Electrochemical immunoassay for carcinoembryonic antigen using gold nanoparticle-graphene composite modified glassy carbon electrode.

Carcinoembryonic antigen (CEA), which is typically associated with certain tumors and developing fetus, is widely used as a clinical tumor marker for some familiar cancers. In this work, a simple and sensitive electrochemical CEA sensor was developed by employing immunoreaction. Gold nanoparticle-decorated graphene composites (Au-GN) were successfully synthesized based on the reduction of HAuCl4 in the presence of graphene. Horseradish peroxidase-labeled anti-CEA antibody (HRP-anti-CEA) and HRP were successively adsorbed on the Au-GN modified glassy carbon electrode. The stepwise assembly process of the immunosensor was characterized by cyclic voltammetry and electrochemical impedance spectroscopy. The introduction of CEA antigens on the electrode surface reduced the electrochemical response of the electron transfer mediator due to the strong steric effect. Under the optimized conditions, the peak current change derived from the differential pulse voltammetry (DPV) measurements (ΔIDPV) was proportional to the CEA concentration from 0.10 to 80 ng mL(-1) with a detection limit of 0.04 ng mL(-1) (S/N=3). In addition, this new protocol shows good selectivity, stability and reproducibility. The determination of CEA in human serum samples was performed and received in excellent accordance with the results determined by the enzyme-linked immunosorbent assay (ELISA).

[1]  Andre K. Geim,et al.  The rise of graphene. , 2007, Nature materials.

[2]  R. F. Dutra,et al.  A dual quartz crystal microbalance for human cardiac troponin T in real time detection , 2012 .

[3]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[4]  D. Chan,et al.  Immunoassay : a practical guide , 1987 .

[5]  Jing Li,et al.  High-sensitivity determination of lead and cadmium based on the Nafion-graphene composite film. , 2009, Analytica chimica acta.

[6]  Concepción Martín,et al.  CEA and CA 549 in serum and pleural fluid of patients with pleural effusion. , 2002, Lung cancer.

[7]  Feng Yan,et al.  Novel amperometric immunosensor for rapid separation-free immunoassay of carcinoembryonic antigen. , 2004, Journal of immunological methods.

[8]  Rajendra N. Goyal,et al.  Differential pulse voltammetric determination of atenolol in pharmaceutical formulations and urine using nanogold modified indium tin oxide electrode , 2006 .

[9]  Kian Ping Loh,et al.  Optimizing label-free DNA electrical detection on graphene platform. , 2011, Analytical chemistry.

[10]  E. Eppler,et al.  Carcinoembryonic antigen (CEA) presentation and specific T cell-priming by human dendritic cells transfected with CEA-mRNA. , 2002, European journal of cancer.

[11]  Dan Wu,et al.  Label-free electrochemical immunosensor for prostate-specific antigen based on silver hybridized mesoporous silica nanoparticles. , 2013, Analytical biochemistry.

[12]  Xian‐Wen Wei,et al.  Fabrication of metal-graphene hybrid materials by electroless deposition , 2011 .

[13]  Dan Du,et al.  Functionalized graphene oxide as a nanocarrier in a multienzyme labeling amplification strategy for ultrasensitive electrochemical immunoassay of phosphorylated p53 (S392). , 2011, Analytical chemistry.

[14]  S. Novaković,et al.  Clinical evaluation of potential usefulness of CEA, CA 15-3, and MCA in follow-up of breast cancer patients. , 1996, Cancer letters.

[15]  H. Hollema,et al.  Serum CA 125, carcinoembryonic antigen, and CA 19-9 as tumor markers in borderline ovarian tumors. , 2000, Gynecologic oncology.

[16]  Chunxiang Xu,et al.  ZnO quantum dot labeled immunosensor for carbohydrate antigen 19-9. , 2011, Biosensors & bioelectronics.

[17]  Wei Wang,et al.  A disposable electrochemical immunosensor for carcinoembryonic antigen based on nano-Au/multi-walled carbon nanotubes-chitosans nanocomposite film modified glassy carbon electrode. , 2010, Analytica chimica acta.

[18]  Y. Chai,et al.  Enhancement of carcinoembryonic antibody immobilization on gold electrode modified by gold nanoparticles and SiO2/Thionine nanocomposite , 2009 .

[19]  Jianbin Zheng,et al.  Sensitive label-free electrochemical immunoassay based on a redox matrix of gold nanoparticles/Azure І/multi-wall carbon nanotubes composite , 2011 .

[20]  Chen-zhong Li,et al.  Detection and discrimination of alpha-fetoprotein with a label-free electrochemical impedance spectroscopy biosensor array based on lectin functionalized carbon nanotubes. , 2013, Talanta.

[21]  J. Sørensen,et al.  Carcinoembryonic antigen (CEA) as tumor marker in lung cancer. , 2012, Lung cancer.

[22]  Ying Zhuo,et al.  Amperometric immunosensor for simultaneous detection of three analytes in one interface using dual functionalized graphene sheets integrated with redox-probes as tracer matrixes. , 2013, Biosensors & bioelectronics.

[23]  Xi Zhang,et al.  Unconventional layer-by-layer assembly of graphene multilayer films for enzyme-based glucose and maltose biosensing. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[24]  Bing Zhang,et al.  Electrochemical immunosensor for carcinoembryonic antigen based on nanosilver-coated magnetic beads and gold-graphene nanolabels. , 2012, Talanta.

[25]  S. Yao,et al.  Direct electrochemistry of cholesterol oxidase immobilized on gold nanoparticles-decorated multiwalled carbon nanotubes and cholesterol sensing. , 2013, Talanta.

[26]  M. D. Rooij,et al.  Electrochemical Methods: Fundamentals and Applications , 2003 .

[27]  U. Bilitewski,et al.  Can affinity sensors be used to detect food contaminants? , 2000, Analytical chemistry.

[28]  Yunhui Yang,et al.  The preparation of label-free electrochemical immunosensor based on the Pt-Au alloy nanotube array for detection of human chorionic gonadotrophin. , 2011, Clinica chimica acta; international journal of clinical chemistry.

[29]  Dianping Tang,et al.  In situ amplified electrochemical immunoassay for carcinoembryonic antigen using horseradish peroxidase-encapsulated nanogold hollow microspheres as labels. , 2008, Analytical chemistry.

[30]  J. Kysar,et al.  Measurement of the Elastic Properties and Intrinsic Strength of Monolayer Graphene , 2008, Science.

[31]  Zhimin Zhang,et al.  Nanogold-enwrapped graphene nanocomposites as trace labels for sensitivity enhancement of electrochemical immunosensors in clinical immunoassays: Carcinoembryonic antigen as a model. , 2010, Biosensors & bioelectronics.

[32]  J. J. Gracio,et al.  Surface Modification of Graphene Nanosheets with Gold Nanoparticles: The Role of Oxygen Moieties at Graphene Surface on Gold Nucleation and Growth , 2009 .

[33]  Andrey V Lisitsa,et al.  Highly sensitive detection of human cardiac myoglobin using a reverse sandwich immunoassay with a gold nanoparticle-enhanced surface plasmon resonance biosensor. , 2013, Analytica chimica acta.

[34]  Dan Wu,et al.  Electrochemical immunoassay for carcinoembryonic antigen based on signal amplification strategy of nanotubular mesoporous PdCu alloy. , 2012, Biosensors & bioelectronics.

[35]  Faraggi,et al.  Methodological issues associated with tumor marker development. Biostatistical aspects. , 2000, Urologic oncology.

[36]  Jie Wu,et al.  Multilayer hemin/G-quadruplex wrapped gold nanoparticles as tag for ultrasensitive multiplex immunoassay by chemiluminescence imaging. , 2013, Biosensors & bioelectronics.

[37]  Jing‐Juan Xu,et al.  A novel lable-free electrochemical immunosensor for carcinoembryonic antigen based on gold nanoparticles-thionine-reduced graphene oxide nanocomposite film modified glassy carbon electrode. , 2011, Talanta.

[38]  Jinghua Yu,et al.  Ultrasensitive electrochemical immunoassay for carcinoembryonic antigen based on three-dimensional macroporous gold nanoparticles/graphene composite platform and multienzyme functionalized nanoporous silver label. , 2013, Analytica chimica acta.

[39]  Stela Pruneanu,et al.  Novel Graphene-Gold Nanoparticle Modified Electrodes for the High Sensitivity Electrochemical Spectroscopy Detection and Analysis of Carbamazepine , 2011 .

[40]  Jeanette M. Van Emon,et al.  IMMUNOCHEMICAL METHODS for Environmental Analysis , 1989 .

[41]  Yanbo Zeng,et al.  Graphene oxide as nanocarrier for sensitive electrochemical immunoassay of clenbuterol based on labeling amplification strategy. , 2013, Talanta.

[42]  Yanming Liu,et al.  Capillary electrophoresis immunoassay chemiluminescence detection of zeptomoles of bone morphogenic protein-2 in rat vascular smooth muscle cells. , 2004, Analytical chemistry.

[43]  Shifeng Hou,et al.  Highly Sensitive and Selective Dopamine Biosensor Fabricated with Silanized Graphene , 2010 .

[44]  P. A. Johnson,et al.  Surface-enhanced Raman scattering (SERS) detection of multiple viral antigens using magnetic capture of SERS-active nanoparticles. , 2013, Biosensors & bioelectronics.

[45]  Shen-ming Chen,et al.  A simple electrochemical approach to fabricate a glucose biosensor based on graphene-glucose oxidase biocomposite. , 2013, Biosensors & bioelectronics.