title: [Preparation and characterization of a mouse anti-human CD40 mutant monoclonal antibody].
abstract: OBJECTIVE
  To prepare and characterize a mouse anti-human CD40 mutant monoclonal mAb.
  
  
  METHODS
  Female BALB/c mice of 6-8 weeks old were immunized with CD40 mutant transfectant (L929-CD40mu) as immunogen. The spleen B cells of the mice were fused with Sp2/0 myeloma cells.The hybridoma cells were screened with CD40 mutant transfectant (L929-CD40mu) by FCM. Fast-strip analysis was performed to identify Ig subclass of this mAb. The epitope recognized by this mAb was detected by Bio-5C11 competitive assay. Western blot technique was adopted to identify the mAb. The proliferation of tumor cells in vitro was analyzed by MTT assay and apoptosis of tumor cells in vitro was analyzed by PI-annexin V assay.
  
  
  RESULTS
  One hybridoma cell line named 10C5 was obtained, which had the property of secreting anti-human CD40 mutant monoclonal antibody continuously and steadily. This mAb specifically recognized human CD40 mutant molecule and induced the apoptosis of tumor cells in vitro.
  
  
  CONCLUSIONS
  One hybridoma cell line which can secret a mouse anti-human CD40 mutant mAb has been prepared successfully. This mAb can inhibit the growth of tumor cells expressing CD40 mutant and induce their apoptosis in vitro.
authors.affiliation: Institute of Medical Biotechnology, Jiangsu Province, Soochow University, Suzhou 215007, China. sweetyzheng@yeah.net
authors.forename: Shu-Dan
authors.initials: SD
authors.lastname: Zheng
authors.forename: Hong-Bing
authors.initials: HB
authors.lastname: Ma
authors.forename: Chao
authors.initials: C
authors.lastname: Gao
authors.forename: Jia-Min
authors.initials: JM
authors.lastname: Whang
authors.forename: Jing
authors.initials: J
authors.lastname: Sun
authors.forename: Xian-Fu
authors.initials: XF
authors.lastname: Luo
authors.forename: Xue-Guang
authors.initials: XG
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology*; D017209:Apoptosis / Q000276:immunology; D019013:CD40 Antigens / Q000276:immunology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005260:Female; D005786:Gene Expression Regulation / Q000276:immunology; D006801:Humans; D006825:Hybridomas / Q000166:cytology; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation
chemical_list: D000911:Antibodies, Monoclonal; D019013:CD40 Antigens