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Title: α-Conotoxin Decontamination Protocol Evaluation: What Works and What Doesn’t

Abstract

Nine publically available biosafety protocols for safely handling conotoxin peptides were tested to evaluate their decontamination efficacy. Circular dichroism (CD) spectroscopy and mass spectrometry (MS) were used to assess the effect of each chemical treatment on the secondary and primary structure of α-CTx MII [L10V, E11A]. Of the nine decontamination methods tested, treatment with 1% (m/v) solution of the enzymatic detergent Contrex™ EZ resulted in a 76.8% decrease in α-helical content as assessed by the mean residue ellipticity at 222 nm, and partial peptide digestion was demonstrated using high performance liquid chromatography mass spectrometry (HPLC-MS). Additionally, treatment with 6% sodium hypochlorite (m/v) resulted in 80.5% decrease in α-helical content and complete digestion of the peptide. The Contrex™ EZ treatment was repeated with three additional α-conotoxins (α-CTxs), α-CTxs LvIA, ImI and PeIA, which verified the decontamination method was reasonably robust. These results support the use of either 1% Contrex™ EZ solution or 6% sodium hypochlorite in biosafety protocols for the decontamination of α- CTxs in research laboratories.

Authors:
 [1];  [2];  [3]
  1. Boise State Univ., ID (United States). Biomolecular Sciences Graduate Programs
  2. Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Biological Sciences Division
  3. Boise State Univ., ID (United States). Biomolecular Sciences Graduate Programs. Department of Chemistry and Biochemistry
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1406762
Report Number(s):
PNNL-SA-129342
Journal ID: ISSN 2072-6651; TOXIB7; 400904120
Grant/Contract Number:  
AC05-76RL01830
Resource Type:
Accepted Manuscript
Journal Name:
Toxins
Additional Journal Information:
Journal Volume: 9; Journal Issue: 9; Journal ID: ISSN 2072-6651
Publisher:
MDPI
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; conotoxin; select agent; circular dichrosim (CD) spectroscopy; mass spectrometry (MS); high performance liquid chromatography (HPLC); peptide decontamination; biosafety protocols

Citation Formats

Turner, Matthew, Cort, John, and McDougal, Owen. α-Conotoxin Decontamination Protocol Evaluation: What Works and What Doesn’t. United States: N. p., 2017. Web. doi:10.3390/toxins9090281.
Turner, Matthew, Cort, John, & McDougal, Owen. α-Conotoxin Decontamination Protocol Evaluation: What Works and What Doesn’t. United States. doi:10.3390/toxins9090281.
Turner, Matthew, Cort, John, and McDougal, Owen. Thu . "α-Conotoxin Decontamination Protocol Evaluation: What Works and What Doesn’t". United States. doi:10.3390/toxins9090281. https://www.osti.gov/servlets/purl/1406762.
@article{osti_1406762,
title = {α-Conotoxin Decontamination Protocol Evaluation: What Works and What Doesn’t},
author = {Turner, Matthew and Cort, John and McDougal, Owen},
abstractNote = {Nine publically available biosafety protocols for safely handling conotoxin peptides were tested to evaluate their decontamination efficacy. Circular dichroism (CD) spectroscopy and mass spectrometry (MS) were used to assess the effect of each chemical treatment on the secondary and primary structure of α-CTx MII [L10V, E11A]. Of the nine decontamination methods tested, treatment with 1% (m/v) solution of the enzymatic detergent Contrex™ EZ resulted in a 76.8% decrease in α-helical content as assessed by the mean residue ellipticity at 222 nm, and partial peptide digestion was demonstrated using high performance liquid chromatography mass spectrometry (HPLC-MS). Additionally, treatment with 6% sodium hypochlorite (m/v) resulted in 80.5% decrease in α-helical content and complete digestion of the peptide. The Contrex™ EZ treatment was repeated with three additional α-conotoxins (α-CTxs), α-CTxs LvIA, ImI and PeIA, which verified the decontamination method was reasonably robust. These results support the use of either 1% Contrex™ EZ solution or 6% sodium hypochlorite in biosafety protocols for the decontamination of α- CTxs in research laboratories.},
doi = {10.3390/toxins9090281},
journal = {Toxins},
number = 9,
volume = 9,
place = {United States},
year = {2017},
month = {9}
}

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