α-Conotoxin Decontamination Protocol Evaluation: What Works and What Doesn’t
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 structur...
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doaj-3f86750f956a456a937db7627dc046542020-11-24T23:06:01ZengMDPI AGToxins2072-66512017-09-019928110.3390/toxins9090281toxins9090281α-Conotoxin Decontamination Protocol Evaluation: What Works and What Doesn’tMatthew W. Turner0John R. Cort1Owen M. McDougal2Biomolecular Sciences Graduate Programs, Boise State University, Boise 83725, ID, USABiological Sciences Division, Pacific Northwest National Laboratory, Richland 99354, WA, USADepartment of Chemistry and Biochemistry, Boise State University, Boise 83725, ID, USANine 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.https://www.mdpi.com/2072-6651/9/9/281conotoxinselect agentcircular dichrosim (CD) spectroscopymass spectrometry (MS)high performance liquid chromatography (HPLC)peptide decontaminationbiosafety protocols |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Matthew W. Turner John R. Cort Owen M. McDougal |
spellingShingle |
Matthew W. Turner John R. Cort Owen M. McDougal α-Conotoxin Decontamination Protocol Evaluation: What Works and What Doesn’t Toxins conotoxin select agent circular dichrosim (CD) spectroscopy mass spectrometry (MS) high performance liquid chromatography (HPLC) peptide decontamination biosafety protocols |
author_facet |
Matthew W. Turner John R. Cort Owen M. McDougal |
author_sort |
Matthew W. Turner |
title |
α-Conotoxin Decontamination Protocol Evaluation: What Works and What Doesn’t |
title_short |
α-Conotoxin Decontamination Protocol Evaluation: What Works and What Doesn’t |
title_full |
α-Conotoxin Decontamination Protocol Evaluation: What Works and What Doesn’t |
title_fullStr |
α-Conotoxin Decontamination Protocol Evaluation: What Works and What Doesn’t |
title_full_unstemmed |
α-Conotoxin Decontamination Protocol Evaluation: What Works and What Doesn’t |
title_sort |
α-conotoxin decontamination protocol evaluation: what works and what doesn’t |
publisher |
MDPI AG |
series |
Toxins |
issn |
2072-6651 |
publishDate |
2017-09-01 |
description |
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. |
topic |
conotoxin select agent circular dichrosim (CD) spectroscopy mass spectrometry (MS) high performance liquid chromatography (HPLC) peptide decontamination biosafety protocols |
url |
https://www.mdpi.com/2072-6651/9/9/281 |
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