title: Nanopore sensing of botulinum toxin type B by discriminating an enzymatically cleaved Peptide from a synaptic protein synaptobrevin 2 derivative.
abstract: Botulinum neurotoxins (BoNTs) are the most lethal toxin known to human. Biodefense requires early and rapid detection of BoNTs. Traditionally, BoNTs can be detected by looking for signs of botulism in mice that receive an injection of human material, serum or stool. While the living animal assay remains the most sensitive approach, it is costly, slow and associated with legal and ethical constrains. Various biochemical, optical and mechanical methods have been developed for BoNTs detection with improved speed, but with lesser sensitivity. Here, we report a novel nanopore-based BoNT type B (BoNT-B) sensor that monitors the toxin's enzymatic activity on its substrate, a recombinant synaptic protein synaptobrevin 2 derivative. By analyzing the modulation of the pore current caused by the specific BoNT-B-digested peptide as a marker, the presence of BoNT-B at a subnanomolar concentration was identified within minutes. The nanopore detector would fill the niche for a much needed rapid and highly sensitive detection of neurotoxins, and provide an excellent system to explore biophysical mechanisms for biopolymer transportation.
authors.affiliation: Department of Bioengineering and Dalton Cardiovascular Research Center, University of Missouri , Columbia, Missouri 65211, United States.
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.forename: Vedrana
authors.initials: V
authors.lastname: Montana
authors.forename: Vladimir
authors.initials: V
authors.lastname: Grubišić
authors.forename: Randy F
authors.initials: RF
authors.lastname: Stout
authors.forename: Vladimir
authors.initials: V
authors.lastname: Parpura
authors.forename: Li-Qun
authors.initials: LQ
authors.lastname: Gu
mesh_terms: D000818:Animals; D055162:Biocatalysis; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D019274:Botulinum Toxins, Type A / Q000737:chemistry*; D004063:Digestion; D007536:Isomerism; D058608:Nanopores; D010455:Peptides / Q000737:chemistry*; D051381:Rats; D050988:Vesicle-Associated Membrane Protein 2 / Q000737:chemistry*
chemical_list: D010455:Peptides; D050988:Vesicle-Associated Membrane Protein 2; C096323:rimabotulinumtoxinB; D019274:Botulinum Toxins, Type A
keywords: biosensor; botulinum; nanopore; neurotoxin; peptide; single molecules