Title: Prokaryotic expression and development of monoclonal antibody against glycoprotein 3(GP3) of PRRSV CH-1a strain
Abstract: The ORF3 gene of PRRSV strain CH1a was truncated and cloned into a prokaryotic expression vector pGEX-6p-1,and a fusion-expressed protein rtGP3 of 40 000 was obtained in E.coli.The protein rtGP3 showed a strong reaction to the PRRSV-positive sera in Western blotting.BALB/c mice were immunized intraperitoneally with purified rtGP3 protein.Murine myeloma cells were fused with the splenocytes of the immunized mice after the third immunization.An indirect ELISA coated with rtGP3 and GST was used to screen hybridomas for production of specific antibody in hybridoma culture fluid.Five hybridomas clone which produced McAbs steadily was obtained,named 31A2,31G11,32B12,32D4 and 32H7.The McAbs showed strong reactivity in IFA on the PRRSV infected Marc-145 cells.The McAbs belong to IgG1 isotype,and their light chain are κ chain,which was isotyped by ELISA using a commercial kit.The McAbs against PRRSV GP3 protein could be used for further analysis of the structure and function of PRRSV.
Publication Year: 2007
Publication Date: 2007-01-01
Language: en
Type: article
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