Folding-Based Electrochemical Aptasensor for the Determination of β-Lactoglobulin on Poly-L-Lysine Modified Graphite Electrodes

Nowadays, food allergy is a very important health issue, causing adverse reactions of the immune system when exposed to different allergens present in food. Because of this, the development of point-of-use devices using miniaturized, user-friendly, and low-cost instrumentation has become of outstanding importance. According to this, electrochemical aptasensors have been demonstrated as useful tools to quantify a broad variety of targets. In this work, we develop a simple methodology for the determination of β-lactoglobulin (β-LG) in food samples using a folding-based electrochemical aptasensor built on poly-L-lysine modified graphite screen-printed electrodes (GSPEs) and an anti-β-lactoglobulin aptamer tagged with methylene blue (MB). This aptamer changes its conformation when the sample contains β-LG, and due to this, the spacing between MB and the electrode surface (and therefore the electron transfer efficiency) also changes. The response of this biosensor was linear for concentrations of β-LG within the range 0.1–10 ng·mL−1, with a limit of detection of 0.09 ng·mL−1. The biosensor was satisfactorily employed for the determination of spiked β-LG in real food samples.

[1]  Joana Costa,et al.  Detection and Quantification of Milk Ingredients as Hidden Allergens in Meat Products by a Novel Specific Real-Time PCR Method , 2019, Biomolecules.

[2]  Giovanna Marrazza,et al.  Recent advances of immunosensors for detecting food allergens , 2018, Current Opinion in Electrochemistry.

[3]  S. Taylor,et al.  Introduction to food allergy. , 2006 .

[4]  Alina Vasilescu,et al.  Electrochemical Affinity Biosensors Based on Disposable Screen-Printed Electrodes for Detection of Food Allergens , 2016, Sensors.

[5]  Georgina M. S. Ross,et al.  Consumer-friendly food allergen detection: moving towards smartphone-based immunoassays , 2018, Analytical and Bioanalytical Chemistry.

[6]  P. Erden,et al.  Disposable Amperometric Biosensor Based on Poly‐L‐lysine and Fe 3 O 4 NPs‐chitosan Composite for the Detection of Tyramine in Cheese , 2019, Electroanalysis.

[7]  Yunjia Yang,et al.  A rapid immobilized trypsin digestion combined with liquid chromatography – Tandem mass spectrometry for the detection of milk allergens in baked food , 2019, Food Control.

[8]  Steve L. Taylor,et al.  The Key Events Dose-Response Framework: A Foundation for Examining Variability in Elicitation Thresholds for Food Allergens , 2009, Critical reviews in food science and nutrition.

[9]  J. Qin,et al.  New polyacetylene-based chemosensory materials for the “turn-on” sensing of α-amino acids , 2009 .

[10]  H. Letheby XXIX.—On the production of a blue substance by the electrolysis of sulphate of aniline , 1862 .

[11]  Joana Costa,et al.  Cow's milk allergens: Screening gene markers for the detection of milk ingredients in complex meat products , 2020 .

[12]  Giovanna Marrazza,et al.  Amplified Electrochemical DNA Sensor Based on Polyaniline Film and Gold Nanoparticles , 2013 .

[13]  Weina Ma,et al.  Electrodeposited reduced graphene oxide incorporating polymerization of l-lysine on electrode surface and its application in simultaneous electrochemical determination of ascorbic acid, dopamine and uric acid. , 2017, Materials science & engineering. C, Materials for biological applications.

[14]  Scott H Sicherer,et al.  Clinical reviews in allergy and immunology , 2022 .

[15]  M. de Frutos,et al.  A new sample preparation method compatible with capillary electrophoresis and laser-induced fluorescence for improving detection of low levels of beta-lactoglobulin in infant foods. , 2009, Analytica chimica acta.

[16]  Joana Costa,et al.  Advances in Food Allergen Analysis , 2016 .

[17]  M. T. Fernández-Abedul,et al.  Methylene blue covalently attached to single stranded DNA as electroactive label for potential bioassays , 2013, Sensors and Actuators B: Chemical.

[18]  Joseph Wang,et al.  New label-free DNA recognition based on doping nucleic-acid probes within conducting polymer films , 1999 .

[19]  Yoon-Bo Shim,et al.  Conducting polymer-based electrochemical biosensors for neurotransmitters: A review. , 2018, Biosensors & bioelectronics.

[20]  Angelo Visconti,et al.  Immunochemical and DNA-based methods in food allergen analysis and quality assurance perspectives. , 2010 .

[21]  Steve L Taylor,et al.  Allergen immunoassays—considerations for use of naturally incurred standards , 2009, Analytical and bioanalytical chemistry.

[22]  Xiliang Luo,et al.  Zwitterionic peptide anchored to conducting polymer PEDOT for the development of antifouling and ultrasensitive electrochemical DNA sensor. , 2017, Biosensors & bioelectronics.

[23]  María Cruz Moreno-Bondi,et al.  Biosensing Based on Nanoparticles for Food Allergens Detection , 2018, Sensors.

[24]  L. Jacxsens,et al.  Analysis to support allergen risk management: Which way to go? , 2013, Journal of agricultural and food chemistry.

[25]  G. Niaura,et al.  Investigation of electrochemical polymerisation of L-lysine and application for immobilisation of functionalised graphene as platform for electrochemical sensing , 2019, Electrochimica Acta.

[26]  A. Heeger,et al.  Label-free electronic detection of thrombin in blood serum by using an aptamer-based sensor. , 2005, Angewandte Chemie.

[27]  T. Haahtela,et al.  Selection of recombinant IgE antibodies binding the beta-lactoglobulin allergen in a conformation-dependent manner. , 2009, Journal of immunological methods.

[28]  A. Sancho,et al.  Proteomic approaches for qualitative and quantitative characterisation of food allergens. , 2010, Regulatory toxicology and pharmacology : RTP.

[29]  R. Pieters,et al.  Cross-linking of β-lactoglobulin enhances allergic sensitization through changes in cellular uptake and processing. , 2014, Toxicological sciences : an official journal of the Society of Toxicology.

[30]  M. Samadpour,et al.  Development and Validation of a Lateral Flow Immunoassay Test Kit for Dual Detection of Casein and β-Lactoglobulin Residues. , 2016, Journal of food protection.

[31]  Kelley J. Rountree,et al.  A Practical Beginner’s Guide to Cyclic Voltammetry , 2017 .

[32]  Giovanna Marrazza,et al.  Electrochemical Nanocomposite Single-Use Sensor for Dopamine Detection , 2019, Sensors.

[33]  M. Ates A review study of (bio)sensor systems based on conducting polymers. , 2013, Materials science & engineering. C, Materials for biological applications.

[34]  Alexander D. MacKerell Influence of Magnesium Ions on Duplex DNA Structural, Dynamic, and Solvation Properties , 1997 .

[35]  Susana Campuzano,et al.  Electrochemical Affinity Biosensors in Food Safety , 2017 .

[36]  Tao Yang,et al.  A DNA electrochemical sensor with poly-l-lysine/single-walled carbon nanotubes films and its application for the highly sensitive EIS detection of PAT gene fragment and PCR amplification of NOS gene , 2008 .

[37]  E. Ferapontova,et al.  An RNA aptamer-based electrochemical biosensor for detection of theophylline in serum. , 2008, Journal of the American Chemical Society.

[38]  P. Kelly,et al.  Improving the sensory quality, shelf-life and functionality of milk , 2009 .

[39]  M. Ramezani,et al.  Detection of food-born allergens with aptamer-based biosensors , 2018 .

[40]  Patricia Schubert-Ullrich,et al.  Commercialized rapid immunoanalytical tests for determination of allergenic food proteins: an overview , 2009, Analytical and bioanalytical chemistry.

[41]  J. Soloducho,et al.  Conducting Polymers as Elements of Miniature Biocompatible Sensor , 2018, Green Electronics.

[42]  J. Marty,et al.  Sensitive analytical performance of folding based biosensor using methylene blue tagged aptamers. , 2016, Talanta.

[43]  A. Sheikh,et al.  Prevalence of common food allergies in Europe: a systematic review and meta‐analysis , 2014, Allergy.

[44]  A. Urbani,et al.  Precision medicine in cow's milk allergy: proteomics perspectives from allergens to patients. , 2018, Journal of proteomics.

[45]  Giovanna Marrazza,et al.  Oligonucleotide-modified screen-printed gold electrodes for enzyme-amplified sensing of nucleic acids. , 2004, Biosensors & bioelectronics.

[46]  Anshu Yang,et al.  Highly Sensitive Detection of Bovine β-Lactoglobulin with Wide Linear Dynamic Range Based on Platinum Nanoparticles Probe. , 2018, Journal of agricultural and food chemistry.

[47]  Filiz Kuralay,et al.  Poly‐L‐lysine Coated Surfaces for Ultrasensitive Nucleic Acid Detection , 2018, Electroanalysis.

[48]  H. Sakai,et al.  Polylysine Produced by Streptomyces , 1977 .

[49]  C. Bala,et al.  Electrochemical biosensors for fast detection of food contaminants trends and perspective , 2016 .

[50]  Joana Costa,et al.  Bovine Milk Allergens: A Comprehensive Review. , 2018, Comprehensive reviews in food science and food safety.

[51]  Xuan Weng,et al.  Nano-biosensor platforms for detecting food allergens – New trends , 2018 .

[52]  Ryan J. White,et al.  Enhancing the analytical performance of electrochemical RNA aptamer-based sensors for sensitive detection of aminoglycoside antibiotics. , 2014, Analytical chemistry.

[53]  Steve L Taylor,et al.  Food , drug , insect sting allergy , and anaphylaxis Allergen reference doses for precautionary labeling ( VITAL 2 . 0 ) : Clinical implications , 2022 .

[54]  H. Sampson,et al.  Treatments for food allergy: how close are we? , 2012, Immunologic research.

[55]  Georgina M. S. Ross,et al.  A Critical Comparison between Flow-through and Lateral Flow Immunoassay Formats for Visual and Smartphone-Based Multiplex Allergen Detection , 2019, Biosensors.

[56]  Darren R. Flower,et al.  Bovine β-lactoglobulin at 1.8 Å resolution — still an enigmatic lipocalin , 1997 .

[57]  B. McDuffie,et al.  Diffusion coefficients of ferri- and ferrocyanide ions in aqueous media, using twin-electrode thin-layer electrochemistry , 1970 .

[58]  José M Pingarrón,et al.  Electrochemical magnetoimmunosensing platform for determination of the milk allergen β-lactoglobulin. , 2015, Talanta.

[59]  G. Marrazza,et al.  Beta‐lactoglobulin Electrochemical Detection Based with an Innovative Platform Based on Composite Polymer , 2020, Electroanalysis.

[60]  Chunhai Fan,et al.  Aptamer-based biosensors , 2008 .

[61]  Wei Liu,et al.  Sensitive electrochemical aptasensor by coupling "signal-on'' and "signal-off'' strategies. , 2013, Analytical chemistry.

[62]  Jian Ji,et al.  Development of a liquid chromatography-tandem mass spectrometry method for simultaneous detection of the main milk allergens , 2017 .

[63]  A. Azadbakht,et al.  Aptamer-Based Approach as Potential Tools for Construction the Electrochemical Aptasensor , 2018, Journal of Inorganic and Organometallic Polymers and Materials.