2005
DOI: 10.3201/eid1110.041279
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Botulinum Neurotoxin Detection and Differentiation by Mass Spectrometry

Abstract: A new rapid, mass spectrometry-based method to detect and differentiate botulinal neurotoxins is described.

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Cited by 161 publications
(153 citation statements)
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“…The sample was previously determined to contain BoNT A by the Endopep-MS assay 22 and the mouse assay and was subjected to MESID to obtain further information on the toxin subtype and variant on protein level. The toxin was extracted with antibody-coated beads and three identical samples were loaded into a 1D SDS gel ( Figure S3).…”
Section: Subtyping Of a Bont-contaminated Carrot Juice Samplementioning
confidence: 99%
“…The sample was previously determined to contain BoNT A by the Endopep-MS assay 22 and the mouse assay and was subjected to MESID to obtain further information on the toxin subtype and variant on protein level. The toxin was extracted with antibody-coated beads and three identical samples were loaded into a 1D SDS gel ( Figure S3).…”
Section: Subtyping Of a Bont-contaminated Carrot Juice Samplementioning
confidence: 99%
“…A floating consensus was derived as shown and was used to design two additional longer substrates (Table 1). LF-3 consists of the 11-amino acid core of LF-1 with an additional 3 amino (N)-terminal and 7 C-terminal amino acids that match the consensus, except for arginine (R 19 ), which was included for MS detection. LF-4 also includes the core, but extended sequences do not directly match the floating consensus.…”
Section: Resultsmentioning
confidence: 99%
“…Several internal residues and the terminal structure of this peptide were modified from the corresponding sequence of the SNAP-25 185-260 based on previous reported study (Schmidt and Bostian 1997) and experience gained during the initial development of the assay (Barr, Moura et al 2005;Boyer, Moura et al 2005). To evaluate whether higher sensitivities of the assay could be accomplished by further optimization of this substrate, we re-examined its function and primary structure.…”
Section: Terminal Modificationsmentioning
confidence: 99%
“…To evaluate the performance of the new substrate in the assay for detecting BoNT/A present in complex samples, we evaluated the specific cleavage of Pep-21 using BoNT/A toxin spiked into four common sample matrices: serum, stool extract, cell culture supernatant, and milk. The assays were conducted following the general procedure of the Endopep-MS method for clinical samples (Barr, Moura et al 2005;Kalb, Moura et al 2006;Wang, Baudys et al 2011). The toxin was first enriched from a spiked matrix by BoNT/A-specific antibody immobilized on magnetic beads followed by the cleavage reaction in a solution containing toxin bound to beads, Pep-21 and other reaction components.…”
Section: Complex Biological Matrixmentioning
confidence: 99%
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