Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 407, Issue 19, Pages 5559-5570Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-015-8707-1
Keywords
Botulinum neurotoxin; Clostridium botulinum; Subtyping; Mass spectrometry
Funding
- Direction Generale de l'Armement
- Institut Pasteur
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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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