A novel pre-defined CDR library for selection of affinity reagents

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 DESCRIPTION (provided by applicant): The most commonly used method for generating antibodies is through immunization of animals. However, this method is generally low-throughput, expensive, time-consuming, and the antibodies generated are not always renewable. Recombinant antibodies (rAb), like single chain variable fragments (scFv), have many attractive attributes compared to polyclonal antisera and monoclonal antibodies derived from hybridomas. They are renewable through overexpression in the appropriate heterologous host, they are easily stored and transferred as DNA, and they can be genetically engineered as fusions to various enzymes, fluorescent proteins, and epitope tags. While a number of approaches for generating recombinant affinity reagents exist, the cost and throughput of current technologies represent significant roadblocks to the development of a comprehensive and broadly available resource of renewable affinity reagents. We believe that improvements to both gene synthesis technologies and the increased affordability of high-throughput DNA sequencing can be leveraged to create antibody discovery pipelines based on synthetic biology that rival animal immune systems. In this proposal, we will build a high-throughput pipeline for recombinant antibody development in as few as 21 days. The proposed platform takes advantage of pre-designed diversity, next-generation sequence analysis, and advanced molecular biology techniques to enable the rapid identification of specific antibodies. Although our screening platform is being developed with single-chain variable fragment antibodies (scFvs), the technology is applicable to both Fab and yeast display libraries. High-throughput conversion of the scFvs to full immunoglobulin G (IgG) will be integrated within the pipeline so that the antibodies can be directly validated in the desired final format. In genomics, it was thought that the $1000 genome would be the inflection point at which whole genome sequencing would become commonplace. But even at $10,000 per genome, researcher uptake was phenomenal and new ways of using the NextGen sequencers, like ChIP Seq, RNA-seq, etc. were invented. In a similar fashion, we believe that at $5000 and 4 weeks, researchers will begin to develop new applications where the cost of producing antibodies is no longer a relevant factor and speed becomes everything. The ready availability of low cost, high quality affinity reagents will potentially accelerate all aspects of basic science research, provide diagnostics for disease biomarkers, and serve as a proof of concept for therapy. Like the $1000 genome, we think antibody identification and production can eventually go to under $100 and less than 2 weeks. At that point, whole proteome analyses for many different organisms and disease states will be possible. And new methods will arise.