Use of polyclonal antibodies to ochratoxin A with a quartz–crystal microbalance for developing real-time mycotoxin piezoelectric immunosensors

A piezoelectric immunosensor was tested for ochratoxin A (OTA) mycotoxin detection through the immobilization of OTA–bovine serum albumin (OTA–BSA) conjugate on gold-coated quartz crystals (AT-cut/5 MHz). Immunoassays were performed in a flow-injection system through frequency decreases in a quartz–crystal microbalance (QCM) because of a mass increasing during immunoreaction with anti-OTA antibodies. Three immobilization procedures for OTA–BSA (direct adsorption and covalent attachment to two alkane thiol self-assembled monolayers) were characterized with QCM in real time. Covalent attachment of the OTA–BSA conjugates through gold nanoparticles was also tested for amplifying the signal. Binding of the excess of antibodies to the immobilized OTA in an indirect competitive analysis decreased linearly the resonant frequency in the range of the OTA concentration from 10 to 128 ng/mL, with a detection limit of 8 ng/mL (signal/noise ratio of 3). A pepsin 2 mg/mL (pH = 2.1) solution was used to release antigen–antibody complexes, regenerating the biorecognition surface.

[1]  Jean-Louis Marty,et al.  Label-free impedimetric immunosensor for sensitive detection of ochratoxin A. , 2009, Biosensors & bioelectronics.

[2]  J. Hayes,et al.  Reduction in antioxidant defenses may contribute to ochratoxin A toxicity and carcinogenicity. , 2006, Toxicological sciences : an official journal of the Society of Toxicology.

[3]  C. Steinem,et al.  Piezoelectric Mass-Sensing Devices as Biosensors-An Alternative to Optical Biosensors? , 2000, Angewandte Chemie.

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

[5]  M. Ward,et al.  The Role of Longitudinal Waves in Quartz Crystal Microbalance Applications in Liquids , 1995 .

[6]  Biao Huang,et al.  Determination of ochratoxin A by polyclonal antibodies based sensitive time-resolved fluoroimmunoassay , 2006, Archives of Toxicology.

[7]  F. Ricci,et al.  A review on novel developments and applications of immunosensors in food analysis. , 2007, Analytica chimica acta.

[8]  Giuseppe Palleschi,et al.  Monoclonal antibody based electrochemical immunosensor for the determination of ochratoxin A in wheat. , 2006, Talanta.

[9]  B. Conway,et al.  Dependence of the reliability of electrochemical quartz-crystal nanobalance mass responses on the calibration constant, Cf: analysis of three procedures for its determination , 2003 .

[10]  M. Agut,et al.  Ultra-performance liquid chromatography/tandem mass spectrometry for the simultaneous analysis of aflatoxins B1, G1, B2, G2 and ochratoxin A in beer. , 2006, Rapid communications in mass spectrometry : RCM.

[11]  M. Ward,et al.  Self-assembled thiol monolayers with carboxylic acid functionality: measuring pH-dependent phase transitions with the quartz crystal microbalance , 1992 .

[12]  R. K. Mendes,et al.  Effects of different self-assembled monolayers on enzyme immobilization procedures in peroxidase-based biosensor development , 2008 .

[13]  Debopam Acharya,et al.  Simultaneous enzyme immunoassay for the screening of aflatoxin B1 and ochratoxin A in chili samples. , 2007, Analytica chimica acta.

[14]  Zhongming Zheng,et al.  Validation of an ELISA test kit for the detection of ochratoxin A in several food commodities by comparison with HPLC , 2005, Mycopathologia.

[15]  Loïc J Blum,et al.  State of the art and recent advances in immunoanalytical systems. , 2006, Biosensors & bioelectronics.

[16]  J. Anzai,et al.  Quartz-crystal microbalance and cyclic voltammetric studies of the adsorption behaviour of serum albumin on self-assembled thiol monolayers possessing different hydrophobicity and polarity , 1996 .

[17]  Francesco Palmisano,et al.  Determination of Ochratoxin A in wine at sub ng/mL levels by solid-phase microextraction coupled to liquid chromatography with fluorescence detection. , 2006, Journal of chromatography. A.

[18]  M. Váradi,et al.  Development of immunosensor based on OWLS technique for determining Aflatoxin B1 and Ochratoxin A. , 2007, Biosensors & bioelectronics.

[19]  S. Saeger,et al.  Novel gel-based rapid test for non-instrumental detection of ochratoxin A in beer , 2008, Analytical and bioanalytical chemistry.

[20]  F. Yu,et al.  Development of a sensitive enzyme-linked immunosorbent assay for the determination of ochratoxin A. , 2005, Journal of agricultural and food chemistry.

[21]  Hua Ai,et al.  Amplification of the antigen-antibody interaction from quartz crystal microbalance immunosensors via back-filling immobilization of nanogold on biorecognition surface. , 2006, Journal of immunological methods.

[22]  R. Manderville,et al.  Ochratoxin A: An overview on toxicity and carcinogenicity in animals and humans. , 2007, Molecular nutrition & food research.

[23]  K. Marx,et al.  Quartz crystal microbalance: a useful tool for studying thin polymer films and complex biomolecular systems at the solution-surface interface. , 2003, Biomacromolecules.

[24]  J. M. Soriano,et al.  Factors Affecting the Presence of Ochratoxin A in Wines , 2006, Critical reviews in food science and nutrition.

[25]  J. C. Hoogvliet,et al.  Electrochemical pretreatment of polycrystalline gold electrodes to produce a reproducible surface roughness for self-assembly: a study in phosphate buffer pH 7.4 , 2000, Analytical chemistry.

[26]  A. Hocking,et al.  Quantitative analysis of ochratoxin A in wine and beer using solid phase extraction and high performance liquid chromatography–fluorescence detection , 2006, Food additives and contaminants.

[27]  N. Miura,et al.  Recent advancements in surface plasmon resonance immunosensors for detection of small molecules of biomedical, food and environmental interest , 2007 .

[28]  Hiep Ly,et al.  Stability and Self-Exchange in Alkanethiol Monolayers , 1995 .

[29]  James L. Baker,et al.  Ochratoxins: A global perspective , 2006, Mycopathologia.

[30]  Linda Monaci,et al.  Determination of ochratoxin A in foods: state-of-the-art and analytical challenges , 2004, Analytical and bioanalytical chemistry.

[31]  S. Chung,et al.  Determination of ochratoxin A at parts-per-trillion levels in cereal products by immunoaffinity column cleanup and high-performance liquid chromatography/mass spectrometry. , 2007, Journal of AOAC International.

[32]  Wen‐Chi Tsai,et al.  QCM‐Based Immunosensor for the Determination of Ochratoxin A , 2007 .