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    基于功能化石墨纳米片的库仑型免疫传感器用于检测前列腺特异性抗原

    Detection of Prostate Specific Antigen by Coulometric Immunosensor Based on Functional Graphite Nano Sheets

    • 摘要: 以石墨为原料,1-芘丁酸(PBA)为表面活性剂,采用超声剥离法合成了PBA非共价功能化的石墨纳米片(PBA@GNSs),并采用扫描电子显微镜(SEM)对其进行表征。将PBA@GNSs撒在刷好的丝网印刷碳电极(SPCEs)表面,以实现电极的批量化修饰。用活化试剂N-羟基琥珀酰亚胺和1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(NHS-EDC)活化PBA的羧基,以牛血清白蛋白(BSA)为封闭剂,将PBA@GNSs固定抗体(Ab1),构建了Ab1/PBA@GNSs/SPCEs免疫传感器。采用循环伏安法和电化学交流阻抗谱法研究了上述传感器的构建过程,并用库仑法对前列腺特异性抗原(PSA)进行定量测定。结果表明,该免疫传感器的线性范围为10~3 000 ng·L-1,检出限为3.3 ng·L-1。对样品平行测定5次,测定值的相对标准偏差均小于2.0%,加标回收率为96.1%~101%。选择性和稳定性试验表明,该免疫传感器对PSA具有良好的选择性,且于4 ℃保存18 d后,库仑响应信号仅衰减为初始信号的92.0%。

       

      Abstract: Using graphite as raw material and 1-pyrene butyric acid (PBA) as surfactant, non covalently functionalized graphite nano sheets loaded with PBA (PBA@GNSs) were synthesized by ultrasonic stripping method, and characterized by scanning electron microscope(SEM). Then PBA@GNSs were sprinkled on the surface of brushed screen-printed carbon electrodes (SPCEs) to achieve batch modification of electrodes. Then the carboxyl group of PBA was activated by N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (NHS-EDC), bovine serum albumin (BSA) was used as the blocking agent, and antibody (Ab1) was fixed with PBA@GNSs to construct Ab1/PBA@GNSs/SPCEs immunosensor. Cyclic voltammetry and electrochemical impedance spectroscopy were used to study the construction process of the above sensor, and coulometry was used to quantify the prostate specific antigen (PSA). As shown by the results, the linear range of this immunosensor was 10-3 000 ng · L-1, with detection limit of 3.3 ng · L-1. Five parallel determinations were conducted on the samples, with RSDs of the determined values less than 2.0%. The recovery of the spiked samples was in the range of 96.1%-101%. It was shown by the selectivity and stability tests that the immunosensor had good selectivity for PSA, and after being stored at 4 ℃ for 18 d, the Coulomb response signal only decaied to 92.0% of the initial signal.

       

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