Poster Presentation 11th Asia-Pacific Congress of the International Society on Toxinology 2021

Analysis by RP-HPLC of the protein profile of Bothrocophias myersi, Crotalus durissus and Bothrops asper from Colombian Andean Region (#101)

Stefano Scovino 1 , Karen Sarmiento 2 , Carlos Galvis 3 , Ana Lucía Castiblanco 1 , Fabio Aristizabal 4
  1. Laboratorio de Análisis Instrumental, Instituto de Biotecnología, Universidad Nacional de Colombia, Bogotá, Bogotá DC, Colombia
  2. Departamento de Ciencias Fisiológicas, Facultad de Medicina, Pontificia Universidad Javeriana, Bogotá, Bogotá DC, Colombia
  3. Colección de Fauna, Fundación Zoológica de Cali, Cali, Valle del Cauca, Colombia
  4. Departamento de Farmacia, Instituto de Biotecnología, Universidad Nacional de Colombia, Bogotá, Bogotá DC, Colombia

Snake venoms comprise a highly complex mixture of proteins, and there is also a high interspecific and intraspecific variability in their composition, even in the same region. Our aim was to compare the composition of the venoms of Bothrocophias myersi, Crotalus durissus, and Bothrops asper, snakes from the Colombian Andean region by Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC). The venoms were given to the research group under an agreement with Fundación Zoológica de Cali. The venoms pool was obtained by manual extraction, lyophilized and frozen. The venom protein was quantified by direct measurement with Nanodrop® 280 nm. The protein composition was established by RP-HPLC, using a Lichosper 100 RP, C18 column (250X4 mm) with a pore size of 5µm, as well as by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). The highest quantity of protein was found in the venom of B. myersi (108.6 mg/mL) followed by C. durissus (78.1 mg/mL) and B. asper (74.1 mg/mL). All venoms showed bands of 15 and 50 KDa by using SDS-PAGE. B. myersi venom chromatogram exhibited 16 peaks by RP-HPLC. We conclude that the composition of the three venoms is quite similar, being phospholipase A2 the common protein therein, and together with metalloproteinases they were the most abundant protein families in the venom of B. myersi. SDS-PAGE and RP-HPLC techniques allow a first approach to the profile of the venoms, which in turn could clarify the clinical syndrome produced

 

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