Purification Of DHV-Ⅰ VP1Monoclonal Antibody And Identification Of Its Antigen Epitope

Duck viral hepatitis (DVH) is an acute, highly contagious, viral disease of ducklings. The maincharacterization of DVH is hepatitis. The agent of DVH is duck hepatitis virus (DHV). Three distinctserotypes have been detected, termed as DHV-I, DHV-II and DHV-III. No cross-reactive are detectedamong the three serotypes. DHV-I which divided into A, B and C three genotypes is the most widelydistributed of the three serotypes. Mainly in the following3-weeks cheeper duck spread rapidly andhigh mortality. In our country the outbreak of the duck hepatitis is mainly caused by the DHV-I. Onceinfected, the mortality of cheeper duck was almost100%. DHV-1is the most important and severe one.It is of economic importance in all duck-raising areas of the world.This study recovers2strains of cryopreserved positive hybridoma2D9,2D10respectively, andexpanding of the trainings are carried out for preparation of McAbs. Prepared2D9and2D10are used tobiological elutriation as target molecules by the use of random phage display12peptide library. Afterthree rounds of elutriation and affinity screening, and the corresponding get8positive phage cloningsrespectively, the sequencing and analysis, finally respectively get peptides Cpep1:-LPAPTS-and Cpep2:-RSND-. The synthetic peptides, through the competitive inhibition test, affinity test and ELISAdetection, proof of synthetic peptide can restrain the combination of the VP1protein and McAb,confirm Cpep1:-LPAPTS-and Cpep2:-RSND-are two antigen epitope for DHV-I VP1protein.This study succeeded in purification of monoclonal antibody two strains of high purity2D9and2D10, using phage display technology and the success of the identification of two strains of monoclonalantibody2D9,2D10antigen epitope, Recognizing its antigen structure, to study table a biologicalfunction, the protein antigen structure foundation. Due to different basic no cross between serum typeduck hepatitis virus protection, so the appraisal of the antigen epitope can also be designed for scientificand effective genetic engineering vaccine, especially epitope-based vaccines, the development of thecorresponding diagnostic reagents, scout ahead of time. For provide scientific basis for subsequentresearch and biological materials. These for disease prevention and control of the duck industry in ourcountry and related industry development plays an important practical significance.This study has mastered the purified antigen identification technology, phage display technology,in order to further explore CsdA protein structure and function in the mycobacterium tuberculosisprovides a convenient means of research, to seek treatment for drug targets for type I duck thereresearch provides an important reference material, and on the prevention and control of type I duck there has important theoretical significance.