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Improved detection of botulinum type E by rational design of a new peptide substrate for endopeptidase–mass spectrometry assay
- Rosen, Osnat, Feldberg, Liron, Gura, Sigalit, Zichel, Ran
- Analytical biochemistry 2014 v.456 pp. 50-52
- botulinum toxin, cleavage (chemistry), in vitro studies, mass spectrometry, proteinases
- Botulinum neurotoxins (BoNTs) are the most toxic substances known to humans. Endopeptidase–mass spectrometry (Endopep–MS) is used as a specific and rapid in vitro assay to detect BoNTs. In this assay, immunocaptured toxin cleaves a serotype-specific peptide substrate, and the cleavage products are then detected by MS. To further improve the sensitivity of the assay, we report here the rational design of a new substrate peptide for the detection of botulinum neurotoxin type E (BoNT/E). Our strategy was based on previously reported structural interactions integrated with analysis method efficiency considerations. Integration of the newly designed substrate has led to a more than one order of magnitude increased sensitivity of the assay.