Recombinant Antibody Network

重组抗体网络

• 批准号: 8545885
• 负责人: ANTHONY A KOSSIAKOFF
• 金额: $ 180.73万
• 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 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8545885
• 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是一个国际三中心合作伙伴关系，其统一目标是为所有人类转录因子生成定制的亲和捕获试剂。RAN的五年目标是建立一个强大而有效的管道，以便可靠地选择高质量的重组抗体（rAB）试剂 其在质量上是上级的，并且比通过杂交技术产生的那些更便宜和更快地生产。一套可再生、经过验证和标准化的抗体试剂的产生将使前所未有的蛋白质功能研究成为可能，并将加速与生命科学相关的所有领域的研究。RAN将由一个领先的抗体工程师团队领导，他们在数十年的合作中建立了良好的记录。每个中心都将拥有一个高通量技术平台，具有生产高质量重组抗体（rAB）的能力。RAN中心的分布式模型提供了在中心之间以独立的方式分配各个任务，但是最终通过反馈到网络中的所有操作的输入/输出管道捆绑在一起。管道的核心部分已经过严格验证，并具有卓越的能力，正如我们的初步结果所示。我们将建立高通量验证程序，以评估抗体在细胞和ChIP分析中从亲和力到活性的性能。RAN的高通量（HTP）管道将使用我们的合作者和其他人提供的1500个人类TF的已建立和优化的自动化噬菌体展示来生成多种格式的rAB。我们将使用HTP验证程序为每个转录因子产生至少两个rAB，以在以下方面实现高性能：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）。我们将通过多种分销渠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快转录因子（TF）和蛋白质组研究的速度并降低其成本。