title: Characterization of botulinum neurotoxin type A subtypes by immunocapture enrichment and liquid chromatography-tandem mass spectrometry.
abstract: Botulinum neurotoxins (BoNT) are divided into seven toxinotypes based on their immunological properties and each toxinotype contains several subtypes according to their amino acid sequences. Here, we designed a mass spectrometry method able to identify BoNT/A subtypes in complex matrices including crude culture supernatants, food, and environmental samples. Peptides from BoNT light chain (L) specific to the subtypes BoNT/A1 to A3 and BoNT/A5 to A8 were identified. The method consists of an immunocapture step with antibodies specific to BoNT/A L chains followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) on a triple quadrupole mass spectrometer (QqQ) in multiple reaction monitoring (MRM) mode. BoNT/A subtypes were correctly identified in culture supernatants and in tap water or orange juice samples with a limit of detection of 20 to 150 mouse lethal doses (MLD) and with a lower sensitivity in serum samples.
authors.affiliation: Direction Générale de l'Armement, DGA Maîtrise NRBC, 91710, Vert-le-Petit, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Morineaux
authors.forename: Christelle
authors.initials: C
authors.lastname: Mazuet
authors.forename: Didier
authors.initials: D
authors.lastname: Hilaire
authors.forename: Julien
authors.initials: J
authors.lastname: Enche
authors.forename: Michel R
authors.initials: MR
authors.lastname: Popoff
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019274:Botulinum Toxins, Type A / Q000737:chemistry*; D002853:Chromatography, Liquid / Q000379:methods*; D005260:Female; D005506:Food Contamination; D057230:Limit of Detection; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D019274:Botulinum Toxins, Type A