PHYSICO-CHEMICAL STUDIES OF CROTOXIN ISOLATED FROM RATTLESNAKE VENOM ADSORBED TO SILICA NANOPARTICLES FOR POTENTIAL USE IN THE PRODUCTION OF NEW ANTIVENOMS

CONTI, FLORENCIA; GIORGI, EXEQUIEL; DIAZ, MARIA EUGENIA; DE MARZI, MAURICIO; RODRIGUEZ, JUAN PABLO; BAUDOU, FEDERICO

Mar del Plata

Congreso; Reunión Anual de Sociedades de Biociencias; 2022

The accident produced by venomous snakes, ophidism, is a globallyneglected health problem. In Argentina, Crotalus durissus terrificus(southern rattlesnake) is one of the species that generates the mostaccidents. Its venom (V) contains mainly crotoxin (CTX), responsible for the high lethality. On the other hand, the use of nanoparticlesopens a range of possibilities within the medical field given its potential therapeutic and immunomodulatory use. Therefore, the interestin transporting V toxins in the production of new antivenoms (AV).In this work, we isolate CTX by fast protein liquid chromatography(FPLC) and generate nanovenoms (NVs), complex formed by CTXadsorbed to silica nanoparticles (SiNPs) of 150 nm in diameter ofboth charges. For this, 1 ml of CTX was mixed under stirring with 10mg of SiNPs (+/-). The charge and size were corroborated by potential Z and DLS (dynamic light scattering) respectively. NVs were alsomicrophotographed by Transmission Electron Microscope (TEM)and analyzed by Fourier Transform Infrared (FT-IR). Additionally, westudied the enzymatic activity exerted by CTX present in NVs by hemolysis radial test. The results obtained by FPLC allowed to isolateCTX, the maximum peak (11th fraction), then purified and concentrated by dialysis and lyophilized (2 mg/ml PBS 10 mM). ThroughTEM we can observe on NVs the protein halo corresponding to thepresence of CTX, and the FT-IR spectra obtained show the peak(abs 510 cm-1) corresponding to H-S-H bounds characteristic ofCTX. Finally, it was found that NVs retain their enzymatic activitythanks to the halo produced in the hemolysis radial test. The use ofthese complexes studied here at the physical-chemical level, wherethe activity of CTX adsorbed to NVs is also preserved, it allows us tocontinue perfecting their possible use as adjuvants in the productionof new generation AV.