title: A new peptide substrate for enhanced botulinum neurotoxin type B detection by endopeptidase-liquid chromatography-tandem mass spectrometry/multiple reaction monitoring assay.
abstract: Botulinum neurotoxins (BoNTs) are the most toxic proteins in nature. Rapid and sensitive detection of BoNTs is achieved by the endopeptidase-mass spectrometry (Endopep-MS) assay. In this assay, BoNT cleaves a specific peptide substrate and the cleaved products are analyzed by MS. Here we describe the design of a new peptide substrate for improved detection of BoNT type B (BoNT/B) in the Endopep-MS assay. Our strategy was based on reported BoNT/B-substrate interactions integrated with analysis method efficiency considerations. Incorporation of the new peptide led to a 5-fold increased sensitivity of the assay both in buffer and in a clinically relevant human spiked serum.
authors.affiliation: Department of Biotechnology, Israel Institute for Biological Research, Ness Ziona 74100, Israel.
authors.forename: Osnat
authors.initials: O
authors.lastname: Rosen
authors.affiliation: Department of Analytical Chemistry, Israel Institute for Biological Research, Ness Ziona 74100, Israel.
authors.forename: Liron
authors.initials: L
authors.lastname: Feldberg
authors.affiliation: Department of Analytical Chemistry, Israel Institute for Biological Research, Ness Ziona 74100, Israel.
authors.forename: Sigalit
authors.initials: S
authors.lastname: Gura
authors.affiliation: Department of Biotechnology, Israel Institute for Biological Research, Ness Ziona 74100, Israel. Electronic address: ranz@iibr.gov.il.
authors.forename: Ran
authors.initials: R
authors.lastname: Zichel
mesh_terms: D000595:Amino Acid Sequence; D015374:Biosensing Techniques / Q000379:methods*; D019274:Botulinum Toxins, Type A / Q000097:blood / Q000378:metabolism*; D002853:Chromatography, Liquid; D010450:Endopeptidases / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D059748:Proteolysis; D053719:Tandem Mass Spectrometry
chemical_list: D010455:Peptides; C096323:rimabotulinumtoxinB; D010450:Endopeptidases; D019274:Botulinum Toxins, Type A
keywords: Botulinum neurotoxin; Detection; Mass spectrometry; Peptide substrate; UPLC