Liquid chromatography - mass spectrometric analysis of botulinum neurotoxin serotype A light chain assay mixtures


  1. Chan, N.W.C.
  2. Tang, T.
  3. Lee, W.E.
  4. Gebremedhin, M.G.
  5. Mah, D.C.W.
Corporate Authors
Defence R&D Canada - Suffield, Ralston ALTA (CAN)
Botulinum neurotoxin is the most toxic natural, known substance that can cause poisoning with no effective cure. Botulinum neurotoxin serotype A (BoNT/A) is the most common and deadly among all seven serotypes (A through G). There is a need to access effective countermeasures against BoNT. One approach is to develop small molecule inhibitors against the toxin’s enzymatic active site, i.e the light chain (LC) domain. A high performance liquid chromatography coupled with mass spectrometry (HPLC-MS) assay was developed to separate the substrate and product peptides of the BoNT/A LC enzymatic assay. The HPLC-MS assay allows us to measure the amount of substrate and product peptides in order to determine the inhibitory effects of specific small molecule compounds on BoNT/A LC. The results were complementary to those obtained by fluorescence resonance energy transfer, the high throughput assay developed for BoNT/A LC previously. Potential inhibitors of BoNT/A LC were compiled for further comprehensive studies in computational molecular modeling, dose response and efficacy studies in cell and tissue assays.

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Botulinum neurotoxin;off-patent;marketed drugs;medical countermeasures
Report Number
DRDC-SUFFIELD-TM-2010-219 — Technical Memorandum
Date of publication
01 Dec 2010
Number of Pages
Electronic Document(PDF)

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