Identification of Snake Species by Toxin Mass Fingerprinting of Their Venoms

  • Reto Stöcklin
  • Dietrich Mebs
  • Jean-Claude Boulain
  • Pierre-Alain Panchaud
  • Henri Virelizier
  • Cécile Gillard-Factor
Part of the Methods in Molecular Biology™ book series (MIMB, volume 146)

Abstract

A new method is proposed to identify venomous snakes, which is based on electrospray ionization mass spectrometry (ESI-MS) detection and is demonstrated with Asiatic snake venoms from the controversial genus Naja (cobras). Appropriate combinations of chromatographic techniques and ESI-MS are used to analyze the crude venom of single specimens. Highly specific toxin mass maps, which can be used as a unique fingerprint for the systematic classification of the snake, are obtained; these results are compared with those obtained using standard samples and with the calculated molecular weights of characterized toxins. By off-line ESI-MS analysis of high-performance liquid chromatography (HPLC) fractions of two venom samples, one from Vietnam (undefined Naja sp.) and the other from Thailand (Naja kaouthia), it was found that both snakes belong to the same species, namely, Naja kaouthia. Using on-line liquid chromatography (LC)/ESI-MS, a direct analysis of crude venom from a single specimen of an unidentified white cobra from Thailand was performed. Two standard venom samples of Naja naja and Naja kaouthia were also analyzed using this improved strategy. By this approach, a peptide mass map of these three samples was obtained within a day and, in addition, an unambiguous systematic classification of the white cobra as Naja kaouthia was obtained. This method is able to identify clearly the origin and purity of crude or partially fractionated venom, which is an important advantage for medical use or in antivenom production.

Keywords

HPLC Acetonitrile Cysteine Electrophoresis Syringe 

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Copyright information

© Humana Press Inc. 2000

Authors and Affiliations

  • Reto Stöcklin
    • 1
  • Dietrich Mebs
    • 2
  • Jean-Claude Boulain
    • 3
  • Pierre-Alain Panchaud
    • 4
  • Henri Virelizier
    • 5
  • Cécile Gillard-Factor
    • 6
  1. 1.Atheris Laboratories,Research and DevelopmentGenevaSwitzerland
  2. 2.Zentrum der RechtsmedizinFrankfurtGermany
  3. 3.Département d’Ingénierie et d’Etude des ProtéinesCEA-SaclayFrance
  4. 4.Reptiles du Monde,GrandvauxSwitzerland
  5. 5.CEA SaclaySPEA-SAISCedexFrance
  6. 6.Département des Procédés d’Enrichissement,DCC/DPE/SPCP/LASOCEA-SaclayFrance

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