A new peptide substrate for enhanced botulinum neurotoxin type B detection by endopeptidase-liquid chromatography-tandem mass spectrometry/multiple reaction monitoring assay

Anal Biochem. 2015 Mar 15:473:7-10. doi: 10.1016/j.ab.2014.09.016. Epub 2014 Sep 30.

Abstract

Botulinum neurotoxins (BoNTs) are the most toxic proteins in nature. Rapid and sensitive detection of BoNTs is achieved by the endopeptidase-mass spectrometry (Endopep-MS) assay. In this assay, BoNT cleaves a specific peptide substrate and the cleaved products are analyzed by MS. Here we describe the design of a new peptide substrate for improved detection of BoNT type B (BoNT/B) in the Endopep-MS assay. Our strategy was based on reported BoNT/B-substrate interactions integrated with analysis method efficiency considerations. Incorporation of the new peptide led to a 5-fold increased sensitivity of the assay both in buffer and in a clinically relevant human spiked serum.

Keywords: Botulinum neurotoxin; Detection; Mass spectrometry; Peptide substrate; UPLC.

MeSH terms

  • Amino Acid Sequence
  • Biosensing Techniques / methods*
  • Botulinum Toxins, Type A / blood
  • Botulinum Toxins, Type A / metabolism*
  • Chromatography, Liquid
  • Endopeptidases / metabolism*
  • Humans
  • Molecular Sequence Data
  • Peptides / chemistry
  • Peptides / metabolism*
  • Proteolysis*
  • Tandem Mass Spectrometry*

Substances

  • Peptides
  • rimabotulinumtoxinB
  • Endopeptidases
  • Botulinum Toxins, Type A