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学科主题基础医学
Development of single-domain recombinant antibodies to reverse transcriptase domain of human hTERT
Huang, MY; Zhang, B; Wang, JM; Mei, F; Hou, L
刊名HYBRIDOMA AND HYBRIDOMICS
2004-08-01
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
引用统计
被引频次:1[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
版本出版稿
条目标识符http://ir.bjmu.edu.cn/handle/400002259/50784
专题北京大学基础医学院_病理学系
北京大学基础医学院
北京大学公共卫生学院_北京大学儿童青少年卫生研究所
北京大学第一临床医学院_儿科
作者单位Peking Univ, Hlth Sci Ctr, Dept Pathol, Beijing 100083, Peoples R China
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GB/T 7714
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.
APA Huang, MY,Zhang, B,Wang, JM,Mei, F,&Hou, L.(2004).Development of single-domain recombinant antibodies to reverse transcriptase domain of human hTERT.HYBRIDOMA AND HYBRIDOMICS,23(4),244-249.
MLA Huang, MY,et al."Development of single-domain recombinant antibodies to reverse transcriptase domain of human hTERT".HYBRIDOMA AND HYBRIDOMICS 23.4(2004):244-249.
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