"Physico-chemical studies of crotoxin isolated from rattlesnake venom adsorbed to silica nanoparticles for potential use in the production of new antivenoms"

MARIA EUGENIA DIAZ; FEDERICO G. BAUDOU; FLORENCIA S. CONTI; EXEQUIEL GIORGI; MAURICIO DE MARZI; JUAN PABLO RODRÍGUEZ

Mar del Plata

Congreso; Reunión Anual de Sociedades de Biociencia 2022; 2022

Reunion conjunta: SAIC, SAI, SAFIS

The accident produced by venomous snakes, ophidism, is a globally neglected health problem. In Argentina, Crotalus durissus terrificus (southern rattlesnake) is one of the species that generates the most accidents. Its venom (V) contains mainly crotoxin (CTX), responsible for the high lethality. On the other hand, the use of nanoparticles opens a range of possibilities within the medical field given its potential therapeutic and immunomodulatory use. Therefore, the interest in 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 CTX adsorbed to silica nanoparticles (SiNPs) of 150 nm in diameter of both charges. For this, 1 ml of CTX was mixed under stirring with 10 mg of SiNPs (+/-). The charge and size were corroborated by potential Z and DLS (dynamic light scattering) respectively. NVs were also microphotographed by Transmission Electron Microscope (TEM) and analyzed by Fourier Transform Infrared (FT-IR). Additionally, we studied the enzymatic activity exerted by CTX present in NVs by hemolysis radial test. The results obtained by FPLC allowed to isolate CTX, the maximum peak (11th fraction), then purified and concentrated by dialysis and lyophilized (2 mg/ml PBS 10 mM). Through TEM we can observe on NVs the protein halo corresponding to the presence of CTX, and the FT-IR spectra obtained show the peak (abs 510 cm-1) corresponding to H-S-H bounds characteristic of CTX. Finally, it was found that NVs retain their enzymatic activity thanks to the halo produced in the hemolysis radial test. The use of these complexes studied here at the physical-chemical level, where the activity of CTX adsorbed to NVs is also preserved, it allows us to continue perfecting their possible use as adjuvants in the production of new generation AV.