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| 1 | 鸡球虫病免疫防治研究进展显示文摘从鸡球虫强毒疫苗、弱毒疫苗、重组抗原苗、单克隆抗体以及单链抗体等方面,阐述了鸡球虫病免疫预防的发展过程及鸡球虫病免疫防治的发展方向。 | 李艳琴 王振海 秦建华 李潭清 | 2008 | 安徽农业科学2008,36,13: | 19 |
| 2 | Screening and evaluation of human single-chain fragment variable antibody against hepatitis B virus surface antigen显示文摘BACKGROUND: Phage display technology has become a vital tool in studies aimed at identifying molecules binding to a specific target. It enables the rapid generation and selection of high affinity, fully human antibody product candidates to essentially any disease target appropriate for antibody therapy. In this study, we prepared the recombinant single-chain fragment variable ( ScFv) antibody to hepatitis B virus surface antigen (HBsAg) by the phage display technology for obtaining a virus-targeting mediator. METHODS: mRNA was isolated from B-lymphocytes from a healthy volunteer and converted into cDNA. The fragment variables of heavy and light chain were amplified separately and assembled into ScFv DNA with a specially constructed DNA linker by polymerase chain reaction. The ScFv DNA was ligated into the phagmid vector pCANT-AB5E and the ligated sample was transformed into competent E. coli TG1. The transformed cells were infected with M13K07 helper phage to form a human recombinant phage antibody library. The volume and recombinant rate of the library were evaluated by bacterial colony count and restriction analysis. After two rounds of panning with HBsAg. the phage clones displaying ScFv of the antibody were selected by enzyme-linked immunosorbant assay ( ELISA) from the enriched phage clones. The antigen binding affinity of the positive clone was detected by competition ELISA. HB2151 E. coli was transfected with the positive phage clone demonstrated by competition ELISA for production of a soluble form of the anti-HBsAg ScFv. ELISA assay was used to detect the antigen binding affinity of the soluble anti-HBsAg ScFv. Finally, the relative molecular mass of soluble anti-HBsAg ScFv was measured by SDS-PAGE. RESULTS: The variable heavy ( VH ) and variable light (VL) and ScFv DNAs were about 340bp, 320bp and 750bp, respectively. The volume of the library was up to 2 × 106 and 8 of 10 random clones were recombinants. Two phage clones could strongly compete with the original HBsAb for binding to HBsAg. Within 2 strong positive phage clones, the soluble anti-HBsAg ScFv from one clone was found to have the binding activity with HBsAg. SDS-PAGE showed that the relative molecular weight of soluble anti-HBsAg ScFv was 32 kDa. CONCLUSION: The anti-HBsAg ScFv successfully produced by phage antibody technology may be useful for broadening the scope of application of the antibody. | Jian-Lin Zhang, Jian-Jin Guo, Zi-Yan Zhang, Yi-Xin Jing, Lin Zhang, Rui Guo, Ping Yan, Niu-Liang Cheng, Bo Niu and Jun Xie Department of Biochemistry and Molecular Biology, Shanxi Medical University ,Taiyuan 030001,China | 2006 | Hepatobiliary & Pancreatic Diseases International2006,5,2: | 8 |
| 3 | 单链抗体在抗肿瘤和抗感染免疫治疗中的应用显示文摘单链抗体(single-chain antibody fragment,scFv)是保持了亲本抗体的抗原识别特异性及亲和活性的最小功能性抗体片段,其良好的可塑性及广泛作用已使之成为研究的热点。scFv相关的研究成果不仅改进了已有的肿瘤和病毒感染诊治方法,还提出了新颖的抗肿瘤、抗感染思路,如靶向肿瘤新生血管及细胞外基质相关蛋白的提出改变了以往仅针对肿瘤细胞的狭隘治疗思维。文章综述了scFv在抗肿瘤和抗感染方面的研究及应用。 | 田锋 彭国平 | 2007 | 医学分子生物学杂志2007,4,5: | 3 |
| 4 | 前列腺癌患者噬菌体抗体库的构建显示文摘【目的】构建前列腺癌患者的噬菌体抗体库,为前列腺癌特异性的人源性抗体Fab片段的筛选及其应用奠定基础。【方法】由前列腺癌患者外周血分离得淋巴细胞,完整提取其总RNA,经RT-PCR及半套式扩增得到免疫球蛋白全部轻链和重链Fd段基因,然后经酶切、纯化、连接等步骤克隆进载体pComb3,并电转化大肠杆菌XL1-Blue,从而构建前列腺癌患者噬菌体抗体库。【结果】成功构建噬菌体抗体Fab片段库,其轻链及重链Fd段与载体DNA的重组率分别为87%、79%,库容为23.2×107,滴度为87×1012mL-1。【结论】本研究成功构建了前列腺癌患者噬菌体抗体库。 | 赵风进 黄健 林天歆 黄海 许可慰 潘秋辉 | 2004 | 中山大学学报(医学科学版)2004,25,4: | 1 |
| 5 | Phage display creates innovative applications to combat hepatitis B virus显示文摘Hepatitis B virus(HBV) has killed countless lives in human history. The invention of HBV vaccines in the 20 th century has reduced significantly the rate of the viral infection. However, currently there is no effective treatment for chronic HBV carriers. Newly emerging vaccine escape mutants and drug resistant strains have complicated the viral eradication program. The entire world is now facing a new threat of HBV and human immunodeficiency virus co-infection. Could phage display provide solutions to these life-threatening problems? This article reviews critically and comprehensively the innovative and potential applications of phage display in the development of vaccines, therapeutic agents, diagnostic reagents, as well as gene and drug delivery systems to combat HBV. The application of phage display in epitope mapping of HBV antigens is also discussed in detail. Although this review mainly focuses on HBV, the innovative applications of phage display could also be extended to other infectious diseases. | Wen Siang Tan Kok Lian Ho | 2014 | World Journal of Gastroenterology2014,20,33: | 1 |