Improved detection of botulinum type E by rational design of a new peptide substrate for endopeptidase-mass spectrometry assay.
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.
Department of Biotechnology, Israel Institute for Biological Research, Ness Ziona 7410001, Israel.
Osnat
O
Rosen
Department of Analytical Chemistry, Israel Institute for Biological Research, Ness Ziona 7410001, Israel.
Liron
L
Feldberg
Department of Analytical Chemistry, Israel Institute for Biological Research, Ness Ziona 7410001, Israel.
Sigalit
S
Gura
Department of Biotechnology, Israel Institute for Biological Research, Ness Ziona 7410001, Israel. Electronic address: ranz@iibr.gov.il.
Ran
R
Zichel
D000595:Amino Acid Sequence; D001905:Botulinum Toxins / Q000032:analysis* / Q000378:metabolism*; D015195:Drug Design; D007118:Immunoassay; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*
D010455:Peptides; D001905:Botulinum Toxins; C096326:botulinum toxin type E
Botulinum neurotoxin; Detection; Mass spectrometry; Peptide substrate; UPLC