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Open AccessArticle

Fluorescence Spectroscopy Approaches for the Development of a Real-Time Organophosphate Detection System Using an Enzymatic Sensor

The Institute of Protein Biochemistry (IBP) of the Italian National Research Council (CNR), Via P. Castellino 111, 80131 Naples, Italy
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Sensors 2015, 15(2), 3932-3951; https://doi.org/10.3390/s150203932
Received: 11 December 2014 / Accepted: 2 February 2015 / Published: 9 February 2015
(This article belongs to the Special Issue State-of-the-Art Sensors Technology in Italy 2014)
Organophosphates are organic substances that contain a phosphoryl or a thiophosphoryl bond. They are mainly used around the world as pesticides, but can also be used as chemical warfare agents. Their detection is normally entrusted to techniques like GC- and LC-MS that, although sensitive, do not allow their identification on site and in real time. We have approached their identification by exploiting the high-affinity binding of these compounds with the esterase 2 from Alicyclobacillus acidocaldarius. Using an in silico analysis to evaluate the binding affinities of the enzyme with organophosphate inhibitors, like paraoxon, and other organophosphate compounds, like parathion, chlorpyriphos, and other organophosphate thio-derivatives, we have designed fluorescence spectroscopy experiments to study the quenching of the tryptophan residues after esterase 2 binding with the organophosphate pesticides. The changes in the fluorescence signals permitted an immediate and quantitative identification of these compounds from nano- to picomolar concentrations. A fluorescence based polarity-sensitive probe (ANS) was also employed as a means to understand the extent of the interactions involved, as well as to explore other ways to detect organophosphate pesticides. Finally, we designed a framework for the development of a biosensor that exploits fluorescence technology in combination with a sensitive and very stable bio-receptor. View Full-Text
Keywords: biosensor; organophosphates; fluorescence spectroscopy; thermostable esterase 2; nerve agents; pesticides biosensor; organophosphates; fluorescence spectroscopy; thermostable esterase 2; nerve agents; pesticides
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Carullo, P.; Cetrangolo, G.P.; Mandrich, L.; Manco, G.; Febbraio, F. Fluorescence Spectroscopy Approaches for the Development of a Real-Time Organophosphate Detection System Using an Enzymatic Sensor. Sensors 2015, 15, 3932-3951.

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