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学科主题: 基础医学
题名:
Development of single-domain recombinant antibodies to reverse transcriptase domain of human hTERT
作者: Huang, MY; Zhang, B; Wang, JM; Mei, F; Hou, L
刊名: HYBRIDOMA AND HYBRIDOMICS
发表日期: 2004-08-01
卷: 23, 期:4, 页:244-249
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemical Research Methods ; Biotechnology & Applied Microbiology ; Immunology
研究领域[WOS]: Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Immunology
关键词[WOS]: TUMOR-CELLS ; TELOMERASE ACTIVITY ; PHAGE DISPLAY ; MESOTHELIN ; EXPRESSION ; BINDING ; GENE
英文摘要:

This paper describes the development of single-domain recombinant antibodies against human telomerase core protein. A His-tagged hTERT spanning main reverse-transcriptase domain of hTERT was purified from host E. coli and used to immunize BALB/c mice. The VHs (heavy chain variable region genes) were amplified by PCR from total RNA of splenocytes and further induced random mutagenesis by DNA shuffling to enrich the repertoire of VH library. All VHs were cloned into phagemid vectors and displayed to generate 4x10(10) phage libraries. The candidates carrying VH domains against hTERT were primarily screened through three times of panning procedure on His-tagged hTERT coated microplates, and specific antibodies were further selected by West-Western blot. Two clones, designated as a3 and b8, were confirmed to interact with the target in the solid-phase assay. DNA sequencing proved their mouse VH origin. The purified single-domain antibody of b8 could not only recognize native hTERT, but also neutralize human telomerase activity on inhibitory assay and b8 showed the stronger suppressive efficacy compared with a3. The data demonstrated that the developed single-domain recombinant antibodies were hTERT-specific with high potential of binding and activity inhibition.

语种: 英语
WOS记录号: WOS:000223323300006
Citation statistics:
内容类型: 期刊论文
版本: 出版稿
URI标识: http://ir.bjmu.edu.cn/handle/400002259/50784
Appears in Collections:基础医学院_病理学系_期刊论文

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作者单位: Peking Univ, Hlth Sci Ctr, Dept Pathol, Beijing 100083, Peoples R China

Recommended Citation:
Huang, MY,Zhang, B,Wang, JM,et al. Development of single-domain recombinant antibodies to reverse transcriptase domain of human hTERT[J]. HYBRIDOMA AND HYBRIDOMICS,2004,23(4):244-249.
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