电沉积纳米金修饰的16通道电流型PSA免疫传感器的制备

doi:10.3969/j.issn.0258-8021.2015.01.008

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摘要近年来有关电化学免疫传感器的研究报道虽然很多，但普遍存在如预处理操作繁琐、灵敏度低、特异性差和成本昂贵等不足。本研究通过电沉积纳米金修饰16通道丝网印刷电极，以提高免疫传感器的性能。采用恒电位沉积法将氯金酸（HAuCl4）直接还原成纳米金，并修饰于16通道丝网印刷碳电极(16 SPCE)表面；以浓度为0.05 mg/mL的HAuCl4溶液和150 s电沉积时间为电沉积纳米金修饰16 SPCE的最优条件。利用静电吸附原理将前列腺抗原（PSA)抗体固定在电极表面，结合双抗夹心法制备免疫电极。通过循环伏安法和稳态时间电流曲线法表征免疫电极的性能。对30份前列腺癌患者和3份正常人血清进行PSA检测，与标准ELISA方法对照。结果表明：获得的免疫电极线性检测范围为0.2~100 ng/mL，其线性回归方程为Y=134.558X +72.705，线性相关系数为1，检测限为0.14 ng/mL。采用免疫传感器检测到的前列腺癌患者血清PSA水平与标准ELISA方法检测结果具有良好的线性相关性（R=0.944，P＜0.001）,正常人的检测值符合正常人血清中PSA的含量。所研制的免疫传感器具有灵敏度高、特异性强、成本低和反应时间短等特点，具有应用前景。

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	王泽华¹
	2 曾冬冬² 张欢² 宓现强^2*

关键词 ：纳米金, 电沉积, 16通道丝网印刷电极, 电化学免疫传感器, 前列腺特异性抗原

Abstract：Numerous studies have been reported about electrochemical immunosensor in recent years, while there still exist some shortcomings like complex preparation processes, low sensitivity, bad specificity and high cost. The purpose of this study is to improve the immunosensor performance using 16 channels screenprinted electrode (16SPCE) modified by gold nanoparticles electrodeposition. The surface of 16SPCE was modified by gold nanoparticles which was reduced by HAuCl 4 through potentiostatic deposition. HAuCl 4 solution with concentration of 005 mg/mL and deposition time of 150 s were the best condition for the 16SPCE modified by gold nanoparticles. Prostate antigen (PSA) antibody was fixed to the electrode surface based on electrostatic adsorption principle and PSA immunity electrode was made using double antibody sandwich method. The electrode performance was evaluated by cyclic voltammetry method and steady state IT curves method. The results of serum PSA levels detected by the immunosensor from 30 prostate cancer patient samples and 3 normal samples were consistent with those of ELISA method. The linear range of the immunesensor is from 0.2 ng/mL to 100 ng/mL, the linear regression equation is Y=134.558X+72.705, the linear correlation coefficient is 1 and the detection limit is 0.14 ng/mL. The results of serum PSA level from prostate cancer patient samples detected by the immunesensor show a good linear correlation with those of ELISA (R=0.944, P<0.001) and the results of serum PSA levels from normal samples are in the range of normal values. It means that the immunesensor based on 16 channel screenprinted electrode modified by gold nanoparticles has the advantages of high sensitivity, good specificity, low cost and short response time. The electrode shows promising clinical setting in the future.

Key words： gold noparticles electrodeposition 16 channel screen printed electrode electrochemical immunosensor prostate specific antigen

基金资助:上海市科委“创新行动计划”院市合作项目（13DZ1940200）；上海市青年科技英才扬帆计划(15YF1413300)

引用本文:

王泽华^1,2 曾冬冬² 张欢² 宓现强^2*. 电沉积纳米金修饰的16通道电流型PSA免疫传感器的制备[J]. 中国生物医学工程学报, 2015, 34(1): 55-61.
Wang Zehua^1,2Zeng Dongdong²Zhang Huan²Mi Xianqiang^2*. 16-SPCEAmperometric PSA Immunosensor Modified by Gold Nanoparticles Electrodeposition. journal1, 2015, 34(1): 55-61.

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http://cjbme.csbme.org/CN/10.3969/j.issn.0258-8021.2015.01.008 或 http://cjbme.csbme.org/CN/Y2015/V34/I1/55

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