Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica ›› 2008, Vol. 30 ›› Issue (6): 690-695.doi: 10.3881/j.issn.1000-503X.2008.06.014

• Original Articles • Previous Articles     Next Articles

Construction and Expression of Anti-tumor Necrosis Factor Related Apoptosis-inducing Ligand Receptor Death Receptor 5 Chimeric Antibody in Eukaryotic Cells

CHEN Feng1,2;GUO Ya-bin1;LIU Shi-lian1;ZHENG De-xian1;LIU Yan-xin1   

  1. 1National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005, China 2Laboratory of Molecular Oncology, Peking University School of Oncology, Beijing Cancer Hospital & Institute, Beijing 100142, China
  • Received:2008-04-17 Revised:1900-01-01 Online:2008-12-30 Published:2008-12-30
  • Contact: LIU Yan-xin

Abstract: ABSTRACT:Objective To construct the human/mouse chimeric antibody of a functional anti-death receptor 5 (DR5) antibody. Methods The viable region of light chain (VL) and viable region of heavy chain (VH) genes of anti-DR5 antibody were amplified and cloned into the light- and heavy-chain expression vectors respectively, then the recombinant plasmids were co-transfected into dihydrofolate reductase- Chinese hamster ovary cell (CHO-dhfr-) for expression. The positive clone was screened by the two selective genes (neo and dhfr). The humanization and specificity of chimeric antibody was identified by ELISA and Western bloting, and the tumoricidal activity of the expressed chimeric antibody was detected by tetrazolium salt phenazine methosulfate assay. Results The expression vectors stably expressed chimeric antibody in CHO-dhfr-. In the cell supernatant of the F4’clone, the human IgG heavy constant region and light constant region were identified. Moreover, the secreted chimeric antibody retained the binding capacity to the antigen (DR5) and decreased the cell viability of Jurkat and HCT116 cells to 73.15% and 77.30% in vitro respectively. Conclusion The human/mouse anti-DR5 chimeric antibody has been successfully expressed in eukaryotic cells and shows tumoricidal activity, which establishes a foundation for the future research of humanized antibody medicine.

Key words: anti-deatd receptor 5 cdimeric antibody, cdinese damster ovary cell, duman icc licdt cdain, duman icc deavy cdain