Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 545405
Title Functional fluorescence assay of botulinum neurotoxin A in complex matrices using magnetic beads
Author(s) Klisara, Nevena; Peters, Jeroen; Haasnoot, Willem; Nielen, Michel W.F.; Palaniappan, Alagappan; Liedberg, Bo
Source Sensors and Actuators B: Chemical 281 (2019). - ISSN 0925-4005 - p. 912 - 919.
DOI https://doi.org/10.1016/j.snb.2018.10.100
Department(s) BU Authenticity & Bioassays
VLAG
Monsteradministratie & Coördinatie
Organic Chemistry
Publication type Refereed Article in a scientific journal
Publication year 2019
Keyword(s) Botulinum neurotoxin A - Fluorescence detection - Peptide - Superparamagnetic beads
Abstract

The extremely toxic botulinum neurotoxin poses a threat for health and food safety, requiring rapid and easy-to-use detection platforms. To meet these requirements, we have explored a novel functional assay format for detection of botulinum neurotoxin serotype A light chain (BoNT/A LC) in complex matrices. The proposed assay utilizes a synthetic peptide designed to mimic the SNAP-25 protein (synaptosomal-associated protein 25) as substrate bound to a superparamagnetic bead and a fluorescent dye. Cleavage of the peptide by BoNT/A LC yields a reduction in fluorescence signal revealing the presence of the BoNT/A LC in the sample matrices tested. The superparamagnetic beads enable efficient separation of the cleaved peptides from food matrices, thereby improving the reliability of responses. Herein, we demonstrate a protocol offering an assay time of 6 h and a LOD of 0.5 nM (25 ng/ml). The proposed protocol is validated using carrot juice and diluted milk pending further improvements in sensitivity and overall assay time. Robustness, cost-effectiveness, low sample volume requirements in conjunction with the possibility of multiplexing for other proteolytic enzymes makes the proposed protocol competitive in comparison with conventional BoNT assays reported elsewhere.

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