Preparation and Characterization of Agkistrodon Halys Venom Entrapped Chitosan Nanoparticles: Novel and Advanced Antigen Delivery and Adjuvant System

Background & Aims: In recent years, the feasibility of hydrophilic nanoparticles has been broadly investigated for use in drug delivery and therapeutic systems. Due to the problems of traditional adjuvants, in this study Agkistrodon halys (Ah) Snake venom was loaded in chitosan nanoparticles (CS NPs) in order to be used as an advanced adjuvant and antigen delivery system in antidote production industry. Methods: CS NPs containing Ah venom were prepared using ionic gelation and crosslinker method. In this study, polymer concentration, cross-linker concentration, particles size, particle size distribution, zeta potential, particles shape and surface characteristics, loading efficiency, loading capacity, and particles structure were optimized. Results: Optimum concentration achieved were chitosan 2mg/ml and cross-linker 1mg/ml and Agkistrodon Halys snake venum 500 mg/ml. Mean particle size, polydispersity index (PDI), and zeta potential of of venom-loaded CS NPs were respectively 129 nm, 0.418 and 41 mV. The loading capacity (LC) and loading efficiency (LE) were 82±1.9 and 88±2.1, respectively. Fourier transform infrared spectroscopy of nanoparticles showed between chitosan and crosslinker. An initial burst release and second slow release up to 96 hours was observed. Conclusion: The obtained results suggested that venom-loaded CS NPs could be a suitable alternative to conventional adjuvant for manufacturing antivenom.