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[Objective] The aim of the study was to obtain monoclonal antibody against chicken CD8 molecule. [Method] A fragment of chicken CD8 alpha 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-CD8 alpha was transformed into E. coli, and 39 kDa His-CD8 alpha 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 CD8 alpha 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 the CD8 alpha expressed in prokaryotic system as immunogen.
[Objective] The aim of the study was to obtain a monoclonal antibody against chicken CD8 molecule. [Method] A fragment of chicken CD8 alpha 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 The 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 supernatant was detected by ELISA . A hybridoma cell strain, which could stably excrete antibody against CD8 alpha protein, was obtained and named C11. The ELISA titer of cell supernatant was higher than 1: 640. [Conclusion] A hybridoma cell strain has b een established using the CD8 alpha expressed in prokaryotic system as immunogen.