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Abstract

In the last decades, the community has been facing the problems arising from the transfer of potentially harmful substances to the environment, altering the ecosystem, and to the human, causing pathological symptoms, and sometimes death. Some of the most diffuse neurotoxic chemical compounds are organophosphates, including pesticides and nervine gas. These phosphorus-based compounds are the most common and powerful acetylcholinesterase inhibitors, being designed to bind with high affinity to the active site of this enzyme. It was worked to use the esterase 2 from Alicyclobacillus acidocaldarius (EST2) as a biosensor for the detection of specific organophosphate pesticides. With the recent demonstration of the very high affinity of EST2 toward paraoxon, a more complete analysis on the detection methods in water was carried out. The inhibitory effects of a wide range of other pesticides on EST2 were investigated, showing a better selectivity with respect to nonspecific reaction of acethylcholinesterases. The applied methodology allowed one to detect 2.75 10-3 ppm of neurotoxic agent, comparable to the efficiency of other acethylcholinesterase-based biosensors and allowed the development of a raw biosensor, based on EST2 immobilization on a nitrocellulose membrane, for paraoxon detection. Also, using a robotic automatized assay on a MICROLAB® STAR Liquid Handling Workstations equipped by a robotic arm and a VICTORTM X3 Multilabel Plate Reader, it was developed a method for the identification of almost 12 pesticides in a single 96 well plate and their specificity in order to realize a new type of biosensor based on the LAB ON CHIP concept.

Biography

Ferdinando Febbraio has completed his PhD in Biochemistry and Molecular Biology in 2001 at University of Naples. He is Researcher at Institute of Protein Biochemistry of National Research Council. He has published more than 40 papers in international journals.