[Preparation and characterization of a mouse anti-human CD40 mutant monoclonal antibody].
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.
Zheng
Shu-Dan
SD
Institute of Medical Biotechnology, Jiangsu Province, Soochow University, Suzhou 215007, China. sweetyzheng@yeah.net
Ma
Hong-Bing
HB
Gao
Chao
C
Whang
Jia-Min
JM
Sun
Jing
J
Luo
Xian-Fu
XF
Zhang
Xue-Guang
XG
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
D000911:Antibodies, Monoclonal; D019013:CD40 Antigens