Functional fluorescence assay of botulinum neurotoxin A in complex matrices using magnetic beads
| Autor: | Bo Liedberg, Nevena Klisara, Willem Haasnoot, Michel W. F. Nielen, Alagappan Palaniappan, Jeroen Peters |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Monsteradministratie & Coördinatie
Fluorescence assay Peptide 02 engineering and technology Superparamagnetic beads 010402 general chemistry 01 natural sciences Fluorescence detection BU Authenticity & Bioassays Materials Chemistry Electrical and Electronic Engineering Instrumentation VLAG chemistry.chemical_classification Botulinum neurotoxin A Complex matrix Chromatography Chemistry Organic Chemistry Metals and Alloys Proteolytic enzymes Substrate (chemistry) 021001 nanoscience & nanotechnology Condensed Matter Physics Organische Chemie Fluorescence Botulinum neurotoxin 0104 chemical sciences Surfaces Coatings and Films Electronic Optical and Magnetic Materials BU Authenticiteit & Bioassays 0210 nano-technology |
| Zdroj: | Sensors and Actuators, B: Chemical 281 (2019) Sensors and Actuators, B: Chemical, 281, 912-919 |
| ISSN: | 0925-4005 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect