[Preparation and immunological characterization of monoclonal antibodies against VP22 carboxyl terminus of Marek's disease virus serotype 1].
OBJECTIVE
To prepare and characterize monoclonal antibodies (mAb) against VP22 carboxyl terminus of CVI988/Rispens strain of Marek's disease virus serotype 1.


METHODS
Carboxyl terminus of CVI988 VP22 (94aa-243aa) was expressed in prokaryotic system. mAbs against VP22 were prepared by hybridoma technology from BALB/c mice immunized with the fusion protein GST-VP22C and characterized by ELISA, indirect fluorescence assay (IFA) and Western blot.


RESULTS
Two hybridoma cell lines stably secreting mAb against VP22C were obtained and designated as 3F7 and 4E4. mAb 3F7 could react with VP22 expressed in all the plaques, while mAb 4E4 stained all the cells nuclei in MDV-infected CEF cells. It was also found that 3F7 could react with VP22 expressed in Sf9 cells and denatured VP22 by Western blot analysis. In addition, it was further showed that the epitope of mAb 3F7 was located within the domain between 94aa and 193aa, the predicted site of protein transduction domain of VP22.


CONCLUSIONS
The preparation of the mAb is very important to further research in protein transduction domain of MDV-1 VP22.
Song
Cui-Ping
CP
Key Lab of Jiangsu Preventive Veterinary Medicine, College of Veterinary Medcine, Yangzhou University, Yangzhou, China.
Chen
Hong-Jun
HJ
Qin
Ai-Jian
AJ
Zhang
Chen-Fei
CF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D002460:Cell Line; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D008381:Herpesvirus 2, Gallid / Q000276:immunology* / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins