title: Vintage venoms: proteomic and pharmacological stability of snake venoms stored for up to eight decades.
abstract: For over a century, venom samples from wild snakes have been collected and stored around the world. However, the quality of storage conditions for "vintage" venoms has rarely been assessed. The goal of this study was to determine whether such historical venom samples are still biochemically and pharmacologically viable for research purposes, or if new sample efforts are needed. In total, 52 samples spanning 5 genera and 13 species with regional variants of some species (e.g., 14 different populations of Notechis scutatus) were analysed by a combined proteomic and pharmacological approach to determine protein structural stability and bioactivity. When venoms were not exposed to air during storage, the proteomic results were virtually indistinguishable from that of fresh venom and bioactivity was equivalent or only slightly reduced. By contrast, a sample of Acanthophis antarcticus venom that was exposed to air (due to a loss of integrity of the rubber stopper) suffered significant degradation as evidenced by the proteomics profile. Interestingly, the neurotoxicity of this sample was nearly the same as fresh venom, indicating that degradation may have occurred in the free N- or C-terminus chains of the proteins, rather than at the tips of loops where the functional residues are located. These results suggest that these and other vintage venom collections may be of continuing value in toxin research. This is particularly important as many snake species worldwide are declining due to habitat destruction or modification. For some venoms (such as N. scutatus from Babel Island, Flinders Island, King Island and St. Francis Island) these were the first analyses ever conducted and these vintage samples may represent the only venom ever collected from these unique island forms of tiger snakes. Such vintage venoms may therefore represent the last remaining stocks of some local populations and thus are precious resources. These venoms also have significant historical value as the Oxyuranus venoms analysed include samples from the first coastal taipan (Oxyuranus scutellatus) collected for antivenom production (the snake that killed the collector Kevin Budden), as well as samples from the first Oxyuranus microlepidotus specimen collected after the species' rediscovery in 1976. These results demonstrate that with proper storage techniques, venom samples can retain structural and pharmacological stability. This article is part of a Special Issue entitled: Proteomics of non-model organisms.
  
  
  CONCLUSIONS
authors.affiliation: Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Clémence
authors.initials: C
authors.lastname: Jesupret
authors.affiliation: Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Kate
authors.initials: K
authors.lastname: Baumann
authors.affiliation: Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4072, Australia; Institute for Molecular Bioscience, University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Timothy N W
authors.initials: TN
authors.lastname: Jackson
authors.affiliation: Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4072, Australia; HEJ Research Institute of Chemistry, International Centre for Chemical and Biological Sciences (ICCBS), University of Karachi, Karachi 75270, Pakistan.
authors.forename: Syed Abid
authors.initials: SA
authors.lastname: Ali
authors.affiliation: Monash Venom Group, Department of Pharmacology, Monash University, Clayton, Victoria 3800, Australia.
authors.forename: Daryl C
authors.initials: DC
authors.lastname: Yang
authors.affiliation: Australian Venom Research Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Victoria 3010, Australia; Monash Venom Group, Department of Pharmacology, Monash University, Clayton, Victoria 3800, Australia.
authors.forename: Laura
authors.initials: L
authors.lastname: Greisman
authors.affiliation: Australian Venom Research Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Victoria 3010, Australia.
authors.forename: Larissa
authors.initials: L
authors.lastname: Kern
authors.affiliation: Australian Venom Research Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Victoria 3010, Australia.
authors.forename: Jessica
authors.initials: J
authors.lastname: Steuten
authors.affiliation: Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Mahdokht
authors.initials: M
authors.lastname: Jouiaei
authors.affiliation: Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4072, Australia; Alistair Reid Venom Research Unit, Liverpool School of Tropical Medicine, Liverpool L3 5QA, UK; Molecular Ecology and Evolution Group, School of Biological Sciences, Bangor University, Bangor LL57 2UW, UK.
authors.forename: Nicholas R
authors.initials: NR
authors.lastname: Casewell
authors.affiliation: Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4072, Australia; Institute for Molecular Bioscience, University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Eivind A B
authors.initials: EA
authors.lastname: Undheim
authors.affiliation: Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Ivan
authors.initials: I
authors.lastname: Koludarov
authors.affiliation: Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Jordan
authors.initials: J
authors.lastname: Debono
authors.affiliation: Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Dolyce H W
authors.initials: DH
authors.lastname: Low
authors.affiliation: Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Sarah
authors.initials: S
authors.lastname: Rossi
authors.affiliation: Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Nadya
authors.initials: N
authors.lastname: Panagides
authors.affiliation: Monash Venom Group, Department of Pharmacology, Monash University, Clayton, Victoria 3800, Australia.
authors.forename: Kelly
authors.initials: K
authors.lastname: Winter
authors.affiliation: Department of Paediatrics, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, Victoria 3000, Australia; Murdoch Childrens Research Institute, Parkville, Victoria 3052, Australia.
authors.forename: Vera
authors.initials: V
authors.lastname: Ignjatovic
authors.affiliation: Department of Paediatrics, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, Victoria 3000, Australia.
authors.forename: Robyn
authors.initials: R
authors.lastname: Summerhayes
authors.affiliation: Institute for Molecular Bioscience, University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Alun
authors.initials: A
authors.lastname: Jones
authors.affiliation: School of Chemistry and Molecular Biosciences, University of Queensland, St. Lucia, Qld 4072, Australia.
authors.forename: Amanda
authors.initials: A
authors.lastname: Nouwens
authors.affiliation: Venom Supplies, Tanunda, SA, Australia.
authors.forename: Nathan
authors.initials: N
authors.lastname: Dunstan
authors.affiliation: Monash Venom Group, Department of Pharmacology, Monash University, Clayton, Victoria 3800, Australia.
authors.forename: Wayne C
authors.initials: WC
authors.lastname: Hodgson
authors.affiliation: Australian Venom Research Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Victoria 3010, Australia.
authors.forename: Kenneth D
authors.initials: KD
authors.lastname: Winkel
authors.affiliation: Department of Paediatrics, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, Victoria 3000, Australia.
authors.forename: Paul
authors.initials: P
authors.lastname: Monagle
authors.affiliation: Venom Evolution Lab, School of Biological Sciences, University of Queensland, St. Lucia, Queensland 4072, Australia; Institute for Molecular Bioscience, University of Queensland, St. Lucia, Queensland 4072, Australia. Electronic address: bgfry@uq.edu.au.
authors.forename: Bryan Grieg
authors.initials: BG
authors.lastname: Fry
mesh_terms: D004546:Elapid Venoms / Q000737:chemistry*; D011309:Preservation, Biological; D055550:Protein Stability; D040901:Proteomics / Q000379:methods*; D013997:Time Factors
chemical_list: D004546:Elapid Venoms; C072139:Notechis 11'2
keywords: 1D gel; Coastal taipan; Death adder; Mass spectrometry; Protein; Snake venom detection; Stability studies; Toxinology; Venom