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In Vitro Culture for H5N1-Specific Duck T Cells and Detection of Immune Responses Using Intracellular Cytokine Staining Method
Published on: May 30, 2025
[Secretable eukaryotic expression of recombinant chicken Iglambda and preparation of monoclonal antibodies against
Hong Zhang1, Gai-ping Zhang, Jin-gang Zhang
1Academy of Agricultural Sciences in Henan, Zhengzhou 450002, China. angela9922@sina.com
Aim:
To construct the eukaryotic expression vector for chicken Iglambda light chain, to express it on COS7 cells and to prepare the monoclonal antibodies against chicken Iglambda.
Methods:
The cDNA of chicken Iglambda light chain with signal peptide sequence was amplified and then inserted into eukaryotic expression plasmid pcDNA3 after double enzyme cutting. The constructed recombinant vector was transfected into COS7 cells by lipofectamin and the secretable eukaryotic expression of chicken Iglambda light chain was verified by Western blot. The monoclonal antibodies (mAbs) against chicken Iglambda light chain were prepared by immunizing BALB/c mice with 2 x 10(6) chicken B cells and by cell fusion technology.
Results:
The eukaryotic expression vector was successfully constructed. Western blot demonstrated that chicken Iglambda light chain existed in the cultural supernatant. The hybridoma lines secreting anti-Iglambda mAbs were screened by indirect ELISA. The specific reactivity between anti-Iglambda mAbs and recombinant chicken Iglambda light chain was detected by Western blot.
Conclusion:
The secreted recombinant chicken Iglambda light chain and anti-Iglambda mAbs provide a basis for further study of the functions of chicken Iglambda.
