Development And Characterization Of Monoclonal Antibodies To VP22 Of Marek's Disease Virus

1 Prokaryotic Expression of VP22 of Marek's disease virus CVI988/Rispens StrainThe gene fragments coding for the C-terminus (amino acids 94-243,VP22C), the deleted N-terminus (aa 19-243,VP22H), the wild type one(VP22), and the terminus between aa 207-249(VP22T) of CVI988/Rispens VP22, were respectively subcloned into the prokaryotic expression vector pGEX-6p-1,and transformed into the competent cells of BL21 (DE3) E. coli. After 3 h incubation, expression of the truncated VP22 protein was induced by 0.05mmoL IPTG. After brief sonication and centrifugation, the supernatant of the cell lysate was separated by 12% SDS-PAGE and the band of interest was excised and minced for mouse immunization. Balb/c mice were immunized peritoneally with 50μg of the E. coli - expressed truncated VP22 protein in minced gel slice and boosted every 10 d for 3 times. Seven days after the final immunization, the serum was collected for immunofluorescence assay (IFA) and western-blot (WB) analysis on CEF cells of the second passage infected with MDV-1 strains CVI988, GA, and RB1B. IFA and Western-blot results showed that the product of VP22was a good immugen.2 Development and its characteristic of Monoclonal Antibody to VP22 of Marek's disease virusFive strains of hybridoma cells secrecting monoclonal antibody (mAb) against VP22were obtained by fusion between SP2/0 myeloma cells and spleen cells of balb/c mice immunized with the fusion protein VP22C-GST. Specificity assay showed that these mABs only reacted with CEFs infected with MDV-1 in IFA and had no cross-reaction with NDV,REV,ADV and IBDV. The IFA titer of these mAbs reacted to...