16-SPCEAmperometric PSA Immunosensor Modified by Gold Nanoparticles Electrodeposition
1 University of Shanghai for Science and Technology, Shanghai 200093, China
2 Shanghai Advanced Research Institute, Chinese Academy of Sciences, Shanghai 201210, China
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 screenprinted electrode (16SPCE) modified by gold nanoparticles electrodeposition. The surface of 16SPCE was modified by gold nanoparticles which was reduced by HAuCl 4 through potentiostatic deposition. HAuCl 4 solution with concentration of 005 mg/mL and deposition time of 150 s were the best condition for the 16SPCE 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 IT 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 screenprinted 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.
［1］Wang MC, Valenzuela LA, Murphy GP,et al Purification of a human prostate specific antigen［J］. Investigative Urology, 1979, 17(2): 159-163.
［2］Catalona WJ, Richie JP, Ahmann FR, et al. Comparison of digital rectal examination and serum prostate specific antigen in the early detection of prostate cancer: results of a multicenter clinical trial of 6,630 men［J］. The Journal of Urology, 1994, 151(5): 1283-1290.
［3］Becker C, Lilja H. Individal prostatespecific antigen (PSA) forms as prostate tumor markers［J］. Clinica Chimica Acta, 1997, 257: 117-132.
［4］MacKness BC, Chourb S, Farris LR, et al. Polymerproteinenhanced fluoroimmunoassay for prostatespecific antigen［J］. Analytical and Bioanalytical Chemistry,2010,396(2):681-686.
［5］胡萍，司同国，于海鹏，等. 联合检测血清HMGB1 PSA在局限性前列腺癌冷冻术后复发中的应用价值［J］. 中国肿瘤临床. 2013，40(8): 462-465.
［6］Seto Y, Iba T, Abe K. Development of ultrahigh sensitivity bioluminescent enzyme immunoassay for prostatespecific antigen (PSA) using firefly luciferase［J］. Luminescence, 2001, 16(4): 285-290.
［7］Xu Shoujiang, Liu Yang, Wang Taihong, et al. Positive potential operation of a cathodic electrogenerated chemiluminescence immunosensor based on luminol and graphene for cancer biomarker detection［J］. Analytical Chemistry, 2011, 83(10): 3817-3823.
［8］朱岚，黄飚，张祥瑞. 双标记前列腺特异性抗原时间分辨荧光免疫分析试剂盒研究［J］. 中华检验医学杂志, 2006，29(11): 1001-1004.
［9］Kokko T, Liljenback T, Peltola MT, et al. Homogeneous dualparameter assay for prostatespecific antigen based on fluorescence resonance energy transfer［J］. Analytical Chemistry, 2008, 80(24): 9763-9768.
［10］Ling Huiling, Lv Jingzhang, Zhu Zhihao, et al. New progress in detection of foodborne pathogenic Escherichia coli O157: H7 based on biosensors［J］. Journal of Food Safety and Quality, 2013, 4(6): 1828-1834.
［11］Dou Wenchao, Tang Weilu, Zhao Guangying, et al. A disposable electrochemical immunosensor arrays using 4channel screenprinted carbon electrode for simultaneous detection of Escherichia coli O157:H7 and Enterobacter sakazakii［J］. Electrochimica Acta,2013,97:79-85.
［12］Liu Fang, Deng Wenping, Zhang Yan, et al. Application of ZnO quantum dots dotted carbon nanotube for sensitive electrochemiluminescence immunoassay based on simply electrochemical reduced Pt/Au alloy and a disposable device［J］. Analytica Chimica Acta, 2014, 818: 46-53.
［13］Moon JM, Hui KY, Cho Y. A nanowirebased labelfree immunosensor: Direct incorporation of a PSA antibody in electropolymerized polypyrrole［J］. Biosensors and Bioelectronics, 2014, 57: 157-161.
［14］Kavosi B, Hallaj R, Teymourian H, et al. Au nanoparticles/PAMAM dendrimer functionalized wired ethyleneamine–viologen as highly efficient interface for ultrasensitive αfetoprotein electrochemical immunosensor［J］. Biosensors and Bioelectronics, 2014, 59: 389-396.
［15］许海楠,高鹏,王军，等. PSA及其相关指标与前列腺癌的诊断［J］.医学综述, 2010,16(12):1823-1825.
［16］Wong LS, Khan F, Micklefield J. Selective covalent protein immobilization: strategies and applications［J］. Chemical Reviews, 2009, 109: 4025-4053.
［17］Redeker ES, Ta DT, Cortens D, et al. Protein engineering for directed immobilization［J］. Bioconjugate Chemistry, 2013, 24: 1761-1777.
［18］Shao Weixie, Hong Lili, Jing Du, et al. Influence of serum prostatespecific antigen (psa) level, prostate volume, and psa density on prostate cancer detection with contrastenhanced sonography using contrasttuned imaging technology ［J］. J Ultrasound Med, 2013, 32:741-748.
