Recombinant Antibody Network

描述(由申请人提供)：RAN是一个协作性的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产生定制的亲和捕获试剂。RAN的五年目标是建立一个强大和高效的管道，以便能够可靠地选择高质量的重组抗体(RAb)试剂 与杂交技术产生的产品相比，这些产品质量更高，生产成本更低，速度更快。一套可再生的、有效的和标准化的抗体试剂的产生将使蛋白质功能的研究成为可能，并将加快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小组指导，他们有几十年合作的既定记录。每个中心都将拥有一个高通量的技术平台，具有生产高质量重组抗体(RAB)的成熟能力。RAN中心的分布式模型提供了在要以独立方式工作的中心之间分配单个任务，但最终通过反馈到网络中所有操作的输入/输出管道将其捆绑在一起。正如我们的初步结果所表明的那样，管道的核心部件已经过严格的验证，并具有出色的能力。我们将建立高通量验证程序，评估抗体性能，从亲和力到细胞活性和芯片分析。RAN的高吞吐量(HTP)管道将使用我们的合作者和其他人提供的针对1500个人类TF的已建立和优化的自动化噬菌体展示，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生产至少两个RAB，以便在以下方面获得高性能：结合(Kd&lt；50 NM)、Western blotting、免疫沉淀(IP)、免疫荧光(IF)和染色质免疫沉淀(CHIP)。我们将通过多个分发渠道，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)和一般蛋白质组的研究速度并降低成本。

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.