Preparation and characterization of a mouse anti-human CD40 mutant monoclonal antibody
10.3321/j.issn:1007-8738.2009.10.014
- VernacularTitle:一株识别肿瘤细胞上CD40突变体分子的单克隆抗体的研制及其生物学功能研究
- Author:
Shudan ZHENG
;
Hongbing MA
;
Chao GAO
;
Jiamin WANG
;
Jing SUN
;
Xianfu LUO
;
Xueguang ZHANG
- Publication Type:Journal Article
- Keywords:
CD40;
hybridoma;
monoclonal antibody;
mutant;
tumor cell
- From:
Journal of Cellular and Molecular Immunology
2009;25(10):910-913
- CountryChina
- Language:Chinese
-
Abstract:
AIM: 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. Faststrip 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. CONCLUSION: 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.
