Title:
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Conjugation of genetically-engineered protein phosphatases to magnetic particles for okadaic acid detection
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Author:
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Garibo, Diana; Devic, Eric; Marty, Jean-Louis; Diogène Fadini, Jorge; Unzueta, Idoia; Blázquez, María; Campàs i Homs, Mònica
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Other authors:
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IRTA. Recerca i Tecnologia Agroalimentàries |
Abstract:
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This work presents the functional characterisation of a protein phosphatase 2A (PP2A) catalytic subunit obtained by genetic engineering and its conjugation to magnetic particles (MPs) via metal coordination chemistry for the subsequent development of assays for diarrheic lipophilic marine toxins. Colorimetric assays with free enzyme have allowed the determination of the best enzyme activity stabiliser, which is glycerol at 10%. They have also demonstrated that the recombinant enzyme can be as sensitive towards okadaic acid (OA) (LOD=2.3μg/L) and dinophysistoxin-1 (DTX-1) (LOD=15.2μg/L) as a commercial PP2A and, moreover, it has a higher operational stability, which makes possible to perform the protein phosphatase inhibition assay (PPIA) with a lower enzyme amount. Once conjugated to MPs, the PP2A catalytic subunit still retains its enzyme activity and it can also be inhibited by OA (LOD=30.1μg/L). |
Publication date:
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2012-01 |
Subject(s):
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Toxines marines |
Rights:
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L'accés als continguts d'aquest document queda condicionat a l'acceptació de les condicions d'ús establertes per la següent llicència Creative Commons: http://creativecommons.org/licenses/by-nc-nd/3.0/es/ |
Pages:
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29 p. |
Document type:
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Article Article - Accepted version |
Published by:
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Journal of Biotechnology
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