Identification of A Monoclonal Antibody against Chicken CD8 Alpha Chain  

作　　者：ZHU Jing-wen YU Wei-yi 

机构地区：[1]Key Laboratory for Zoonoses in Anhui Province, Anhui Agricultural University, Hefei 230036, China

出　　处：《Animal Husbandry and Feed Science》2011年第4期22-23,38,共3页动物与饲料科学（英文版）

基　　金：supported by the grants of the National Natural Science Foundation of China (30671537)

摘　　要：[ Objective] The aim of the study was to obtain monoclonal antibody against chicken CD8 molecule. [ Method] A fragment of chicken CD8 a/pha gene was amplified by PCR with a pair of designed primers. Then two recombinant plasmids containing the amplified fragment were constructed. After prokaryotic expression and purification, the obtained recombinant protein was used to immunize Balb/c mice. Finally, the spleen cells were fused with myeloma cells （SP2/0）, and antibody titer of culture supematant was detected by ELISA. [ Result] A 510-bp gene fragment was amplified by PCR. The recombinant plasmid pET-32a-CD8 alpha was transformed into E. coli, and 39 kDa His-CD8 alpha fusion protein was induced to expression. After subcloning, the culture supernetant was detected by ELISA. A hybridoma cell strain, which could stably excrete antibody against CD8 alpha protein, was obtained and named Cll. The ELISA titer of cell supematant was higher than 1 ： 640. [ Conclusion] A hybridoma cell strain has been established using the CD8 alpha expressed in prokaryoUc system as immunogen.[ Objective] The aim of the study was to obtain monoclonal antibody against chicken CD8 molecule. [ Method] A fragment of chicken CD8 a/pha gene was amplified by PCR with a pair of designed primers. Then two recombinant plasmids containing the amplified fragment were constructed. After prokaryotic expression and purification, the obtained recombinant protein was used to immunize Balb/c mice. Finally, the spleen cells were fused with myeloma cells （SP2/0）, and antibody titer of culture supematant was detected by ELISA. [ Result] A 510-bp gene fragment was amplified by PCR. The recombinant plasmid pET-32a-CD8 alpha was transformed into E. coli, and 39 kDa His-CD8 alpha fusion protein was induced to expression. After subcloning, the culture supernetant was detected by ELISA. A hybridoma cell strain, which could stably excrete antibody against CD8 alpha protein, was obtained and named Cll. The ELISA titer of cell supematant was higher than 1 ： 640. [ Conclusion] A hybridoma cell strain has been established using the CD8 alpha expressed in prokaryoUc system as immunogen.

关 键 词：CHICKEN CD8 alpha Prokaryotic expression Monoclonal antibody 

分 类 号：Q456[生物学—生理学] S852.43[农业科学—基础兽医学]