title: Botulinum neurotoxin detection and differentiation by mass spectrometry.
abstract: Botulinum neurotoxins (BoNTs) are proteases that cleave specific cellular proteins essential for neurotransmitter release. Seven BoNT serotypes (A-G) exist; 4 usually cause human botulism (A, B, E, and F). We developed a rapid, mass spectrometry-based method (Endopep-MS) to detect and differentiate active BoNTs A, B, E, and F. This method uses the highly specific protease activity of the toxins with target peptides specific for each toxin serotype. The product peptides derived from the endopeptidase activities of BoNTs are detected by matrix-assisted laser-desorption ionization time-of-flight mass spectrometry. In buffer, this method can detect toxin equivalents of as little as 0.01 mouse lethal dose (MLD)50 and concentrations as low as 0.62 MLD50/mL. A high-performance liquid chromatography-tandem mass spectrometry method for quantifying active toxin, where the amount of toxin can be correlated to the amount of product peptides, is also described.
authors.affiliation: Centers for Disease Control and Prevention, Atlanta, Georgia 30341, USA. jbarr@cdc.gov
authors.forename: John R
authors.initials: JR
authors.lastname: Barr
authors.forename: Hercules
authors.initials: H
authors.lastname: Moura
authors.forename: Anne E
authors.initials: AE
authors.lastname: Boyer
authors.forename: Adrian R
authors.initials: AR
authors.lastname: Woolfitt
authors.forename: Suzanne R
authors.initials: SR
authors.lastname: Kalb
authors.forename: Antonis
authors.initials: A
authors.lastname: Pavlopoulos
authors.forename: Lisa G
authors.initials: LG
authors.lastname: McWilliams
authors.forename: Jurgen G
authors.initials: JG
authors.lastname: Schmidt
authors.forename: Rodolfo A
authors.initials: RA
authors.lastname: Martinez
authors.forename: David L
authors.initials: DL
authors.lastname: Ashley
mesh_terms: D000595:Amino Acid Sequence; D001905:Botulinum Toxins / Q000737:chemistry / Q000145:classification* / Q000378:metabolism*; D019274:Botulinum Toxins, Type A / Q000737:chemistry / Q000378:metabolism; D003014:Clostridium botulinum / Q000145:classification / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000737:chemistry / Q000145:classification* / Q000378:metabolism*; D012703:Serotyping; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D009498:Neurotoxins; C096323:rimabotulinumtoxinB; D010450:Endopeptidases; D001905:Botulinum Toxins; D019274:Botulinum Toxins, Type A; C096326:botulinum toxin type E; C096321:botulinum toxin type F