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.
A major limitation in biomedical research is the lack of high quality antibody affinity reagents. The goal of this research is to generate a new class of recombinant antibody reagents with superior characteristics that will effectively replace traditional antibodies based on animal immunization. They will also be produced quicker, cost less and will be renewable.
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