| 人源性抗乳腺癌Her-2/neu噬菌体单链抗体库构建 |
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| 引用本文： | 李晓莉,姚慧,周登远,焦振山. 人源性抗乳腺癌Her-2/neu噬菌体单链抗体库构建[J]. 生物医学工程与临床, 2010, 14(2): 161-166,F0002. DOI: 10.3969/j.issn.1009-7090.2010.02.020 |
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| 作者姓名： | 李晓莉 姚慧 周登远 焦振山 |
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| 作者单位： | 天津市中医药研究院,附属长征医院,中心实验室,天津,300120 |
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| 基金项目： | （志谢：感谢天津医科大学附属肿瘤医院乳腺科张斌博士从标本采集、实验方法指导等多方面给予的支持与帮助.） |
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| 摘 要： | 目的构建乳腺癌患者抗Her-2/neu全人源性噬菌体单链抗体可变区（ScFv）文库,筛选抗Her-2/neu单链抗体。方法收集40例乳腺癌患者前哨淋巴结（SLN）,提取总RNA,反转录为cDNA作模板,在目前常用的引物基础上重新设计可变区的引物,用PCR扩增全套抗体可变区,构建T载体库。并改造pCANTAB-5E构建了pCANTAB-Linker。将T载体库酶切连接入pCANTAB-Linker构建出ScFv文库。最后构建噬菌体单链抗体库并进行初级筛选。结果成功改良ScFv文库的构建方法。初步得到12株对人乳腺癌组织有高亲和力的Her-2/neu单链抗体。结论改良了ScFv文库的构建方法,可以获得较大库容量的单链抗体库,并从中筛选到人源性抗Her-2/neu单链抗体。
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| 关 键 词： | 乳腺癌 Her-2/neu 单链抗体 |
Construction of a phage single-chain antibody library human-derived anti-breast cancer Her-2/neu |
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| LI Xiao-li,YAO Hui,ZHOU Deng-yuan,JIAO Zhen-shan. Construction of a phage single-chain antibody library human-derived anti-breast cancer Her-2/neu[J]. Biomedical Engineering and Clinical Medicine, 2010, 14(2): 161-166,F0002. DOI: 10.3969/j.issn.1009-7090.2010.02.020 |
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| Authors: | LI Xiao-li YAO Hui ZHOU Deng-yuan JIAO Zhen-shan |
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| Affiliation: | LI Xiao-li,YAO Hui,ZHOU Deng-yuan,JIAO Zhen-shan(Center Laboratory,Tianjin Changzheng Hospital Affiliated Tianjin Institute of Traditional Chinese Medicine,Tianjin 300120,China) |
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| Abstract: | Objective To construct a phage single-chain fragment variable region(ScFv) antibody library of human-derived antibreast cancer Her-2/neu,and screen out anti-Her-2/neu specific single-chain antibody from the library.Methods Total RNA was extracted from 40 sentinel lymph nodes(SLN) of breast cancer patients and reversely transcribed into cDNA.After a group of the primers of variable regions were redesigned based on the current commonly used ones,the PCR was used in amplified a full set of antibody variable re... |
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| Keywords: | Her-2/neu |
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