Generation of fully human anti-vascular endothelial growth factor 165 monoclonal antibody with antitumor efficacy from transgenic five-feature mice
10.3321/j.issn:1000-5048.2009.03.016
- VernacularTitle:利用五特征转基因小鼠制备全人源血管内皮生长因子165单克隆抗体
- Author:
Yu LIU
;
Yue GU
;
Dongjie YUN
;
Jianquan CHEN
;
Siguo LIU
;
Aimin ZHANG
- Publication Type:Journal Article
- Keywords:
five-feature mice;
fully human mAb;
antitumor;
VEGF_(165)
- From:
Journal of China Pharmaceutical University
2009;40(3):269-272
- CountryChina
- Language:Chinese
-
Abstract:
Aim: To prepare a fully human anti-VEGF_(165)(vascular endothelial growth factor 165) monoclonal antibody with antitumor activity from five-feature mice which express human immunoglobin loci. Methods: A routine method for the generation of monoclonal antibodies( mAbs) against the human VEGF_(165) was developed. The immunizing effect between five-feature mice and BALB/c was observed and the mAb was purified through MBP IgM affinity chromatography. The effect of mAbs on antitumor was tested ire vitro by T24 cell line. Results: Four hybri-domata secreting mAbs steadily were isolated successfully, and the serum titer of mAb in BALB/c mice was almost 10 times higher than that in five-feature mice. The indirect ELISA method for mAb titer determination was also established. The anti-VEGF_(165) mAb was purified to homogeneity by precipitation with ammonium sulfate followed by the affinity chromatography on MBP IgM purification column. Moreover, both purified human IgM V_2, V_(75) and mouse ascites were characterized by SDS-PAGE and Western blotting. Proliferation of T24 cell line was considerably inhibited by V_2 and V_(75). Conclusion: Five-feature mice could be used to produce fully human monoclonal antibody. The fully human anti-VEGF mAb is potential in the cancer treatment.
