Development of a safe and effective vaccine is necessary to prevent RSV infection. Our project involved development of a multivalent vaccine for RSV by amplification of the antigenic regions of the RSV F, G and M2 genes which were subsequently cloned into a bacterial vector, pET-32. The recombinant clone was expressed from the transformed E. coli BL21 (DE3) cells purified and analyzed yielding a protein band of 38 kDa as predicted. The multivalent protein was encapsulated in chitosan to form protein encapsulated nanoparticle using the incorporation method. The encapsulated nanoparticles were analyzed for temperature stability, in vitro release of the protein, and also characterized using cytoviva bioimaging system as well as FTIR. The nanoencapsulated protein was stable up to a temperature of 1100C. The cytoviva bioimaging system revealed that the nanoparticles were irregular spherical shaped and the protein encapsulated nanoparticle appeared to be larger compared to the chitosan alone. The FTIR confirmed the chemical composition of the nanoparticle which consisted of polysaccharide, tripolyphosphate and protein. BALB/c female mice were immunized with the nanoencapsulated multivalent protein and the antibody response was compared to the mice vaccinated with live RSV or PBS The serum antibody response data revealed that mice immunized with the nanoencapsulated vaccine had the highest levels of antibody against RSV.
Keywords: RSV, Antibody, Vaccine, Recombinant DNA, Chitosan nanoparticle.
Sponsors: NSF-CREST, NIH-MBRS