Electrochemical piezoelectric reusable immunosensor for aflatoxin B1 detection

Abstract A competitive, reusable, reliable and sensitive immunosensor based on electrochemical quartz crystal microbalance (EQCM) was developed for the detection of aflatoxin B1 (AFB1). The sensing platform was developed using a primary monoclonal anti-aflatoxin antibody (anti AFB1) covalently immobilized on a monolayer of 4-amino thiophenol self assembled on a gold coated quartz crystal electrode (anti AFB1/4-ATP/Au). The reusability was achieved by a sandwiched system using secondary rabbit-immunoglobulin antibodies (r-IgGs) tagged with core shell of gold coated iron oxide nanoparticles (r-IgG–Au-Fe3O4), which can be used to regenerate the bioelectrode. A competitive mode was employed, between free and coated AFB1 for fixed concentration of nanoparticle conjugate (r-IgG–Au–Fe3O4). After the competitive interaction, immunoelectrode was washed with phosphate saline buffer (PBS) at pH 7.4 and examined by electrochemical quartz crystal microbalance-cyclic voltammetry (EQCM-CV). Under the optimized conditions, the fabricated immunosensor can be used to detect AFB1 quantitatively with a linear range of 0.05 to 5 ng mL−1. For reliability, the fabricated immuno sensor was tested using cereal samples spiked with different concentrations of AFB1. In addition, this immuno electrode displays good reproducibility, and storage stability. The above immunosensor was regenerated with negligible loss in activity through removal of the r-IgG–Au–Fe3O4 conjugate using a strong magnet. It is shown that theAFB1/BSA/aAFB1/4-ATP/Au immuno sensor is a new approach for developing sensitive reliable and reusable electrochemical piezoelectric immunosensors.

[1]  A. Zuckerman,et al.  IARC Monographs on the Evaluation of Carcinogenic Risks to Humans , 1995, IARC monographs on the evaluation of carcinogenic risks to humans.

[2]  Xueping Liu,et al.  Biocatalyzed deposition amplification for detection of aflatoxin B1 based on quartz crystal microbalance. , 2009, Analytica chimica acta.

[3]  E. Levänen,et al.  Gas phase synthesis of encapsulated iron oxide–titanium dioxide composite nanoparticles by spray pyrolysis , 2013 .

[4]  Wenbin Liang,et al.  A novel magnetic Fe3O4@gold composite nanomaterial: Synthesis and application in regeneration-free immunosensor , 2010 .

[5]  D. Cliffel,et al.  Electrochemical sensors and biosensors. , 2012, Analytical chemistry.

[6]  A. Dasgupta,et al.  Iron oxide nanoparticles coated with gold: Enhanced magnetic moment due to interfacial effects , 2011 .

[7]  Lisa C Shriver-Lake,et al.  Indirect competitive immunoassay for detection of aflatoxin B1 in corn and nut products using the array biosensor. , 2006, Biosensors & bioelectronics.

[8]  Susana Campuzano,et al.  Preparation of core-shell Fe3O4@poly(dopamine) magnetic nanoparticles for biosensor construction. , 2014, Journal of materials chemistry. B.

[9]  R. Doong,et al.  Sensitive amperometric immunosensor for α-fetoprotein detection based on multifunctional dumbbell-like Au-Fe3O4 heterostructures , 2013 .

[10]  F. Lin,et al.  A real time detection of the ovarian tumor associated antigen 1 (OVTA 1) in human serum by quartz crystal microbalance immobilized with anti-OVTA 1 polyclonal chicken IgY antibodies. , 2012, Materials science & engineering. C, Materials for biological applications.

[11]  William W. Yu,et al.  Food related applications of magnetic iron oxide nanoparticles: Enzyme immobilization, protein purification, and food analysis , 2012 .

[12]  Tarasankar Pal,et al.  Magnetite nanoparticles with tunable gold or silver shell. , 2005, Journal of colloid and interface science.

[13]  S. Shen,et al.  Design and construction of polymerized-chitosan coated Fe3O4 magnetic nanoparticles and its application for hydrophobic drug delivery. , 2015, Materials science & engineering. C, Materials for biological applications.

[14]  Zhichuan J. Xu,et al.  Magnetic core/shell Fe3O4/Au and Fe3O4/Au/Ag nanoparticles with tunable plasmonic properties. , 2007, Journal of the American Chemical Society.

[15]  R. Chauhan,et al.  A novel electrochemical piezoelectric label free immunosensor for aflatoxin B1 detection in groundnut , 2015 .

[16]  S. Dmitrienko,et al.  Determination of cysteamine using label-free gold nanoparticles , 2012 .

[17]  E. Alocilja,et al.  Electrically active polyaniline coated magnetic (EAPM) nanoparticle as novel transducer in biosensor for detection of Bacillus anthracis spores in food samples. , 2009, Biosensors & bioelectronics.

[18]  Ashok Kumar Das,et al.  Preparation of sulfonated poly(ether–ether–ketone) functionalized ternary graphene/AuNPs/chitosan nanocomposite for efficient glucose biosensor , 2013 .

[19]  Guangming Zeng,et al.  Amplified and selective detection of manganese peroxidase genes based on enzyme-scaffolded-gold nanoclusters and mesoporous carbon nitride. , 2015, Biosensors & bioelectronics.

[20]  Ajeet Kaushik,et al.  Self-assembled monolayer based impedimetric platform for food borne mycotoxin detection. , 2010, Nanoscale.

[21]  S. Veintemillas-Verdaguer,et al.  Short-chain PEG molecules strongly bound to magnetic nanoparticle for MRI long circulating agents. , 2013, Acta biomaterialia.

[22]  Giuseppe Palleschi,et al.  Development of a QCM (Quartz Crystal Microbalance) Biosensor to the Detection of Aflatoxin B1 , 2013 .

[23]  Jian-hui Jiang,et al.  Piezoelectric immunosensor with gold nanoparticles enhanced competitive immunoreaction technique for quantification of aflatoxin B1. , 2009, Biosensors & bioelectronics.

[24]  Bin Du,et al.  Ultrasensitive electrochemical immunosensor for zeranol detection based on signal amplification strategy of nanoporous gold films and nano-montmorillonite as labels. , 2013, Analytica chimica acta.

[25]  Leila Masoomi,et al.  A non-enzymatic nanomagnetic electro-immunosensor for determination of Aflatoxin B1 as a model antigen , 2013 .

[26]  B. Saad,et al.  Micro-solid phase extraction with liquid chromatography-tandem mass spectrometry for the determination of aflatoxins in coffee and malt beverage. , 2014, Food chemistry.

[27]  I. Robinson,et al.  Synthesis of core-shell gold coated magnetic nanoparticles and their interaction with thiolated DNA. , 2010, Nanoscale.

[28]  Jaesung Jang,et al.  Mechanical desorption of immobilized proteins using carbon dioxide aerosols for reusable biosensors. , 2015, Analytica chimica acta.

[29]  P. Mukherjee,et al.  Synthesis and magnetic properties of gold coated iron oxide nanoparticles , 2009 .

[30]  Taeghwan Hyeon,et al.  Synthesis of monodisperse spherical nanocrystals. , 2007, Angewandte Chemie.

[31]  I. Karube,et al.  Sensitive detection of ochratoxin A in wine and cereals using fluorescence-based immunosensing. , 2012, Food chemistry.

[32]  Martin Hegner,et al.  Ultralarge atomically flat template-stripped Au surfaces for scanning probe microscopy , 1993 .

[33]  Development of Triglyceride Biosensor Based on a Platinum Nano Particle and Polypyrrole Nano Composite Electrode , 2013 .

[34]  D. Vanhecke,et al.  Preparation and characterization of functional silica hybrid magnetic nanoparticles , 2014 .

[35]  Huy L. Nguyen,et al.  Label-free detection of aflatoxin M1 with electrochemical Fe3O4/polyaniline-based aptasensor. , 2013, Materials science & engineering. C, Materials for biological applications.

[36]  Bo Bae Kim,et al.  Label-free homogeneous FRET immunoassay for the detection of mycotoxins that utilizes quenching of the intrinsic fluorescence of antibodies. , 2013, Biosensors & bioelectronics.

[37]  Guo-Li Shen,et al.  Piezoelectric immunosensor based on magnetic nanoparticles with simple immobilization procedures , 2003 .

[38]  Ronald W Davis,et al.  Sensitive giant magnetoresistive-based immunoassay for multiplex mycotoxin detection. , 2010, Biosensors & bioelectronics.

[39]  Y. Ying,et al.  A simple and rapid optical biosensor for detection of aflatoxin B1 based on competitive dispersion of gold nanorods. , 2013, Biosensors & bioelectronics.

[40]  Zhouping Wang,et al.  Magnetic nanobead-based immunoassay for the simultaneous detection of aflatoxin B1 and ochratoxin A using upconversion nanoparticles as multicolor labels. , 2011, Biosensors & bioelectronics.

[41]  Na Li,et al.  A reusable quartz crystal microbalance biosensor for highly specific detection of single-base DNA mutation. , 2013, Biosensors & bioelectronics.

[42]  B. Varughese,et al.  Magnetic iron oxide nanoparticles for biorecognition: evaluation of surface coverage and activity. , 2006, The journal of physical chemistry. B.

[43]  P. Solanki,et al.  Antibody immobilized cysteamine functionalized-gold nanoparticles for aflatoxin detection , 2010 .

[44]  Shouheng Sun,et al.  Interparticle interactions in coupled Au–Fe3O4 nanoparticles , 2009 .

[45]  R. Coker,et al.  The validation of a solid phase clean-up procedure for the analysis of aflatoxins in groundnut cake using HPLC , 1995 .

[46]  Yanbin Li,et al.  A self-assembled monolayer-based piezoelectric immunosensor for rapid detection of Escherichia coli O157:H7. , 2004, Biosensors & bioelectronics.

[47]  Juan Tang,et al.  Ultrasensitive electrochemical immunoassay of staphylococcal enterotoxin B in food using enzyme-nanosilica-doped carbon nanotubes for signal amplification. , 2010, Journal of agricultural and food chemistry.

[48]  D. B. Hibbert,et al.  Gold-coated magnetic nanoparticles as “dispersible electrodes” – Understanding their electrochemical performance , 2011 .

[49]  Elke Anklam,et al.  Acceptance of analytical methods for implementation of EU legislation with a focus on mycotoxins , 2002 .

[50]  Hong Jiang,et al.  Immune-biosensor for aflatoxin B1 based bio-electrocatalytic reaction on micro-comb electrode , 2006 .

[51]  Mihaela Puiu,et al.  Kinetic approach of aflatoxin B1-acetylcholinesterase interaction: a tool for developing surface plasmon resonance biosensors. , 2012, Analytical biochemistry.

[52]  J. Vidal,et al.  An electrochemical immunosensor for ochratoxin A determination in wines based on a monoclonal antibody and paramagnetic microbeads , 2012, Analytical and Bioanalytical Chemistry.

[53]  B. D. Malhotra,et al.  Carboxylated multiwalled carbon nanotubes based biosensor for aflatoxin detection , 2013 .

[54]  Uri Banin,et al.  Colloidal hybrid nanostructures: a new type of functional materials. , 2010, Angewandte Chemie.

[55]  G. Shen,et al.  Rapid detection of picloram in agricultural field samples using a disposable immunomembrane-based electrochemical sensor. , 2008, Environmental science & technology.

[56]  翁文桂,et al.  Advances in the Surface Engineering of Upconversion Nanocrystals , 2012 .

[57]  Jean-Louis Marty,et al.  Novel highly-performing immunosensor-based strategy for ochratoxin A detection in wine samples. , 2008, Biosensors & bioelectronics.

[58]  Yali Cui,et al.  Controllable synthesis and characterization of Fe3O4/Au composite nanoparticles , 2015 .

[59]  Jacques Ferlay,et al.  Estimating the world cancer burden: Globocan 2000 , 2001, International journal of cancer.

[60]  G. Palleschi,et al.  Detection of Aflatoxin B1 in Barley: Comparative Study of Immunosensor and HPLC , 2006 .

[61]  Xianxue Gan,et al.  Biomolecule-functionalized magnetic nanoparticles for flow-through quartz crystal microbalance immunoassay of aflatoxin B1 , 2009, Bioprocess and biosystems engineering.

[62]  N. Habibi,et al.  Fabrication and characterization of zinc oxide nanoparticle coated magnetic iron oxide: Effect of S-layers adsorption on surface of oxide , 2014 .

[63]  Joseph Owino,et al.  Electrochemical Immunosensor Based on Polythionine/Gold Nanoparticles for the Determination of Aflatoxin B1 , 2008, Sensors.

[64]  Xiulan Sun,et al.  Preparation of gold-labeled antibody probe and its use in immunochromatography assay for detection of aflatoxin B1. , 2005, International journal of food microbiology.

[65]  Yoshio Kobayashi,et al.  Direct Coating of Particles by a Liquid Phase Process , 2007 .

[66]  B. Kabak,et al.  Quantitation of aflatoxins in pistachios and groundnuts using HPLC-FLD method , 2014 .

[67]  Michael Pedersen,et al.  Targeting of peptide conjugated magnetic nanoparticles to urokinase plasminogen activator receptor (uPAR) expressing cells. , 2013, Nanoscale.

[68]  Jiayao Liao.  Construction of nanogold hollow balls with dendritic surface as immobilized affinity support for protein adsorption. , 2007, Colloids and surfaces. B, Biointerfaces.

[69]  J. Vidal,et al.  Use of polyclonal antibodies to ochratoxin A with a quartz–crystal microbalance for developing real-time mycotoxin piezoelectric immunosensors , 2009, Analytical and bioanalytical chemistry.

[70]  Saurabh Srivastava,et al.  Electrophoretically deposited reduced graphene oxide platform for food toxin detection. , 2013, Nanoscale.

[71]  Juan Tang,et al.  Conductive carbon nanoparticles-based electrochemical immunosensor with enhanced sensitivity for alpha-fetoprotein using irregular-shaped gold nanoparticles-labeled enzyme-linked antibodies as signal improvement. , 2010, Biosensors & bioelectronics.

[72]  Hans P. van Egmond,et al.  Regulations relating to mycotoxins in food , 2007 .

[73]  Yasar Gurbuz,et al.  Surface plasmon resonance based immunosensor for the detection of the cancer biomarker carcinoembryonic antigen. , 2011, Talanta.