MOLECULAR IMMUNOLOGY

分子免疫学

• 批准号: 3838451
• 负责人: T S PAPAS
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3838451
• 关键词: affinity chromatography; genetic library; genetic manipulation; genetic techniques; immunoglobulin genes; immunoglobulin structure; laboratory mouse; molecular cloning; molecular oncology; monoclonal antibody; oncoproteins; protein engineering; recombinant DNA

In the past year, some advances have been made in mimicking in vivo antibody generation by use of molecular biology approaches. The most significant breakthrough was the development of the phage Fab/Fv display library technology. There is interest in exploring the application of this technology to cancer research and antibody engineering. An anti-ETS2 monoclonal antibody T-7 light chain, U-244 heavy chain, and a presumably heavy chain-related sequence from both T-7 and U-244, have been cloned successfully. A new Fab combinatorial library construction strategy for a more diverse repertoire has been designed. Mouse splenic anti-erg Fab cDNAs are to be cloned and screened by this strategy. The phage Fab display vector has been requested from MRC. An anti-erg Fab display library will be made to verify the efficiency of the system. Some BALB/c mice have been immunized with several clinical tumor samples and their normal controls. Fab display libraries will be made, and those anti-tumor marker clones will be selected by panning or affinity chromatography. For high-affinity Fab, a new approach to generate random mutagenesis to desired domains has been started and this work will continue. An approach to generate a new generation of therapeutic monoclonal antibodies by combining desired specificity associated with antibody variable domains with the defined antibody effector function associated with some monoclonal antibody constant domains has been designed. Several monoclonal antibody cell lines are to be characterized.

在过去的一年里，在体内模拟方面取得了一些进展， 通过使用分子生物学方法产生抗体。 最 噬菌体Fab/Fv展示技术的发展是一个重大突破 图书馆技术 有兴趣探讨应用 这项技术应用于癌症研究和抗体工程。 抗ETS 2 单克隆抗体T-7轻链、U-244重链，以及推测的 已经克隆了来自T-7和U-244重链相关序列 成功地 一种新的Fab组合文库构建策略 设计了更加多样化的曲目。 小鼠脾抗erg Fab 通过该策略克隆和筛选cDNA。 噬菌体Fab 已向MRC请求显示矢量。 一种anti-erg Fab显示器， 并将其应用于图书馆，以验证系统的效率。 一些BALB/c 已经用几种临床肿瘤样品免疫小鼠， 正常对照者 Fab展示文库将被制作，并且那些抗肿瘤抗体将被制备。 标记克隆将通过淘选或亲和层析来选择。 为 一种产生随机突变新方法， 已经开始了所需的领域，这项工作将继续下去。 方法 以产生新一代的治疗性单克隆抗体， 结合与抗体可变结构域相关的所需特异性 与一些相关的定义的抗体效应子功能， 设计了单克隆抗体恒定结构域。 几 将表征单克隆抗体细胞系。