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Intraspecies variability in Vipera ammodytes ammodytes venom related to its toxicity and immunogenic potential
Authors:Halassy Beata  Brgles Marija  Habjanec Lidija  Balija Maja Lang  Kurtović Tihana  Marchetti-Deschmann Martina  Križaj Igor  Allmaier Günter
Affiliation:Research and Development Department, Institute of Immunology, Inc., Rockefellerova 10, HR-10 000 Zagreb, Croatia. bhalassy@imz.hr
Abstract:Vipera ammodytes is the most venomous European snake, whose venom has been used as antigen for immunization of antivenom-producing animals. Same as venom of any other snake, it is a complex mixture of proteins, peptides and other compounds which biochemical and pharmacological variability has been demonstrated at interspecies and intraspecies level. In this work we demonstrated intraspecific variability between 8 venom production batches using both the conventional and the new methodology. Moreover, in contrast to the literature on different venoms' variability, for the first time we were able to select those biochemical differences that are related to and give information on the venom's toxicity and immunogenicity. We have shown that methods quantifying ammodytoxin (the most toxic compound identified so far in the Vipera ammodytes ammodytes venom) content of the venom clearly distinguish between high and low immunogenic venoms.
Keywords:Atx, ammodytoxin   BSA, bovine serum albumin   CGE, capillary gel electrophoresis   CIM, convective interaction media   CM, carboxymethyl   DAD, photodiode array detection   EF, elution fraction   ELISA, enzyme linked immunosorbent assay   HPLC, high performance liquid chromatography   HRP, horseradish peroxidase   KR, Krapina (Croatian region)   LC-MS, liquid chromatography mass spectrometry   LD50, median lethal dose   MHD, minimum hemorrhagic dose   MND, minimum necrotic dose   OPD, o-phenylenediamine dihydrochloride   PLA, phospholipase activity   SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis   SE, standard error   SL, Slunj (Croatian region)   Vaa, Vipera ammodytes ammodytes   WFI, water for injection
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