This Phase 1 proposal plans to determine the feasibility of a novel, high throughput method for screening phage display libraries using micro-encapsulation and FACS. Because of their small size, individual phage cannot currently be screened using FACS. The microspheres generated by One Cell System's technology retain clonal phage resulting in natural signal amplification, permitting flow cytometric analysis.
The aims of the proposal are to use a commercially available phage library that contains approx. 8 avidin binding peptides to determine the feasibility of the system. Phage infected bacteria will be encapsulated in gel microdrops, and streptavidin labeled with Cy-5 used to detect phage making the binding peptides. The ability to sort the drops containing clonal phage populations binding the labeled streptavidin will be assessed. This process will be repeated twice, and then the enriched phage populations grown out and sequenced to determine if they contain bona fide streptavidin binding consensus motifs.
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