New generation protective peptide vaccine models: Development of bioconjugates of rabies viral peptides with various adjuvants

Researches which conducted in polyfunctional polymer systems provide new opportunities in the basic science and bioengineering fields. In this study, mechanism of interaction between synthetic polyelectrolytes and peptides, and physicochemical criterias for the occurence of functional biopolymer systems were investigated. To obtain disease germ and virus antigens (peptide, liopolysaccharide, etc.), produce them synthetically and thus develop vaccine lies on the basis of the new approaches and methods which are alternative for Pastor vaccine method. In this thesis, rabies virus protein's peptid epitope 31D (WAVYTRIMMNGGRLKRC) sequence which shows antigenic properties was synthesized with solid phase peptide synthesis methods using microwave energy in the peptide synthesizer (Liberty Microwave Assisted Solid Phase Peptide Synthesis Tool). Purified and characterizeid peptide's water-soluble polymer-peptide bioconjugates were synthesized in the presence of crosslinker '1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) with Polyacrilic acid (PAA) and Poly(N-vinyl-2-pyrrolidone-co-acrylic acid) [P(VP -co-AA)]. In this study, poly(N-vinyl-2-pyrrolidone-co-acrylic acid) copolymers which will be used as carrier molecule were synthesized and it's structure-function analysis were performed. Bioconjugates were synthesized as PAA polymer is constant, peptide has different rates (npep/nPAA), and peptide is constant, polymer has different rates (nPAA/npep), respectively. The characterization of prepared bioconjugates are made with HPLC, GPC (UV, refractive index, light scattering detectors), Zetasizer and FTIR. Rabies virus protein of 31D peptide epitope sequence's bioconjugates which P(VP-co-AA) is constant and peptide has different rates (npep / NPAA) were synthesized with two methods according to EDC activation method and its characterization were made with HPLC, GPC (UV, refractive index, light scattering detectors), Zetasizer and FTIR. Conjugates' cytotoxic effects are examined using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) method. Peptid loaded nanoparticles were formed with rabies virus of 31D sequence and Poly(lactic-co-glycolic acid) (PLGA). Zeta Sizer equepment was used for dimensional analysis and AFM and SEM equepments were used for morphological analysis. MTT cytotoxicity assay was performed for the examination bioavailability of peptide-loaded nanoparticles. FITC-loaded PLGA nanoparticles and both FITC and peptide loaded nanoparticles were synthesized to see the degredation of nanoparticles in florescence microscopy. Formation of new generation nanoparticles against rabies virus using nanotechnology which is one of the disciplines in the agenda of the last years holds promise for the development of vaccine prototypes in our future works. The results from this study are expected to be helped for formation of both straight chain and nano-structured peptide-polymer vaccine prototypes.