[Preparation of a novel functional anti-human CD40 monoclonal antibody and identification of its bioactivity].
OBJECTIVE
To prepare a novel functional mouse anti-human CD40 monoclonal mAb.


METHODS
Female BALB/c mice of 6-8 weeks old were immunized with CD40 transfectant (L929-CD40) as immunogen. The spleen B cells of the mice were fused with Sp2/0. The hybridoma cells were screened with CD40 transfectant (L929-CD40) 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. The proliferation and cell cycle of tumor cells in vitro were studied by MTT assay and PI staining respectively.


RESULTS
One hybridoma cell line named 2B6 was obtained, which had the property of secreting anti-human CD40 monoclonal antibody continuously and steadily. This mAb specifically recognized human CD40 molecule and promoted the proliferation of tumor cells in vitro.


CONCLUSIONS
One hybridoma cell line which can secret a novel functional mouse anti-human CD40 mAb has been developed successfully. This mAb can specifically recognize human CD40 and influence the growth of tumor cells in vitro.
Tang
Lin
L
Institute of Medical Biotechnology, Key Laboratory of Medicine and Clinical Immunology of Jiangsu Province, Soochow University, Suzhou 215007, China. tl915@163.com
Zheng
Lu
L
Qi
Chun-jian
CJ
Ma
Hong-bing
HB
Zhou
Xuan
X
Zhuang
Yu-Mei
YM
Luo
Xian-fu
XF
Xu
Ying
Y
Zhang
Xue-guang
XG
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology*; D000918:Antibody Specificity / Q000276:immunology*; D001667:Binding, Competitive; D019013:CD40 Antigens / Q000276:immunology*; D002453:Cell Cycle / Q000276:immunology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004306:Dose-Response Relationship, Immunologic; D000939:Epitopes / Q000276:immunology; D005260:Female; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D009369:Neoplasms / Q000473:pathology*; D013997:Time Factors