Recombinant Antibody Network

重组抗体网络

• 批准号: 8221472
• 负责人: ANTHONY A KOSSIAKOFF
• 金额: $ 323.34万
• 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/8221472
• 关键词: Affinity; Animals; Antibodies; Antibody Affinity; Back; Binding; Biological Sciences; Biomedical Research; Cells; Characteristics; Generations; Goals; Human; Hybridomas; Immunization; Immunofluorescence Immunologic; Immunoprecipitation; Individual; International; Modeling; Monoclonal Antibodies; Output; Performance; Phage Display; Procedures; Proteome; Reagent; Recombinant Antibody; Research; Technology; Time; Validation; Western Blotting; Work; antibody engineering; base; chromatin immunoprecipitation; cost; feeding; high throughput technology; open source; operation; protein function; transcription factor

The RAN is a collaborative international tri-center partnership having the unified goal of generating customized affinity capture reagents to human transcription factors (TFs). 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 hybridoma 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 (HTP) 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 HTP pipelines will generate rABs using established and optimized automated phage display to >85% of the TF antigens provided by our collaborators for up to a total of 1000 unique TFs. We will use HTP validation procedures to produce at least two rABs per transcription factor for high performance in: binding affinity (Kd < 50 nM), immunoprecipitation (IP) and immunofluorescence (IF). We will also endeavor to validate directly as many rABs for use in chromatin immunoprecipitation (ChIP) as the budget allows. We will distribute sequence information and open source reagents in several flexible 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.

RAN是一个协作的国际三场伙伴关系，具有生成定制的统一目标 亲和力捕获对人转录因子（TFS）的试剂。跑步的五年目标是建立一个 强大而有效的管道将允许可靠选择高质量重组抗体（RAB） 质量优越且生产更便宜，生产的试剂比由 杂交瘤技术。生成可再生，经过验证和标准化的抗体试剂集 启用对蛋白质功能的前所未有的研究，并将加速与生命有关的所有领域的研究 科学。跑步将由领先的抗体工程师团体指导，他们的轨道已建立 数十年来共同努力的记录。每个中心都有很高的吞吐量 技术（HTP）平台具有生成高质量重组抗体（RABS）的可靠功能。 RAN中心的分布模型规定，在要工作的中心之间分配单个任务 以独立的方式进行，但最终由输入/输出管道绑在一起 网络中的操作。管道的核心部分已经过严格验证，并且具有出色的 我们的初步结果所证明的能力。我们将建立高通量验证 评估从亲和力到细胞活性和芯片分析活动的抗体性能的程序。跑了 HTP管道将使用已建立和优化的自动噬菌体显示到> 85％的TF生成RABS 我们的合作者提供了多达1000个独特的TFS的抗原。我们将使用HTP验证 在：结合亲和力（KD < 50 nm），免疫沉淀（IP）和免疫荧光（如果）。我们还将努力直接验证 预算允许的许多RAB用于染色质免疫沉淀（芯片）。我们将分发序列 通过多种分发渠道以几种灵活的格式提供信息和开源试剂。这 该项目的影响将是加速速率并降低转录因子研究成本（TFS） 特别是蛋白质组。