Identification of a monoclonal antibody against chicken CD8 alpha chain.

[Objective] The aim of the study was to obtain monoclonal antibody against chicken CD8 molecule.[Method] A fragment of chicken CD8α 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 supernatant was detected by ELISA.[Result] A 510-bp gene fragment was amplified by PCR.The recombinant plasmid pET-32a-CD8α was transformed into E.coli,and 39 kDa His-CD8α fusion protein was induced to expression.After subcloning,the culture supernatant was detected by ELISA.A hybridoma cell strain,which could stably excrete antibody against CD8α protein,was obtained and named C11.The ELISA titer of cell supernatant was higher than 1:640.[Conclusion] A hybridoma cell strain has been established using CD8α expressed in prokaryotic system as immunogen.