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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.
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Valérie Morineaux, Christelle Mazuet, Didier Hilaire, Julien Enche, Michel Popoff. Characterization of botulinum neurotoxin type A subtypes by immunocapture enrichment and liquid chromatography–tandem mass spectrometry. Analytical and Bioanalytical Chemistry, Springer Verlag, 2015, 407 (19), pp.5559 - 5570. ⟨10.1007/s00216-015-8707-1⟩. ⟨pasteur-01783532⟩

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