Vaccine Research (May 2017)
Preparation and characterization of PLGA Nanoparticles containing recombinant core-NS3 Fusion protein of hepatitis C virus as a nano-vaccine candidate
Abstract
Introduction: Hepatitis C virus (HCV) is one of the most serious causes of cirrhosis, liver cancer and ultimately death, worldwide. The new direct-acting drugs are not accessible for many patients around the world and progress toward new therapeutic and anaphylactic vaccines design has not been fast enough. This study was aimed to prepare and assess a recombinant fusion protein core-NS3 (rC-N) of HCV with the accompaniment of d,l-polylactide-co-glycolide nanoparticles (PLGA NPs) as a nano-conjugated vaccine candidate. Methods: The rC-N protein containing the first domain of core and middle region of NS3 (residues 1095-1384) was loaded into PLGA NPs (rC-N/PLGA NPs) by NHS and DDC (equimolar: 0.5 mM) as conjugating agents. The morphology and average of surface roughness (Ra) of PLGA NPs and rC-N/PLGA NPs were demonstrated by atomic force microscope (AFM). The particle sizes, polydispersity index (PDI) and zeta potential were measured by Zetasizer. Results: The morphology of the nanoparticles was spherical and their surface Ra was measured to be 7.630 nm for PLGA NPs and 15.72 nm for rC-N/PLGA NPs. The average size (160.4 nm), zeta potential (-37.6 mV) and PDI (0.227) were also obtained for rC-N/ PLGA NPs. Conclusion: The surface Ra value of rC-N/PLGA NPs (15.72 nm) which was twice more than PLGA NPs (7.630 nm) confirmed a successful conjugation. The stability of nanoparticles behavior in the colloid was confirmed by the absolute value of zeta potential (|-37.6|= 37.6 mV) of rC-N/PLGA NPs. The spherical morphology, average size 30 mV, PDI < 0.5 were confirmatory indications that rC-N/PLGA NPs could be considered as vaccine candidate. The rC-N/PLGA NPs construct should be further evaluated via in-vivo challenges and demonstration of targeted delivery to the antigen presenting cells.
