Recombinant Antibody Network

RAN是一个合作的国际三中心伙伴关系，其统一目标是生成定制的 亲和捕获试剂用于人转录因子（TF）。RAN的五年目标是建立一个 强大而有效的管道，将允许可靠地选择高质量的重组抗体（rAB） 这些试剂在质量上是上级的，并且比那些由 杂交瘤技术。生成一套可再生的、经过验证的和标准化的抗体试剂将 使蛋白质功能的研究前所未有，并将加速与生命相关的所有领域的研究 以理工科为重RAN将由一个抗体工程师领导小组指导， 几十年来的合作记录。每个中心都有很高的吞吐量 HTP技术（HTP）平台，具有生产高质量重组抗体（rAB）的成熟能力。 RAN中心的分布模型提供了在要工作的中心之间分布各个任务 以独立的方式，但最终通过输入/输出管道连接在一起，这些管道反馈到所有 网络中的操作。管道的核心部分已经过严格验证， 正如我们的初步结果所显示的那样。我们将建立高通量验证 评估抗体在细胞中从亲和力到活性的性能的程序和ChIP分析。RAN的 HTP管道将使用已建立和优化的自动化噬菌体展示产生rAB，以>85%的TF 抗原由我们的合作者提供，用于总计多达1000个独特的TF。我们将使用HTP验证 每个转录因子产生至少两个rAB的方法，用于在以下方面的高性能：结合亲和力（Kd < 50 nM）、免疫沉淀（IP）和免疫荧光（IF）。我们还将奋进直接验证， 在预算允许的情况下，许多rAB用于染色质免疫沉淀（ChIP）。我们将分发序列 通过多种分销渠道以多种灵活的形式提供信息和开源试剂。的 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。

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.