Magnetic Bead-Based Colorimetric Immunoassay for Aflatoxin B1 Using Gold Nanoparticles

A competitive colorimetric immunoassay for the detection of aflatoxin B1 (AFB) has been established using biofunctionalized magnetic beads (MBs) and gold nanoparticles (GNPs). Aflatoxin B1-bovine serum albumin conjugates (AFB-BSA) modified MBs were employed as capture probe, which could specifically bind with GNP-labeled anti-AFB antibodies through immunoreaction, while such specific binding was competitively inhibited by the addition of AFB. After magnetic separation, the supernatant solution containing unbound GNPs was directly tested by UV-Vis spectroscopy. The absorption intensity was directly proportional to the AFB concentration. The influence of GNP size, incubation time and pH was investigated in detail. After optimization, the developed method could detect AFB in a linear range from 20 to 800 ng/L, with the limit of detection at 12 ng/L. The recoveries for spiked maize samples ranged from 92.8% to 122.0%. The proposed immunoassay provides a promising approach for simple, rapid, specific and cost-effective detection of toxins in the field of food safety.

[1]  Guohua Zhou,et al.  Simple, rapid, homogeneous oligonucleotides colorimetric detection based on non-aggregated gold nanoparticles. , 2012, Chemical communications.

[2]  Yu Zhang,et al.  Selective collection and detection of leukemia cells on a magnet-quartz crystal microbalance system using aptamer-conjugated magnetic beads. , 2010, Biosensors & bioelectronics.

[3]  Andrew Wang,et al.  Immunochromatographic Assay for Ultrasensitive Detection of Aflatoxin B1 in Maize by Highly Luminescent Quantum Dot Beads , 2014, ACS applied materials & interfaces.

[4]  Jian Ling,et al.  Magnetic particle-based sandwich sensor with DNA-modified carbon nanotubes as recognition elements for detection of DNA hybridization. , 2008, Analytical chemistry.

[5]  Laura Anfossi,et al.  Lateral Flow Immunoassays for Aflatoxins B and G and for Aflatoxin M1 , 2013 .

[6]  Tapas Sen,et al.  Multifunctional magnetite and silica–magnetite nanoparticles: Synthesis, surface activation and applications in life sciences , 2005 .

[7]  Reinhard Niessner,et al.  Comparison of hybridoma screening methods for the efficient detection of high-affinity hapten-specific monoclonal antibodies. , 2008, Journal of immunological methods.

[8]  Peng Wang,et al.  Fluorescent artificial enzyme-linked immunoassay system based on Pd/C nanocatalyst and fluorescent chemodosimeter. , 2013, Analytical chemistry.

[9]  Jianzhong Lu,et al.  Magnetic bead-based chemiluminescent metal immunoassay with a colloidal gold label. , 2005, Analytical chemistry.

[10]  Chad A Mirkin,et al.  Homogeneous detection of nucleic acids based upon the light scattering properties of silver-coated nanoparticle probes. , 2007, Analytical chemistry.

[11]  Guonan Chen,et al.  Magnetic bead-based reverse colorimetric immunoassay strategy for sensing biomolecules. , 2013, Analytical chemistry.

[12]  Chad A Mirkin,et al.  The bio-barcode assay for the detection of protein and nucleic acid targets using DTT-induced ligand exchange , 2006, Nature Protocols.

[13]  Sang Jun Sim,et al.  Homogenous growth of gold nanocrystals for quantification of PSA protein biomarker. , 2009, Biosensors & bioelectronics.

[14]  Bing Xu,et al.  Nitrilotriacetic acid-modified magnetic nanoparticles as a general agent to bind histidine-tagged proteins. , 2004, Journal of the American Chemical Society.

[15]  Xiaoli Zhu,et al.  Magnetic nanoparticles applied in electrochemical detection of controllable DNA hybridization. , 2006, Analytical chemistry.

[16]  Yunyan Huang,et al.  Highly sensitive protein detection using enzyme-labeled gold nanoparticle probes. , 2010, The Analyst.

[17]  Xiaoru Wang,et al.  Amplified detection of protein cancer biomarkers using DNAzyme functionalized nanoprobes. , 2009, Chemical communications.

[18]  I. Kennedy,et al.  A rapid aflatoxin B1 ELISA: development and validation with reduced matrix effects for peanuts, corn, pistachio, and Soybeans. , 2004, Journal of agricultural and food chemistry.

[19]  C. Bala,et al.  Sensitive Aflatoxin B1 Determination Using a Magnetic Particles-Based Enzyme-Linked Immunosorbent Assay , 2008, Sensors.

[20]  I. Hussain,et al.  Aflatoxin Measurement and Analysis , 2011 .

[21]  Bahruddin Saad,et al.  Determination of aflatoxins in animal feeds by HPLC with multifunctional column clean-up , 2010 .

[22]  D. Fernig,et al.  Determination of size and concentration of gold nanoparticles from UV-vis spectra. , 2007, Analytical chemistry.

[23]  G. Frens.  Controlled Nucleation for the Regulation of the Particle Size in Monodisperse Gold Suspensions , 1973 .

[24]  Xin Li,et al.  Molecular characterization of monoclonal antibodies against aflatoxins: a possible explanation for the highest sensitivity. , 2012, Analytical chemistry.

[25]  Mei Liu,et al.  Homogeneous fluorescence-based immunoassay via inner filter effect of gold nanoparticles on fluorescence of CdTe quantum dots. , 2012, The Analyst.

[26]  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.

[27]  Reinhard Niessner,et al.  Review: bioanalytical applications of biomolecule-functionalized nanometer-sized doped silica particles. , 2009, Analytica chimica acta.

[28]  Yibin Ying,et al.  Comparison of monomeric and polymeric horseradish peroxidase as labels in competitive ELISA for small molecule detection , 2013, Microchimica Acta.

[29]  Patrick Couvreur,et al.  Magnetic nanoparticles: design and characterization, toxicity and biocompatibility, pharmaceutical and biomedical applications. , 2012, Chemical reviews.

[30]  Taeghwan Hyeon,et al.  Multifunctional nanostructured materials for multimodal imaging, and simultaneous imaging and therapy. , 2009, Chemical Society reviews.

[31]  X Chris Le,et al.  Aptamer-linked assay for thrombin using gold nanoparticle amplification and inductively coupled plasma-mass spectrometry detection. , 2009, Analytical chemistry.

[32]  Jianzhong Lu,et al.  Hydroxylamine-amplified gold nanoparticles for the homogeneous detection of sequence-specific DNA. , 2010, The Analyst.

[33]  Hui Li,et al.  A non-aggregation colorimetric assay for thrombin based on catalytic properties of silver nanoparticles. , 2014, Analytica chimica acta.

[34]  Jicun Ren,et al.  Sandwich immunoassay for alpha-fetoprotein in human sera using gold nanoparticle and magnetic bead labels along with resonance Rayleigh scattering readout , 2013, Microchimica Acta.

[35]  Jie Li,et al.  Nanoparticle-catalyzed reductive bleaching for fabricating turn-off and enzyme-free amplified colorimetric bioassays. , 2014, Biosensors & bioelectronics.

[36]  Sha Zhang,et al.  Detection of aflatoxin M1 in milk by dynamic light scattering coupled with superparamagnetic beads and gold nanoprobes. , 2013, Journal of agricultural and food chemistry.

[37]  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.

[38]  Qun Huo,et al.  Gold nanoparticle-enabled biological and chemical detection and analysis. , 2012, Chemical Society reviews.

[39]  Chad A Mirkin,et al.  Nanostructures in biodiagnostics. , 2005, Chemical reviews.

[40]  Jianzhong Lu,et al.  Sequential determination of two proteins by temperature-triggered homogeneous chemiluminescent immunoassay. , 2006, Analytical chemistry.

[41]  Jing-Juan Xu,et al.  Enzyme-free colorimetric bioassay based on gold nanoparticle-catalyzed dye decolorization. , 2013, The Analyst.

[42]  Jiashu Sun,et al.  Point-of-care biochemical assays using gold nanoparticle-implemented microfluidics. , 2014, Chemical Society reviews.

[43]  Huanghao Yang,et al.  Magnetic bead-based enzyme-chromogenic substrate system for ultrasensitive colorimetric immunoassay accompanying cascade reaction for enzymatic formation of squaric acid-iron(III) chelate. , 2014, Analytical chemistry.

[44]  Juan Tang,et al.  Magneto-controlled graphene immunosensing platform for simultaneous multiplexed electrochemical immunoassay using distinguishable signal tags. , 2011, Analytical chemistry.

[45]  R. Niessner,et al.  Multifunctional magnetic bead-based electrochemical immunoassay for the detection of aflatoxin B1 in food. , 2009, The Analyst.

[46]  Igor L. Medintz,et al.  Functionalizing nanoparticles with biological molecules: developing chemistries that facilitate nanotechnology. , 2013, Chemical reviews.

[47]  Ronen Polsky,et al.  Magnetically-induced solid-state electrochemical detection of DNA hybridization. , 2002, Journal of the American Chemical Society.

[48]  Zhiqiang Gao,et al.  Nanoparticles in biomolecular detection , 2006 .

[49]  A. Zherdev,et al.  Immunochromatographic methods in food analysis , 2014 .

[50]  Liesbet Lagae,et al.  A simple double-bead sandwich assay for protein detection in serum using UV-vis spectroscopy. , 2011, Talanta.

[51]  Abraham Ulman,et al.  Activity of Candida rugosa lipase immobilized on gamma-Fe2O3 magnetic nanoparticles. , 2003, Journal of the American Chemical Society.

[52]  Sai Bi,et al.  Gold nanolabels for new enhanced chemiluminescence immunoassay of alpha-fetoprotein based on magnetic beads. , 2009, Chemistry.

[53]  Sang Bok Lee,et al.  Magnetic nanotubes for magnetic-field-assisted bioseparation, biointeraction, and drug delivery. , 2005, Journal of the American Chemical Society.

[54]  R. Niessner,et al.  Magnetic bead-based fluorescence immunoassay for aflatoxin B1 in food using biofunctionalized rhodamine B-doped silica nanoparticles. , 2010, The Analyst.

[55]  Chiara Cavaliere,et al.  Determination of aflatoxins in hazelnuts by various sample preparation methods and liquid chromatography-tandem mass spectrometry. , 2008, Journal of chromatography. A.

[56]  Hye-Weon Yu,et al.  Bead-based competitive fluorescence immunoassay for sensitive and rapid diagnosis of cyanotoxin risk in drinking water. , 2011, Environmental science & technology.

[57]  Lin He,et al.  Nanoparticles for bioanalysis. , 2003, Current opinion in chemical biology.