title: [Construction and stable expression of anti-human tumor necrosis factor-related apoptosis-inducing ligand receptor 2 chimeric antibody in CHO cells].
abstract: OBJECTIVE
  To establish an human-mouse chimeric antibody against tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor 2 (death receptor 5, DR5) in an eukaryotic cell line and analyse its tumoricidal activity.
  
  
  METHODS
  The cDNAs encoding for the variable regions of heavy chain (V(H);) and light chain (V(L);) of AD5-10 were amplified by PCR and inserted into the human IgG heavy and light chain containing expression vector RpCI-neo, respectively. The recombinant plasmids were co-transfected into HEK293 and/or CHO cells. The production of anti-DR5 human-mouse chimeric antibody (hmAD5-10) and the antibody affinity for DR5 were identified by ELISA and Western blot assay. The tumoricidal activity of hmAD5-10 was demonstrated by MTS assay. The stable expression cells were selected and cultured in serum-free medium.
  
  
  RESULTS
  Two stable CHO cells CHO-A5 and CHO-B11 with the chimeric antibody hmAD5-10 expression were established, in which the production of hmAD5-10 were reached at (0.36±0.11) mg/L and (0.16±0.01) mg/L, respectively. The hmAD5-10 secreted from the cells can well bind with DR5 and kill the cultured leukemia SVT35 cells by apoptosis remarkably.
  
  
  CONCLUSIONS
  The human-mouse chimeric antibody hmAD5-10 was successfully expressed in the eukaryotic cells and resulted tumor cell death by apoptosis. This study lays a fundamental basis for the potential application of the recombinant chimeric antibody in cancer therapy.
authors.affiliation: National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS, Beijing 100005, China. safe1017@sina.com
authors.forename: Fu-jia
authors.initials: FJ
authors.lastname: Lv
authors.forename: Juan
authors.initials: J
authors.lastname: Shi
authors.forename: Ya-xi
authors.initials: YX
authors.lastname: Zhang
authors.forename: Shi-lian
authors.initials: SL
authors.lastname: Liu
authors.forename: Yan-xin
authors.initials: YX
authors.lastname: Liu
authors.forename: De-xian
authors.initials: DX
authors.lastname: Zheng
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics* / Q000627:therapeutic use; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D051379:Mice; D009369:Neoplasms / Q000188:drug therapy; D018124:Receptors, Tumor Necrosis Factor / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000627:therapeutic use
chemical_list: D000906:Antibodies; D018124:Receptors, Tumor Necrosis Factor; D011993:Recombinant Fusion Proteins; C114109:TNFRSF21 protein, human