Recombinant Antibody Network

重组抗体网络

• 批准号: 8725993
• 负责人: ANTHONY A KOSSIAKOFF
• 金额: --
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-26 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8725993
• 关键词: Affinity; Animals; Antibodies; Antibody Affinity; Antibody Formation; Antigens; Architecture; Area; Arizona; Back; Binding; Biological Assay; Biological Sciences; Biology; Biomedical Research; Budgets; Cell Line; Cells; Characteristics; Chicago; Data; Feedback; Generations; Goals; Human; Hybridomas; Immunization; Immunofluorescence Immunologic; Immunoglobulin G; Immunoprecipitation; Individual; Intellectual Property; International; Internet; Length; Link; Modeling; Monoclonal Antibodies; Output; Performance; Phage Display; Plasmids; Procedures; Process; Production; Protein Binding; Proteins; Proteome; Protocols documentation; Reagent; Recombinant Antibody; Recombinant Proteins; Research; Research Personnel; San Francisco; Speed; Technology; Time; Universities; Validation; Vendor; Western Blotting; Work; antibody engineering; base; chromatin immunoprecipitation; cost; design; expression vector; feeding; flexibility; high throughput technology; improved; member; open source; operation; process optimization; protein function; repository; restraint; success; transcription factor

DESCRIPTION (provided by applicant): The RAN is a collaborative international tri-center partnership having the unified goal of generating customized affinity capture reagents to all the human transcription factors. The five-year goal of the RAN is to establish a robust and efficient pipeline that will allow for reliable selection of high quality recombinant antibody (rAB) reagents that are superior in quality and are much cheaper and quicker to produce than those generated by hybridism technology. Generation of a renewable, validated and standardized set of antibody reagents would enable unprecedented studies of protein function and would accelerate research in all fields related to life sciences. The RAN will be directed by a leading group of antibody engineers who have an established track record of working collaboratively together for decades. Each of the centers will have a high throughput technology platform with the proven capabilities to generate high quality recombinant antibodies (rABs). The distributive model of the RAN centers provides for distributing individual Tasks among centers to be worked on in an Independent manner, but ultimately tied together by input/output conduits that feed back into all operations in the network. The core pieces of the pipeline have been rigorously validated and have exceptional capacity as demonstrated in our Preliminary Results. We will have established high-throughput validation procedures that assess antibody performance from affinity to activity in cells and ChIP analysis. The RAN's high throughput (HTP) pipelines will generate rABs in multiple formats using established and optimized automated phage display to 1500 human TFs provided by our collaborators and others. We will use HTP validation procedures to produce at least two rABs per transcription factor for high performance in: binding (KD <50 nM), Western blotting, immunoprecipitation (IP), immunofluorescence (IF) and chromatin immunoprecipitation (ChIP). We will distribute sequence information and open source reagents in multiple formats through multiple distribution channels. The impact of this project will be to accelerate the rate and reduce the cost of research in transcription factors (TFs) specifically and the proteome in general.

描述（由申请人提供）：RAN是一个协作的国际三场伙伴关系，其统一目标是将定制的亲和力捕获试剂与所有人类转录因子产生。 RAN的五年目标是建立可靠的高质量重组抗体（RAB）试剂的强大而有效的管道 质量优越，比杂种技术生成的产品便宜得多，生产更快。生成可再生，经过验证和标准化的抗体试剂集将实现前所未有的蛋白质功能研究，并在与生命科学有关的所有领域中加速研究。跑步将由领先的抗体工程师团体指导，他们在数十年中共同努力合作的良好记录。每个中心都将具有高吞吐技术平台，并具有生成高质量重组抗体（RABS）的良好功能。 RAN中心的分布模型规定，以独立的方式在要处理的中心之间分配各个任务，但最终由输入/输出管道捆绑在一起，这些输入/输出管道将其反馈到网络中的所有操作中。管道的核心部分已经过严格验证，并且具有出色的能力，如我们的初步结果所示。我们将建立高通量验证程序，以评估从亲和力到细胞活性和芯片分析的抗体性能。运行的高吞吐量（HTP）管道将使用已建立和优化的自动化噬菌体显示器以多种格式生成RAB，我们的合作者和其他人提供了1500个人类TF。我们将使用HTP验证程序每次转录至少产生两个RAB，以在：结合（KD <50 nm），蛋白质印迹，免疫沉淀（IP），免疫荧光（IF）和染色质免疫沉淀（CHIP）中。我们将通过多种分布通道以多种格式分发序列信息和开源试剂。该项目的影响将是加速速率并降低转录因子（TFS）和一般蛋白质组的研究成本。