Development And Preliminary Application Of Hybridoma Cell Strains Respectively Secreting Monoclonal Antibodies Against PRV And PRRSV

A porcine pseudorabies virus(PRV)strain and a porcine reproductive and respire tory syndrome virus(PRRSV)strain were isolated and identified from some pig farms in Henan Province.Balb/c mice were immunized with purified PRV and PRRSV as the mix antigen,.Hybridoma cell strains were obtained that secreted the monoclonal antibodies separately against the two antigens above:two strains against PRV named 7F5,7G3 and three strains against PRRSV named 2D5,2D9,1F7.At the same time, Hybridoma cell strain which was named 6E11 was obtained from the Balb/c mice immunized against PRV by the traditional fuse method.Hybridoma cell strain which was named 3E2 was obtained from the immuned Balb/c mice against PRRSV by the traditional fuse method.The anti-PRV monoclonal antibodies have no cross-reaction with other viruses.The anti-PRRSV monoclonal antibodies have also no crossreaction with other viruses which indicated they were sepecific.Titers of monoclonal antibodies against PRV in cell cultures and mouse ascites were over 1:1600 and 1;51200 respectively.Titers of hybridoma cells against PRRSV in hybridoma cultures and mouse ascites were over 1:3200 and 1:51200 respectively.These hybridoma cells strains were stable to secrete MaAbs.Mouse ascites were purifid and the protein concentrations were 52.95 mg/mL for MaAb against PRV and 56.25 mg/mLfor MaAb against PRRSV respectively.The developments of the hybridoma cell strains lay a foundation for the kit of PRV and PRRSV in further.Compared with the traditional method,this research developed the method of mix immunities,screening respectively.Hybridoma cell fusion method against different antigens which we has developed is a highly effective method.This method can not only save the experimental material and reduce cost,but also save time and manpower,which can greatly improve work efficiency.The work can be completed by one worker now, compared with two or more workers in the past time.It is a vital significance to the hybridlump research and the practical application.Antigen capture enzyme-linked immunosorbent assay(AC ELISA)was developed to detect the PRRSV antigen with the home-made anti-PRRSV monoclonal anti- bodies.This method can detect PRRSV from tissues and cell cultures but it can not differentiate virulent virus from the vaccine virus.Even though,this method still has the importancy to detect the unimmunized swine herds,the vaccine virus or the cell cultures of virulent virus.