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Toxins 2017, 9(9), 281; doi:10.3390/toxins9090281

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

1
Biomolecular Sciences Graduate Programs, Boise State University, Boise 83725, ID, USA
2
Department of Chemistry and Biochemistry, Boise State University, Boise 83725, ID, USA
3
Biological Sciences Division, Pacific Northwest National Laboratory, Richland 99354, WA, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Luis M. Botana
Received: 11 August 2017 / Revised: 4 September 2017 / Accepted: 9 September 2017 / Published: 14 September 2017
(This article belongs to the Collection Marine and Freshwater Toxins)
View Full-Text   |   Download PDF [1188 KB, uploaded 14 September 2017]   |  

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. View Full-Text
Keywords: conotoxin; select agent; circular dichrosim (CD) spectroscopy; mass spectrometry (MS); high performance liquid chromatography (HPLC); peptide decontamination; biosafety protocols conotoxin; select agent; circular dichrosim (CD) spectroscopy; mass spectrometry (MS); high performance liquid chromatography (HPLC); peptide decontamination; biosafety protocols
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Turner, M.W.; Cort, J.R.; McDougal, O.M. α-Conotoxin Decontamination Protocol Evaluation: What Works and What Doesn’t. Toxins 2017, 9, 281.

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