Details:

Alternative Title:
Anal Biochem
Publisher's site:
http://dx.doi.org/10.1016/j.ab.2012.09.021
Personal Author:
Wang, Dongxia ; Baudys, Jakub ; Ye, Yiming ; Rees, Jon C. ; Barr, John R. ; ... More +
Wang, Dongxia ; Baudys, Jakub ; Ye, Yiming ; Rees, Jon C. ; Barr, John R. ; Pirkle, James L. ; Kalb, Suzanne R. Less -
Description:
Botulinum neurotoxins (BoNTs) are a family of seven toxin serotypes that are the most toxic substances known to humans. Intoxication with BoNT causes flaccid paralysis and can lead to death if untreated with serotype-specific antibodies. Supportive care, including ventilation, may be necessary. Rapid and sensitive detection of BoNT is necessary for timely clinical confirmation of clinical botulism. Previously, our laboratory developed a fast and sensitive mass spectrometry (MS) method termed the Endopep-MS assay. The BoNT serotypes are rapidly detected and differentiated by extracting the toxin with serotype-specific antibodies and detecting the unique and serotype-specific cleavage products of peptide substrates that mimic the sequence of the BoNT native targets. To further improve the sensitivity of the Endopep-MS assay, we report here the optimization of the substrate peptide for the detection of BoNT/A. Modifications on the terminal groups of the original peptide substrate with acetylation and amidation significantly improved the detection of BoNT/A cleavage products. The replacement of some internal amino acid residues with single or multiple substitutions led to further improvement. An optimized peptide increased assay sensitivity 5-fold with toxin spiked into buffer solution or different biological matrices.
Subject:
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Acetylation
Amino Acid Sequence
Article
Botulinum Neurotoxin
Botulinum Toxins, Type A
Botulism
Detection
Endopeptidases
Immunoglobulin G
Mass Spectrometry
Molecular Sequence Data
Peptides
Peptide Substrate
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Substrate Specificity

Source:
Anal Biochem. 432(2):115-123.
Pubmed ID:
23017875
Pubmed Central ID:
PMC4618377
Document Type:
Journal Article
Funding:
CC999999/Intramural CDC HHS/United States
Collection(s):
CDC Public Access
Main Document Checksum:
[+]
urn:sha256:a2bc1346af54010464e2e2a485b2d0441bf5f2d6762b6b46d4c23ef7016a3550
File Type:

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