Abstract
The authors describe an electrochemical immunoassay for the core antigen of hepatitis C virus (HCV). The method is based on the use of a screen-printed carbon electrode (SPCE) that was modified with a Nafion@TiO2 nanocomposite and loaded with secondary antibody (Ab2) to entrap Celestine Blue (CB). The material has architecture of the type CB/Ab2/Nafion@TiO2. A nanocomposite consisting of graphene, ionic liquid and fullerene was deposited on the SPCE first, and rhodium nanoparticles (RhNPs) were then deposited on the surface of modified electrode in order to immobilize primary antibody (Ab1). The antigen and CB/Ab2/Nafion@TiO2 were conjugated one by one to form a sandwich-type immunocomplex. The signal was obtained by differential pulse voltammetry whose intensity is related to the concentration of the antigen. The assay, if operated at a working voltage of typically 0.35 V (vs. Ag/AgCl) has a response that is linear in the 0.1 to 250 pg⋅mL−1 HCV core antigen concentration range, and the limit of detection is as low as 25 fg⋅mL−1. The assay was applied to the determination of the HCV core antigen in spiked human serum samples. In our perception, the method represents a promising platform for the detection of various antigens if appropriate antibodies are available.
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References
Ergünay K, Şener B, Alp A, Karakaya J, Hasçelik G (2011) Utility of a commercial quantitative hepatitis C virus core antigen assay in a diagnostic laboratory setting. Diagn Microbiol Infect Dis 70:486–491
Ma C, Liang M, Wang L, Xiang H, Jiang Y, Li Y, Xie G (2013) Multis HRP-DNA-coated CMWNTs as signal labels for an ultrasensitive hepatitis C virus core antigen electrochemical immunosensor. Biosens Bioelectron 47:467–474
Icardi G, Ansaldi F, Bruzzone BM, Durando P, Lee S, De C, Luigi C (2001) Novel approach to reduce the hepatitis C virus (HCV) window period: clinical evaluation of a new enzyme-linked immunosorbent assay for HCV Core antigen. J Clin Microbiol 39:3110–3114
Leary TP, Gutierrez RA, Muerhoff AS, Birkenmeyer LG, Desai SM, Dawson GJ (2006) A chemiluminescent, magnetic particle-based immunoassay for the detection of hepatitis C virus core antigen in human serum or plasma. J Med Virol 78:1436–1440
Morota K, Fijinami R, Kinukawa H, Machida T, Ohno K, Saegusa H (2009) A new sensitive and automated chemiluminescent microparticle immunoassay for quantitative determination of hepatitis C virus core antigen. J Virol Methods 157:8–14
Tanaka E, Ohue C, Aoyagi K, Yamaguchi K, Yagi S, Kiyosawa K, Alter HJ (2000) Evaluation of a new enzyme immunoassay for hepatitis C virus (HCV) core antigen with clinical sensitivity approximating that of genomic amplification of HCV RNA. Hepatology 32:388–393
Valcavi P, Medici MC, Casula F, Arcangeletti MC, De Conto F, Pinardi F (2004) Evaluation of a total hepatitis C virus (HCV) core antigen assay for the detection of antigenaemia in anti-HCV positive individuals. J Med Virol 73:397–403
Okuno J, Maehashi K, Kerman K, Takamura Y, Matsumoto K, Tamiya E (2007) Label-free immunosensor for prostate-specific antigen based on single-walled carbon nanotube array-modified microelectrodes. Biosens Bioelectron 22:2377–2381
Salimi A, Kavosi B, Fathi F, Hallaj R (2013) Highly sensitive immunosensing of prostate-specific antigen based on ionic liquid–carbon nanotubes modified electrode: application as cancer biomarker for prostate biopsies. Biosens Bioelectron 42:439–446
Liu Y, Guo W, Qin X, Meng X, Zhu X, Wang J, Pei M, Wang L (2014) Sensitive sandwich electrochemical immunosensor for human chorionic gonadotropin using nanoporous Pd as a label. RSC Adv 4:21891–21898
Jeong B, Akter R, Han OH, Rhee CK, Rahman MA (2013) Increased Electrocatalyzed performance through Dendrimer-encapsulated gold nanoparticles and carbon nanotube-assisted multiple Bienzymatic labels: Highly sensitive electrochemical Immunosensor for protein detection. Anal Chem 85:1784–1791
Roushani M, Valipour A (2016) Voltammetric immunosensor for human chorionic gonadotropin using a glassy carbon electrode modified with silver nanoparticles and a nanocomposite composed of graphene, chitosan and ionic liquid, and using riboflavin as a redox probe. Microchim Acta 183:845–853
Hu C, Yang DP, Wang Z, Huang P, Wang X, Chen D, Cui D, Yang M, Jia N (2013) Bio-mimetically synthesized Ag@BSA microspheres as a novel electrochemical biosensing interface for sensitive detection of tumor cells. Biosens Bioelectron 41:656–661
Zhang Y, Zhang B, Ye X, Yan Y, Huang L, Jiang Z, Tan S, Cai X (2016) Electrochemical immunosensor for interferon-γ based on disposable ITO detector and HRP-antibody-conjugated nano gold as signal tag. Mater Sci Eng C 59:577–584
Roushani M, Valipour A (2016) Using electrochemical oxidation of Rutin in modeling a novel andsensitive immunosensor based on Pt nanoparticle and graphene–ionicliquid–chitosan nanocomposite to detect human chorionicgonadotropin. Sensors Actuators B Chem 222:1103–1111
Li R, Wu D, Li H, Xu C, Wang H, Zhao Y, Cai Y, Wei Q, Du B (2011) Label-free amperometric immunosensor for the detection of human serum chorionic gonadotropin based on nanoporous gold and graphene. Anal Biochem 414:196–201
Wei Q, Li R, Du B, Wu D, Han Y, Cai Y, Zhao Y, Xin X, Li H, Yang M (2011) Multifunctional mesoporous silica nanoparticles as sensitive labels for immunoassay of human chorionic gonadotropin. Sensors Actuators B Chem 153:256–260
Wu D, Zhang Y, Shi L, Cai Y, Ma H, Du B, Wei Q (2013) Electrochemical Immunosensor for ultrasensitive detection of human chorionic gonadotropin based on Pd@SBA-15. Electroanalysis 25:427–432
Liu Y, Wang G, Li C, Zhou Q, Wang M, Yang L (2014) A novel acetylcholinesterase biosensor based on carboxylic graphene coated with silver nanoparticles for pesticide detection. Mater Sci Eng C 35:253–258
Luo YP, Liu HQ, Rui Q, Tian Y (2009) Detection of extracellular H2O2 released from human liver cancer cells based on TiO2 Nanoneedles with enhanced electron transfer of cytochrome c. Anal Chem 81:3035–3041
Tang H, Yan F, Tai QD, Chan HLW (2010) The improvement of glucose bioelectrocatalytic properties of platinum electrodes modified with electrospun TiO2 nanofibers. Biosens Bioelectron 25:1646–1651
Xi F, Zhao D, Wang X, Chen P, Feng NX, Dong JZ, Xue WW, Peng C (2013) Non-enzymatic detection of hydrogen peroxide using a functionalized three-dimensional graphene electrode. Electrochem Commun 26:81–84
Er E, Celikkan H, Erk N (2016) Highly sensitive and selective electrochemical sensor based on high-quality graphene/nafion nanocomposite for voltammetric determination of nebivolol. Sensors Actuators B Chem 224:170–177
Mao L, Yuana R, Chai Y, Zhuo Y, Yang X (2010) A new electrochemiluminescence immunosensor based on Ru(bpy)3 2+-doped TiO2 nanoparticles labeling for ultrasensitive detection of human chorionic gonadotrophin. Sensors Actuators B Chem 149:226–232
Ma C, Xie G, Zhang W, Liang M, Liu B, Xiang H (2012) Label-free sandwich type of immunosensor for hepatitis C virus core antigen based on the use of gold nanoparticles on a nanostructured metal oxide surface. Microchim Acta 178:331–340
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This study was supported by the Foundation of Ilam University. Authors express their deep thanks.
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Valipour, A., Roushani, M. TiO2 nanoparticles doped with Celestine Blue as a label in a sandwich immunoassay for the hepatitis C virus core antigen using a screen printed electrode. Microchim Acta 184, 2015–2022 (2017). https://doi.org/10.1007/s00604-017-2190-7
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DOI: https://doi.org/10.1007/s00604-017-2190-7