This work used eleven Peruvian snake venoms (Bothrops andianus, Bothrops atrox, Bothrops barnetti, Bothrops castelnaudi, Bothriopsis chloromelas, Bothrocophias microphthalmus, Bothrops neuwiedi, Bothriopsis oligolepis, Bothriopsis peruviana, Bothrops pictus and Bothriopsis taeniata) to perform in vitro experimentation and determine its main characteristics. Hyaluronidase (HYAL), phospholipase A2 (PLA2), snake venom metalloproteinase (SVMP), snake venom serine protease (SVSP) and L-amino acid oxidase (LAAO) activities; toxicity by cell viability assays using MGSO3, VERO and HeLa cell lineages; and crossed immunoreactivity with Peruvian (PAV) and Brazilian (BAV) antibothropic polyvalent antivenoms, through ELISA and Western Blotting assays, were determined. Results show that the activities tested in this study were not similar amongst the venoms and each species present their own peculiarities, highlighting the diversity within Bothrops complex. All venoms were capable of reducing cell viability of all tested lineages. It was also demonstrated the crossed recognition of all tested venoms by both antivenoms.
Antivenins/pharmacology , Bothrops , Crotalid Venoms/toxicity , Animals , Blotting, Western , Brazil , Cell Line , Chlorocebus aethiops , Crotalid Venoms/enzymology , Crotalid Venoms/immunology , Enzyme-Linked Immunosorbent Assay , HeLa Cells , Humans , Hyaluronoglucosaminidase/metabolism , L-Amino Acid Oxidase/metabolism , Metalloproteases/metabolism , Peru , Phospholipases A2/metabolism , Serine Proteases/metabolism , Vero Cells
Mutalysin-II (mut-II) from Lachesis muta snake venom is an endopeptidase with hemorrhagic activity. A mAb against mutalysin-II that neutralized the hemorrhagic effect was produced previously. To identify the mAb epitopes, sets of 15-mer overlapping peptides covering the mut-II amino acid sequence were synthesized using the SPOT method and tested but failed to react with the mAb. Using a phage-display approach seventeen clones reactive with mAb were identified. Additional immunoassays with the peptides and mAb identified the QCTMDQGRLRCR, TCATDQGRLRCT, HCFHDQGRVRCA, HCTMDQGRLRCR and SCMLDQGRSRCR sequences as possible epitopes. Immunization of rabbits with these peptides induced antibodies that recognize mut-II and protected against the hemorrhagic effects of Lachesis venom.
Antibodies, Monoclonal/immunology , Crotalid Venoms/immunology , Hemorrhage/prevention & control , Metalloendopeptidases/immunology , Peptides/immunology , Amino Acid Sequence , Animals , Antibodies, Monoclonal/metabolism , Epitopes , Female , Hemorrhage/immunology , Metalloendopeptidases/metabolism , Molecular Sequence Data , Peptide Library , Peptides/metabolism , Rabbits , Vaccination
