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 是一个国际三中心合作伙伴关系，其统一目标是生成定制的 人类转录因子 (TF) 的亲和捕获试剂。 RAN 的五年目标是建立一个 强大而高效的管道，可以可靠地选择高质量的重组抗体 (rAB) 与通过化学方法生产的试剂相比，其质量更优，生产成本更低且生产速度更快 杂交瘤技术。生成一套可再生、经过验证和标准化的抗体试剂将 实现对蛋白质功能的前所未有的研究，并将加速与生命相关的所有领域的研究 科学。 RAN 将由拥有既定轨道的领先抗体工程师团队领导 数十年的合作记录。每个中心都将拥有高吞吐量 技术 (HTP) 平台具有生成高质量重组抗体 (rAB) 的经过验证的能力。 RAN 中心的分布式模型允许在要工作的中心之间分配各个任务 以独立的方式开启，但最终通过输入/输出管道连接在一起，反馈到所有 网络中的操作。该管道的核心部件经过严格验证，具有卓越的性能 我们的初步结果表明了这一能力。我们将建立高通量验证 评估抗体性能（从亲和力到细胞活性）和 ChIP 分析的程序。 RAN 的 HTP 管道将使用已建立和优化的自动化噬菌体展示来生成 rAB，以达到 > 85% 的 TF 我们的合作者为总共 1000 个独特的 TF 提供了抗原。我们将使用 HTP 验证 每个转录因子至少产生两个 rAB 的程序，以实现以下方面的高性能： 结合亲和力 (Kd < 50 nM)、免疫沉淀 (IP) 和免疫荧光 (IF)。我们还将努力直接验证 在预算允许的情况下，许多 rAB 可用于染色质免疫沉淀 (ChIP)。我们将分配序列 通过多种分销渠道以多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快转录因子（TF）研究的速度并降低其成本 特别是蛋白质组。