Study On Rapid Preparation Of Avian Influenza Virus H5N1 Antibody

Avian influenza(AI)is a poultry infectious disease that spreads around the world.It has high mortality rate,is highly contagious and can spread across species,seriously affecting the development of the poultry industry and other aquaculture industries as well as human health.The avian influenza virus(AVI)belongs to the influenza A virus.The H5N1 virus belongs to the influenza A virus subtype and is mainly found in poultry.It is a highly pathogenic virus that can cause humans and many animals to become ill.In order to effectively suppress the spread of the disease,effective vaccines have been produced.However,currently used vaccines are mostly inactivated vaccines,and there are still certain problems in terms of production cost,stability of potency,and protection.Hemagglutinin(HA)is an antigenic substance on the surface of avian influenza virus that is responsible for the binding of the virus to the host receptor.Antibody(Ab)is a glycoprotein produced by plasma cells(effector B cells)formed by proliferation and differentiation after B cells receive antigenic stimulation,and is mainly present in body fluids such as serum,and is an important effector molecule that mediates the immunity of a conductor.It exerts immune function after specifically binding to the antigen.Using the specificity of binding of antibodies to antigens,antibodies have been widely used for the detection of viruses,bacteria,and foreign proteins,and can be used as agents or drugs for diagnosing and treating certain diseases in humans and animals.Antibodies are classified into natural antibodies,monoclonal antibodies(mAbs),polyclonal antibodies(polyclonal antibodies),and genetically engineered antibodies.Among them,monoclonal antibodies and polyclonal antibodies are widely used.The specificity of monoclonal antibodies is high,but the preparation procedure is cumbersome,the experiment period is long,and the price is high.Preparation of polyclonal antibodies is mostly performed by subcutaneous injection or intravenous injection of prokaryotic protein to achieve antibody production,but the use of antibody titers in eukaryotic cells remains to be improved.In general,the immunized animals used for the preparation of polyclonal antibodies are mainly mammals and birds,and they need to be selected according to actual requirements such as the use and demand of the antibodies.As a kind of edible microorganism,yeast has the advantages of high safety,strong immune adjuvant effect,and strong resistance to gastrointestinal secretions.It is reported that yeast can present recombinant proteins to animals and cause immune reactions.The laboratory has for the first time demonstrated that yeast can present DNA fragments to mouse intestine dendritic cells(DCs)and induce immune responses in mice.Later,it was demonstrated that yeast can present foreign DNA to animals such as fish and rabbits.Therefore,we used yeast as the antigen presentation vector for this experiment to achieve rapid antibody preparation.In this study,we designed a recombinant yeast that expresses the HA antigen gene of avian influenza H5N1 virus and presents it to BALB/c mice by oral feeding to achieve rapid preparation of H5N1 antibody.First,the eukaryotic expression vector JMB84-CMV-HA containing the H5N1 HA antigen gene was constructed;then,the JMB84-CMV-HA vector was transformed into S.cerevisiae JMY1,and the HA gene was presented orally to the mouse intestinal DCs.After 28 days of immunization,serum HA protein antibodies were detected by Western Blotting.This study provides new ideas for the preparation of antibodies against avian influenza virus.