.Conclusions The constructed 16-SPCE electrochemical immunosensor has good electrochemical response,linear range and anti-interference ability.