[1]翟伟,郝长付,高彬,等.尘肺病噬菌体单链抗体库的构建及鉴定*[J].郑州大学学报(医学版),2011,(06):852.
 ZHAI Wei,HAO Changfu,GAO Bin,et al.Construction and identification of human phage singlechain antibody library for pneumoconiosis[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2011,(06):852.
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尘肺病噬菌体单链抗体库的构建及鉴定*
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2011年06期
页码:
852
栏目:
论著
出版日期:
2011-11-20

文章信息/Info

Title:
Construction and identification of human phage singlechain antibody library for pneumoconiosis
作者:
翟伟郝长付高彬姚武#
郑州大学公共卫生学院劳动卫生学教研室 郑州 450001
Author(s):
ZHAI WeiHAO ChangfuGAO BinYAO Wu
Department of Occupational Health,College of Public Health, Zhengzhou University,Zhengzhou 450001
关键词:
尘肺噬菌体单链抗体库单链抗体
Keywords:
pneumoconiosisphage singlechain antibody libraryScFv
分类号:
R135.2
摘要:
目的:构建具有良好多样性的人尘肺病噬菌体单链抗体(ScFv)库。方法:收集25例尘肺病患者外周血标本共50 mL,密度梯度离心法分离淋巴细胞,Trizol法提取总RNA,逆转录后参考文献设计并合成半巢式PCR引物,扩增抗体重链(VH)和轻链(VL)可变区基因,Linker连接VH和VL,继而连接到噬菌体载体pCANTAB5E中,连接产物转入感受态大肠杆菌TG1,超感染辅助噬菌体M13KO7,构建噬菌体ScFv库。结果:扩增出VH和VL基因序列并有效地连接,经转化和超感染,构建了库容为2.4×107 pfu/mL的ScFV库。结论:成功构建人尘肺病噬菌体ScFV库。
Abstract:
Aim: To construct a human pneumoconiosis phage singlechain antibody(ScFv) library with diversity.Methods:Total RNA was extracted from the peripheral blood lymphocytes from the patients with pneumoconiosis, VH and VL genes were amplified by seminested PCR and were assembled to form ScFv by Linker and cloned into phagemid pCANTAB5E, and then transformed into E. coli TG1 to construct a human peumoconiosis phage scFv antibody library.Results:VH gene families and VL gene families were successfully amplified. A large human antibody library containing 2.4×107 pfu/mL clones was created after rescuing the recombinant phagemids from the transformed E.coli TG1 cells.Conclusion: A human ScFv library is successfully constructed finally.

参考文献/References:

[1]Maestrelli P, Guarnieri G. Diagnosis of occupational chronic pulmonary diseases[J]. G Ital Med Lav Ergon,2010,32(4 Suppl):433 [2]Sblattero D,Bradbury A.A definitive set of oligonucleotide primers for amplifying human V regions[J].Immunotechnology,1998,3(4):271 [3]Barboza CE,Winter DH,Seiscento M,et al.Tuberculosis and silicosis:epidemiology,diagnosis and chemoprophylaxis[J].J Bras Pneumol,2008,34(11):959 [4]Ilavska S,Jahnova E,Tulinska J,et al.Immunological monitoring in workers occupationally exposed to asbestos[J].Toxicology,2005,206(2):299 [5]Otusuki T, Maeda M, Murakami S, et al. Immunological effects of silica and asbestos[J].Cell Mol Immunol,2007, 4(4):261 [6]王雪涛.矽肺与免疫[J].职业与健康,2007,23(20):1085 [7]苏建华,郑宏,崔力争.抗人尘细胞单克隆抗体用于低SiO2粉尘所致尘肺的诊断研究[J].中国职业医学,2001,28(4):6 [8]于秀艳,张皓,孙敏莉,等.聚合酶链反应扩增人动脉粥样硬化抗体全套可变区基因[J].上海交通大学学报:医学版,2006,26(11):1260 [9]Wright MJ,Deonarain MP.Phage display of chelating recombinant antibody libraries[J].Mol Immunol,2007,44(11):2860 [10]王晋,罗文新,李利峰,等.多样性人源天然噬菌体抗体库的构建及初步应用[J].细胞与分子免疫学杂志,2006,22(6):786 [11]王琰,王刚,化冰.人噬菌体抗体库中异常重组子的分析[J].细胞与分子免疫学杂志,2002,18(1):63

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备注/Memo

备注/Memo:
*国家自然科学基金资助项目81072278 #通讯作者,男,1962年8月生,博士,教授,研究方向:生产性粉尘与尘肺,Email:yaowu@zzu.edu.cn
更新日期/Last Update: 1900-01-01