The Prepare Of Monoclonal Antibodies Against Protein σB Of Muscovy Duck Reovirus

Muscovy duck reovirus(MDRV)disease is one kind of high incidence,high mortality muscovy duck infective disease.It has caused serious economic losses in the muscovy duck industry since it happened,and became one of the main epidemic diseases which restrict the healthy development of the muscovy duck industry in our country.Muscovy duck reovirus is a type of RNA virus.The particles are spherical or nearly spherical Icosahedron with a 60nm～73nm length in diameter.It is Symmetric three-dimensional,non-capsular,and have visible double-capsid structure.Muscovy duck reovirus is composed by sub-segments of 10 gene fragments,which code for 10 proteins,including 8 proteins for the capsid structural proteins and 2 non-structural proteins,dividing into three groups,λ,μandσ.Among them,the study of proteinσC is the more sophisticated,which is the major structural protein reculting by the immunogenicity.While,it was also found in recent years,non-structural proteinsσB UnitingσC immune can played better recults.It has not been reported about the role of the mechanism of proteinσB in the domestic.In this study,pET30-σB,having His-tag,was transformed into E.coli BL21(DE3)and expressed toσB,which is 45kDa as expected,by Recombinant plasmids recombinanted in laboratory.The His-Bind Purification Kit was used to purify the protein,which was the antigen. The recults of Western blots analysis Muscovy Duck Reovirus positive serum first antibody showed that.A specific reaction occered betweenσB and positive serum of Muscovy duck reovirus,which indicate thatσB has good immunogenicity.Immunizing 8 weeds female mouse with the depuranted and renaturated renaturat protein,the researcher prepared forσB Monoclonal antibodies of Muscovy duck reovirus.Through a series of basic identification,4 sorts of monoclonal hybridoma were confirmed,named A3F,B2B,C3E and D2G.Moreover,the natural structure of the proteins were expressed in Sf9 cells by baculovirus,and the immunohistochemisty recults of the 4 sorts of monoclonal hybridoma showed that they all have reaction with natruel Conformation proteins,which means they are great valuable.Also,the preliminary epitope analysis of the Monoclonal antibodies showed that A3F corresponds to 1～47 amino acids,B2B and C3E corresponds to 1～80 amino acids and D2G corresponds to 48～80 amino acids.