An ultrasensitive electrochemiluminescence immunosensor for alpha-fetoprotein based on a poly(aniline-luminol)/graphene oxide nanocomposite
- 34 Downloads
An ultrasensitive electrochemiluminescence (ECL) immunosensor for alpha-fetoprotein (AFP) detection based on a poly(aniline-luminol)/graphene oxide nanocomposite was developed. The nanocomposite, which was prepared using a fast and simple chemical oxidation strategy for the first time, showed excellent ECL performance. This outstanding ECL performance is due to the formation of poly(aniline-luminol) on the graphene oxide (GO) surface and the excellent electron-transfer properties of GO. Moreover, the poly(aniline-luminol)/graphene oxide nanocomposite has abundant amino groups at its surface, making it a good platform for biomacromolecule labeling. Using the nanocomposite, a novel ECL immunosensor for the determination of AFP was successfully developed. Anti-AFP was immobilized on the surface of a poly(aniline-luminol)/graphene oxide nanocomposite-modified electrode using a glutaraldehyde crosslinking method to form the ECL immunosensor. The AFP was then captured at the modified electrode surface through an antigen–antibody immunoreaction. When the AFP was captured by its antibody, the ECL intensity decreased. This ECL immunosensor for the detection of AFP exhibited a linear range of 1.7 × 10−12 to 1.7 × 10−8 mg mL−1 and a detection limit of 5 × 10−13 mg mL−1, indicating high sensitivity and linearity across a wide concentration range. Furthermore, the immunosensor was successfully applied to determine AFP in a real-world human serum sample.
KeywordsAlpha-fetoprotein Electrochemiluminescence Immunosensor Chemical oxidation Poly(aniline-luminol)/graphene oxide nanocomposite
This research was financially supported by the National Natural Science Foundation of China (no. 21665025), the Outstanding Young Science and Technology Personnel Training Project of the Xinjiang Uyghur Autonomous Region (no. QN2016YX0306), and Key Projects of the Engineering Research Center of Electrochemical Technology and Application of Xinjiang Normal Universitly (XJNUGCZX122017A01); all of this support is gratefully acknowledged.
Compliance with ethical standards
Ethics approval and consent to participate
All procedures performed in this study involving human participants were in accordance with the ethical standards of the 1964 Declaration of Helsinki and its later amendments, and were approved by the Ethical Committee of Xinjiang Normal University. All blood samples were from healthy persons, and informed consent was obtained from all individual participants included in the study.
Human and animal rights
No violation of human or animal rights occurred during this investigation.
Conflict of interest
The authors declare that there is no conflict of interest.
- 3.Yuan YL, Li SS, Xue YW, Liang JT, Cui LJ, Li QB, et al. A Fe3O4@Au-based pseudo-homogeneous electrochemical immunosensor for AFP measurement using AFP antibody-GNPs-HRP as detection probe. Anal Biochem. 2017;534:56–63.Google Scholar
- 10.Shen HY, Wang XQ, Wang YY, Qu F. Immunoassay for alpha fetal protein in serum based on quartz capillary materials by gray level analysis. Chin J Anal Chem. 2017;45:83–8.Google Scholar
- 14.Xiao LJ, Chai YQ, Wang HJ, Yuan R. Electrochemiluminescence immunosensor using poly(l-histidine)-protected glucose dehydrogenase on Pt/Au bimetallic nanoparticles to generate an in situ co-reactant. Analyst. 2014;139:4044–50.Google Scholar
- 16.Guo ZY, Sha YH, Hu YF, Yu ZQ, Tao YY, Wu YJ, et al. Faraday cage-type electrochemiluminescence immunosensor for ultrasensitive detection of Vibrio vulnificus based on multi-functionalized graphene oxide. Anal Bioanal Chem. 2016;408:7203–11.Google Scholar
- 20.Ferreira V, Cascalheira AC, Abrantes LM. Electrochemical preparation and characterisation of poly(luminol-aniline) films. Thin Solid Films. 2008;516:3996–4001.Google Scholar
- 22.Wang YJ, Li GX, Zheng XW. Electrochemiluminescence performance of poly(aniline-luminol) composite nanowires synthesized by chemical oxidation. Chin J Anal Chem. 2015;43:141–5.Google Scholar
- 23.Xiao CB, Zhao Q, Tu YF. An innovative route to prepare the all-solid-state electrochemiluminescent electrode—using the nano-rods of luminol/aniline copolymer. Electrochim Acta. 2014;147:791–6.Google Scholar
- 24.Zheng J, Ma XF, He XC, Gao MJ, Li G. Preparation, characterizations, and its potential applications of PANi/graphene oxide nanocomposite. Procedia Eng. 2012;27:1478–87.Google Scholar
- 25.Binh Phan T, Thanh Luong T, Mai TX, Thanh Thuy Mai T. Effect of nanostructured graphene oxide on electrochemical activity of its composite with polyaniline titanium dioxide. Adv Nat Sci-Nanosci. 2016;7:1–5.Google Scholar
- 26.Mohamed MA, Atty SA, Merey HA, Fattah TA, Foster CW, Banks CE. Titanium nanoparticles (TiO2)/graphene oxide nanosheets (GO): an electrochemical sensing platform for the sensitive and simultaneous determination of benzocaine in the presence of antipyrine. Analyst. 2017;142:3674–9.CrossRefGoogle Scholar
- 32.Gao K, Sun Z, Pan BG, Qiao XW, Hong CL. Electrochemical immunosensor for alpha-fetoprotein based on Prussian blue-carbon nanotube@polydopamine. Micro Nano Lett. 2018;13:58–62.Google Scholar
- 33.Li YK, Feng T, Deng SM, Wang XF, Xie MX. Development of magnetic and fluorescent immune sensors for the detection of alpha fetoprotein. Spectrosc Spectr Anal. 2017;37:1667–72.Google Scholar
- 36.Wang ZY, Guo WY, Di JW, Tu YF. The studies of electrochemical polymerization of luminol and the electrochemiluminescence properties on indium-tin oxide glass electrode. Chin J Anal Chem. 2005;33:763–6.Google Scholar
- 37.He S, Liang JM, Hu HL, Yuan JN, He JB, Ni ZF. Preparation of graphene oxide/chitosan layer-by-layer self-assembled composite film and its application in environmental protection. Mater Rev. 2016;30:1–5.Google Scholar
- 38.Yu M, Liu PR, Sun YJ, Liu JH, An JW, Li SM. Fabrication and characterization of graphene-Ag nanoparticles composites. J Inorg Mater. 2012;27:89–94.Google Scholar
- 39.Wang LL, Xing RG, Zhang BW, Hou Y. Preparation and electrochemical properties of functionalized graphene/polyaniline composite electrode materials. Acta Phys-Chim Sin. 2014;30:1659–66.Google Scholar