Synthesis of antigenic synthetic peptide of Canine parvovirus and development of bioconjugates

The development of new and effective polymeric systems and the preparation-application of specific controlled delivery formulations, long-action peptide carrier extensively studied subject in biotechnolgy with the aim of further applications of drug and vaccine. Interaction of polymers with peptide, protein ect. biomolecules is great importance for understanding the action mechanism of biopolymers in organism and development of bioconjugation technology. Coupling of peptides to carrier molecules is required to antibody production against weak immunogenic peptides. Activation of carboxyl groups of water soluble biopolymers and covalent binding of peptides amino groups is basis of bioconjugation reaction of antigenic synthetic peptide of Canine Parvovirus which subject of this thesis.In the first part of this PhD. thesis W-7L20 synthetic peptide of Canine Parvovirus is synthezied by Solid Phase Peptide Synthesis Method and bioconjugated to PAA polymer. Furthermore commercially purchased W-1L19 synthetic peptide-Polymer bioconjugates with polyacrylic acid, poly(N-vinyl-2-pyrrolidone-co-acrylic acid), carboxymethylcelulase synthesized in the presence of ?1-ethyl-3-3-dimethyl aminopropyl karbodiimid? cross-linker. Bioconjugates were synthesized different ratio of peptides for one polymer molecule (npep/npol) and different ratio of polymer for one peptide molecule (npol/npep). After the syntesis step, peptide-polymer bioconjugates comperatively analyzed with peptide and polymers, for investigation of formation mechanism, electrical charge and three dimensional structure by using HPLC, GPC, Zetasizer and Fluorescence Spectrophotometer.In the second part of this PhD. thesis only Canine parvovirus synthetic peptide and peptide+fluorescein isothiocyanate (FITC) loaded Poly(lactide-co-glicolide)acid (PL-GA) nanoparticles produced. After the analyses of produced nanoparticle?s size and zeta potential analyses by Zetasizer, the chromatograms receipt by GPC comperatively by PL-GA polymer. After performed FT-IR analyses of nanoparticles, AFM and SEM analysed were done. Peptide loading capacity and release properties of nanoparticles analyzed by AFM and UV Spectrophotometer. Cytotoxic properties of produced nanoparticles was evaluated against mouse fibroblast cells (L-929) in comparison with peptide.Information to be obtained within the scope of this thesis is to shed light on the direction of, to obtain specific antibodies against bloody diarrhea which caused by Canine parvovirus, generation of diagnostic kits and biosensors and development of synthetic vaccines will be a pre-study.In addition, nanoparticles produced in this thesis are promising carriers for controlled release of antigenic peptides and for the development of biotechnological vaccines based on this.