Detection of aflatoxin B1 by aptamer-based biosensor using PAMAM dendrimers as immobilization platform |
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| Affiliation: | 1. Faculty of Mathematics, Physics and Informatics, Comenius University, Mlynska Dolina F1, 842 48 Bratislava, Slovakia;2. Università di Roma TorVergata, Dipartimento di Scienze e Tecnologie Chimiche, Via della Ricerca Scientifica 1, 00133 Rome, Italy;3. ENEA, Italian National Agency for New Technologies, Energy and the Environment, Centro Ricerche Casaccia, Via Anguillarese 301, 00123 Rome, Italy;4. Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, Moyzesova 61, 900 28 Ivanka pri Dunaji, Slovakia;1. Ministry of Agriculture Laboratory of Quality & Safety Risk Assessment for Dairy Products (Beijing), Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, PR China;2. College of Animal Science and Technology, Xinjiang Agricultural University, Urumchi 830000, PR China;3. Ministry of Agriculture—Milk and Dairy Product Inspection Center (Beijing), Beijing 100193, PR China;4. State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, PR China;1. BAE Laboratory, Université de Perpignan Via Domitia, 52 Avenue Paul Alduy, Perpignan 66860, France;2. Biosensor Lab, Department of Chemistry, BITS, Pilani, K. K. Birla Goa Campus, Zuarinagar, Goa 403726, India;3. Interdisciplinary Research Centre in Biomedical Materials (IRCBM), COMSATS Institute of Information Technology (CIIT), Lahore 54000, Pakistan;4. Biocatalysis and Industrial Enzymes Group, Laboratory of Microbial Ecology and Technology, Carthage University, National Institute of Applied Sciences and Technology, BP 676, 1080 Tunis, Tunisia;1. Pharmaceutical Research Center, Mashhad University of Medical Sciences, Mashhad, Iran;2. Department of Medicinal Chemistry, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran;3. Nanotechnology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran;4. Research Institute of Sciences and New Technology, Mashhad, Iran;5. Cardiovascular Research Center, Ghaem hospital, Mashhad University of Medical Sciences, Mashhad, Iran;6. Department of Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran;7. Targeted Drug Delivery Research Center, Mashhad University of Medical Sciences, Mashhad, Iran;8. Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran;1. Department of Chemistry, School of Physics and Chemistry, Gwangju Institute of Science and Technology (GIST), 261 Cheomdan-gwagiro, Gwangju 500-712, Republic of Korea;2. Advanced Photonics Research Institute, Gwangju Institute of Science and Technology (GIST), 261 Cheomdan-gwagiro, Gwangju 500-712, Republic of Korea;3. Food Analysis Research Team, Industry Service Research Center, World Institute of Kimchi an Annex of Korea Food Research Institute, 86, Kimchi-ro, Nam-gu, Gwangju, Republic of Korea;1. Department of Chemistry, School of Physics and Chemistry, Gwangju Institute of Science and Technology (GIST), Gwangju 500-712, Republic of Korea;2. Advanced Photonics Research Institute, Gwangju Institute of Science and Technology (GIST), Gwangju 500-712, Republic of Korea;1. School of Biotechnology and Food Engineering, Anhui Provincial Key Lab of Functional Materials and Devices, Hefei University of Technology, Hefei 23009, China;2. Jiangsu Entry-Exit Inspection and Quarantine Bureau, Nanjing 200002, China;3. Department of Chemistry and Biochemistry, North Dakota State University, Fargo, ND 58102, USA |
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| Abstract: | We report an aptamer-based biosensor for detection of aflatoxin B1 (AFB1), a mycotoxin identified as contaminant in food. The sensor is assembled in a multilayer framework that utilizes cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) for acquiring the signal response by means of redox indicators: K[Fe(CN)6]−3/−4. Poly (amidoamine) dendrimers of fourth generation (PAMAM G4) immobilized on gold electrode covered by cystamine, were employed for attachment of single stranded amino-modified DNA aptamers specific to AFB1. The cystamine-dendrimers (Cys-PAMAM) layers were compared with other immobilization platforms such as cystamine (Cys), 11-mercaptoundecanoic acid (MUA) and 11-mercaptoundecanoic acid-dendrimers (MUA-PAMAM), being the first approach the most appropriate for producing sensitive and reproducible signal in the range of concentrations 0.1–10 nM AFB1. The sensor was validated in certified contaminated peanuts extract as well as in spiked samples of peanuts-corn snacks and the sensing response was evaluated and compared in terms of the matrix effect. The aptamer specificity was analyzed by testing the sensor in other mycotoxins such as aflatoxin B2 (AFB2) and ochratoxin A (OTA). The limit of detection achieved by this sensor was LOD = 0.40 ± 0.03 nM, it was regenerable in 0.2 M glycine-HCl and it did not lose its stability up to 60 h storing at 4 °C. Atomic Force Microscopy (AFM) studies were also performed for illustrating individual steps of biosensor assembly. |
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| Keywords: | Aptamers PAMAM dendrimers Biosensor Electrochemistry Atomic force microscopy Contaminated peanuts |
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