Construction and expression of human mouse chimeric antibody directed against TNF-α

Objective:Construction and expression of TNF α human mouse chimeric antibody CZ8.Methods:The light and heavey chain variable genes were inserted into the chimeric antibody expression vectors and transfected into SP 2/0 or CHO cells.The expression of anti TNF α human mouse chimeric antibody was tested by ELISA、RT PCR and Western blot.The in vitro neutralization assay was used to test the biological activity of chimeric antibody.Results:Anti TNF α human mouse chimeric antibody was expressed by transfected CHO cells with a single vector that contained both light chain and heavy chain genes.ELISA analysis showed the TNF α chimeric antibody was specific for TNF α.The transcription of human mouse chimeric light and heavy chain mRNAs were proved by RT PCR.Protein expression of both κ and H chain was further verified by Western blot.The in vitro neutralization assay demonstrated that the chimeric antibody could neutralize the cyctoxicity of TNF α.Conclusion:Human mouse chimeric antibody CZ8 was successfully expressed in eukaryotic cells.