军事医学2012，Vol.36Issue(8)：608-612,625,6.
基于CHO细胞定点整合技术的新型抗体呈现系统的构建
Construction of a new antibody display system based on CHO cell site-specific integration technology
何阳阳 1刘敏 2李春 3张畅 4孙志伟 1刘志刚 2王双 1庞晓斌1
作者信息
- 1. 军事医学科学院生物工程研究所,北京,100071
- 2. 河南大学药学院,开封,475000
- 3. 中国农业大学生命科学学院,北京,100193
- 4. 解放军208医院药剂科,长春,130062
- 折叠
摘要
Abstract
Objective To create a high throughput screening platform for a new antibody display system based on CHO cell site-specific integration technology, on which high affinity anti-VEGF-antibody mutants could be obtained by cell screening. Methods A general plasmid expression vector that could integrate the minibody's gene into CHO cell site-spe-cifically was designed and constructed. To evaluate the display system, three anti-VEGF antibodies named A6,A63 and A657 with known affinity were displayed on CHO cell surface respectively. The efficacy was evaluated through Western blot, Biacore 3000 system and flow cytometry analysis. A light chain antibody library against VEGF was constructed and displayed on CHO cell surface,and specific antibodies were selected by FACS. Results Anti-VEGF antibodies and the library were displayed on CHO cell surface successfully. The relative affinity of minibody on CHO cell surface analyzed by FACS was consistent with the affinity of IgG detected by Biacore 3000 system. Three anti-VEGF-antibody mutants with high affinity were selected through the new antibody display system. Conlusion The new antibody display system based on CHO cell site-specific integration technology can provide new technology support for the antibody's affinity maturation and other molecular mutations.关键词
抗体库/CHO细胞/展示系统/高通量筛选/抗VEGF特异性抗体Key words
antibody library/CHO cells/ display system/high throughput screening/anti-VEGF specific antibody分类
医药卫生引用本文复制引用
何阳阳,刘敏,李春,张畅,孙志伟,刘志刚,王双,庞晓斌..基于CHO细胞定点整合技术的新型抗体呈现系统的构建[J].军事医学,2012,36(8):608-612,625,6.
