Optimization and characterization of a biosensor assembly for detection of <Emphasis Type="Italic">Salmonella</Emphasis> Typhimurium |
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Authors: | A M A Melo D L Alexandre M R F Oliveira R F Furtado M F Borges P R V Ribeiro A Biswas H N Cheng C R Alves E A T Figueiredo |
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Institution: | 1.Department of Science and Food Technology,Federal University of Ceará,Fortaleza,Brazil;2.Department of Chemistry,Fortaleza,Brazil;3.Embrapa Tropical Agroindustry,Fortaleza,Brazil;4.USDA Agricultural Research Service,National Center for Agricultural Utilization Research,Peoria,USA;5.USDA Agricultural Research Service, Southern Regional Research Center,New Orleans,USA |
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Abstract: | The performance of biosensors depends directly on the strategies adopted during their development. In this paper, a fast and sensitive biosensor for Salmonella Typhimurium detection was assembled by using optimization studies in separate stages. The pre-treatment assays, biomolecular immobilization (primary antibody and protein A concentrations), and analytical response (hydroquinone and hydrogen peroxide concentrations) were optimized via voltammetric methods. In the biosensor assembly, a gold surface was modified via the self-assembled monolayer technique (SAM) using cysteamine thiol and protein A for immobilization of anti-Salmonella antibody. The analytical response of the biosensor was obtained through the use of a secondary antibody labeled with a peroxidase enzyme, and the signal was evaluated by applying the chronoamperometry technique. The biosensor was characterized by infrared spectroscopy and cyclic voltammetry. Optimization of protein A and primary antibody concentrations enabled higher analytical signals of 7.5 and 75 mg mL?1, respectively, to be achieved. The hydroquinone and H2O2 concentrations selected were 3 and 300 mM, respectively. The biosensor developed attained a very low detection limit of 10 CFU mL?1 and a fast response with a final detection time of 125 min. These results indicate that this biosensor is very promising for the food safety and emergency response applications. |
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