An electrochemical affinity sensor for the determination of fumonisins mycotoxins (Fms) using monoclonal antibody modified screen-printed gold electrode with carbon counter and silver-silver chloride pseudo-reference electrode is reported in this work. A direct competitive enzyme-linked immunosorbent assay (ELISA) was initially developed, exhibiting a detection limit of 100 μg·L-1for fumonisins. This was then transferred to the surface of a bare gold screen-printed electrode (SPGE) and detection was performed by chronoamperometry, monitoring the reaction of 3,3’,5,5’-Tetramethylbenzidine dihydrochloride (TMB) and hydrogen peroxide (H2O2) catalysed by HRP at ？100 mV potential vs. onboard Ag-AgCl pseudo-reference electrode. The immunosensor exhibited detection limit of 5 μg·L？1 fumonisins with a dynamic range from 1 μg·L？1–1000 μg·L？1. The sensor also performed well in extracted corn samples.
References
[1]
Gelderblom, W.C.A.; Jaskiewicz, K.; Marasas, W.F.O.; Thiel, P.G.; Horak, R.M.; Vleggaar, R.; Kriek, N.P.J. Fumonisins-Novel mycotoxins with cancer-promoting activity produced by Fusarium moniliforme. Appl. Environ. Microbiol.？1988, 54, 1806–1811. 2901247
[2]
Dutton, M.F. Fumonisins, mycotoxin of increasing importance: Their nature and their effects. Phramatcol. Ther.？1996, 70, 137–161, doi:10.1016/0163-7258(96)00006-X.
[3]
Shepard, G.S.; Thiel, P.G.; Stockenstorom, S.; Sydenham, E.W. Worldwide survey of fumonisin contamination of corn and corn-based products. J. AOAC Int.？1996, 79, 671–787. 8634537
[4]
Rheeder, J.P.; Marasas, W.F.O.; Vismer, H.F. Production of fumonisin analogs by Fusarium species. Appl. Environ. Microbiol.？2002, 68, 2101–2105. 11976077
[5]
Jackson, L.; Jablonski, J. Fumonisins. In Mycotoxins in Food, Detection and Control; Magan, N., Olsen, M., Eds.; Woodhead Publishing Ltd.: Cambridge, UK, 2004; pp. 367–405.
[6]
Marasas, W.F.O. Discovery and occurrence of the fumonisin: A historical perspective. Environ. Health Perspect.？2001, 109, 239–243. 11359691
[7]
Colvin, B.M.; Harrison, L.R. Fumonisins induced pulmonary edema and hydrothorax in swine. Mycopathologia？1992, 117, 79–82, doi:10.1007/BF00497282. 1513376
[8]
Lino, C.M.; Silva, L.J.G.; Pena, A.L.S.; Silveira, M.I. Determination of fumonisins B1 and B2 in Portuguese maize and maize-based samples by HPLC with fluorescence detection. Anal. Bioanal. Chem.？2006, 384, 1214–1220. 16465502
[9]
Rheeder, J.P.; Marasas, W.F.O.; Thiel, P.G.; Sydenham, E.W.; Sherpard, G.S.; van Schalkwyk, D.J. Fusarium moniliforme and fumonisins in corn in relation to human esophagaea cancer in Transkei. Phytopathology？1992, 82, 353–357, doi:10.1094/Phyto-82-353.
[10]
Pagliuca, G.; Zironi, E.; Ceccolini, A.; Matera, R.; Serrazanetti, P.G.; Piva, A. Simple method for the simultaneous isolation and determination of fumonisin B1 and its metabolite aminopentol-1 in swine liver by liquid chromatography–fluorescence detection. J. Chrom. B？2005, 819, 97–103, doi:10.1016/j.jchromb.2005.01.035.
[11]
Wang, S.; Quan, Y.; Lee, N.; Kennedy, R.I. Rapid determination of fumonisin B1 in food samples by enzyme-linked immunosorbent assay and colloid gold immunoassay. J. Agric. Food Chem.？2006, 54, 2491–2495. 16569033
[12]
Shelby, R.A.; Rottinghaus, G.E.; Minor, H.C. Comparison of thin layer chromatography and competitive immunoassay methods for detecting fumonisins on maize. J. Agric. Food Chem.？1994, 42, 2064–2067, doi:10.1021/jf00045a043.
[13]
Sydenham, E.W.; Shephard, G.S. Chromatographic and allied methods of analysisfor selected mycotoxins. In Progress in Food Contaminant Analysis; Gilbert, J., Ed.; Blackie Academic & Professional: London, UK, 1996; pp. 65–146.
[14]
Shepard, G.S. Chromatographic determination of the fumonisins mycotoxins. J. Chrom. A？1998, 815, 31–39, doi:10.1016/S0021-9673(98)00187-3.
[15]
Schneider, E.; Usleber, E.; M？rtlbauer, E. Rapid detection of fumonisin B1 in corn-based food by competitive direct dipstick enzyme immunoassay/enzyme-linked immunofiltration assay with integrated negative control reaction. J. Agric. Food Chem.？1995, 43, 2548–2552, doi:10.1021/jf00057a042.
[16]
Collin, R.; Schneider, E.; Briggs, L.; Towers, N. Development of immunodiagnostic field tests for the detection of the mycotoxin, sporidesmin A. Food Agric. Immunol.？1998, 10, 91–104, doi:10.1080/09540109809354973.
[17]
Sibanda, L.; De Saeger, S.; Van Peteghem, C.; Grabarkiewicz-Szczesna, J.; Tomczak, M. Detection of T-2 toxin in different cereals by flow-through enzyme immunoassay with a simultaneous internal reference. J. Agric. Food Chem.？2000, 48, 5864–5867. 11141260
[18]
Mullett, W.; Lai, E.P.; Yeung, J.M. Immunoassay of fumonisins by a surface plasmon resonance biosensor. Anal. Biochem.？1998, 258, 161–167. 9570825
[19]
Logrieco, A.; Arrigan, D.W.M.; Brengel-Pesce, K.; Siciliano, P.; Tothill, I.E. DNA arrays, electronic noses and tongues, biosensors and receptors for rapid detection of toxigenic fungi and mycotoxins: A review. Food Addit. Cont.？2005, 22, 335–344, doi:10.1080/02652030500070176.
[20]
Tothill, I.E. Biosensors developments and potential applications in the agricultural diagnosis sector. Comp. Elect. Agric.？2001, 30, 205–218, doi:10.1016/S0168-1699(00)00165-4.
[21]
Gaag, V.D.B.; Spath, S.; Dietrich, H.; Stigter, E.; Boonzaaijer, G.; van Osenbruggen, T.; Koopal, K. Biosensor and multiple mycotoxin analysis. Food Control？2003, 14, 251–254, doi:10.1016/S0956-7135(03)00008-2.
[22]
Daly, S.J.; Keating, G.J.; Dillon, P.P.; Manning, B.M.; O’Kennedy, R.; Lee, H.A.; Morgan, R.A. Development of surface plasmon resonance-based immunoassay for aflatoxin B1. J. Agric. Food Chem.？2000, 48, 5097–5104. 11087443
[23]
Thompson, V.S.; Maragos, C.M. Fiber–optic Immunosensor for the detection of fumonisin B1. J. Agric. Food Chem.？1996, 44, 1041–1046, doi:10.1021/jf950661t.
[24]
Tothill, I.E. On-line immunochemical assays for contaminant analysis. In Rapid and on-line Instrumentation for Food Quality Assurance; Tothill, I.E., Ed.; Woodhead Publishing Limited: Cambridge, UK, 2003; pp. 15–39.
[25]
Salam, F.; Tothill, I.E. Detection of Salmonella typhimurium using an electrochemical immunosenor. Biosen. Bioelec.？2009, 24, 2630–2636, doi:10.1016/j.bios.2009.01.025.
[26]
Parker, O.P.; Tothill, I.E. Development of an electrochemical immunosensor for aflatoxin M1 in milk wit focus on matrix interference. Biosen. Bioelec.？2009, 24, 2452–2457, doi:10.1016/j.bios.2008.12.021.
[27]
Crew, A.; Alford, C.; Cowell, D.C.C.; Hart, J.P. Development of a novel electrochemical immuno-assay using a screen printed electrode for the determination of secretory immunoglobulin a in human sweat. Electroch. Acta？2007, 52, 5232–5237, doi:10.1016/j.electacta.2007.02.035.
[28]
Butler, D.; Guilbault, G.G. Disposable amperometric immunosensor for the detection of 17-？ estradiol using screen-printed electrodes. Sens. Actuat. B？2006, 113, 692, doi:10.1016/j.snb.2005.07.019.
[29]
Micheli, L.; Grecco, R.; Badea, M.; Moscone, D.; Palleschi, G. An electrochemical immunosensor for aflatoxin M1 determination in milk using screen-printed electrodes. Biosens. Bioelec.？2005, 21, 588–596, doi:10.1016/j.bios.2004.12.017.
[30]
Studentsov, Y.Y.; Schiffman, M.; Strickler, H.D.; Ho, G.Y.F.; Susana Pang, Y.Y.; Schiller, J.; Herrero, R.; Burk, R.D. Enhanced enzyme-linked immunosorbent assay for detection of antibodies to virus-like particles of human papillomavirus. J. Clin. Microbil.？2002, 40, 1755–1760, doi:10.1128/JCM.40.5.1755-1760.2002.
[31]
Warwick, M.J. Chap 8: Standardisation of immunoassay. In Immunoassay, A Practical Guide; Brian, L., Ed.; Taylor & Francis Ltd.: London, UK, 1996; p. 160.
[32]
Ammida, N.H.S.; Micheli, L.; Palleschi, G. Electrochemical immunosensor for determination of aflatoxin B1 in barley. Analy. Chim. Acta？2004, 50, 159–164.
[33]
Alarcon, S.H.; Palleschi, G.; Compagnone, D.; Pascale, M.; Visconti, A.; Barna-Vetro, I. Monoclonal antibody based electrochemical immunosensor for the determination of ochratoxin A in wheat. Talanta？2006, 69, 1031–1037. 18970676
[34]
Volpe, G.; Compagnone, D.; Draisci, R.; Palleschi, G. 3,3,5,5-Tetramethylbenzidine as electrochemical substrate for horseradish peroxidase based enzyme immunoassays. A comparative study. Analyst？1998, 123, 1303–1307, doi:10.1039/a800255j.
[35]
Fu, X.H. Electrochemical measurement of DNA hybridization using nanosilver as label and horseradish peroxidase as enhancer. Bioprocess Biosyst. Eng.？2008, 31, 69–73, doi:10.1007/s00449-007-0147-7. 17671798
[36]
Muscarella, M.; Margo, L.S.; Nardiello, D.; Palermo, C.; Centonze, D. Development of a new analytical method for the determination of fumonisin B1 and B2 in food products based on high performance liquid chromatography and fluorimetric detection with post-column derivatization. J. Chrom. A.？2008, 1203, 88–93, doi:10.1016/j.chroma.2008.07.034.
[37]
Noh, M.F.M.; Tothill, I.E. Development and characterisation of disposable gold electrodes, and their use for lead (II) analysis. Anal. Bioanal. Chem.？2006, 386, 2095–2210, doi:10.1007/s00216-006-0904-5. 17086386
[38]
Anna, Y.K.; Shim, W.B.; Yang, Z.Y.; Eremin, S.A.; Chung, D.H. Direct competitive ELISA based on a monoclonal antibody for detection of aflatoxin B1. Stabilization of ELISA kit components and application to grain samples. Anal. Bioanal. Chem.？2006, 384, 286–294, doi:10.1007/s00216-005-0103-9. 16254721
