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- Our results show that SAC is well suited for the screening and selection of protein-protein interactions, as exemplified by the rapid generation of specific antibodies against single or multiple antigens.
- SAC selects for diversity rather than affinity. None of the 39 selected scFvs that were sequenced were identical. The avidity effect exploited by SAC allows isolation of low affinity interactions and thus makes maximum use of the diversity encoded in the starting libraries.
- Because of its two-replicon format, pre-existing phage display libraries can be applied directly to SAC with no need for recloning or alterations to existing protocols.
- SAC requires only very low level expression from an incompletely repressed lac promotor, which typicaly yields 100-1000 molecules/cell. Such modest levels of expression should be attainable for a broad range of proteins.
The SAC strategy of assembling high-avidity interaction complexes sufficiently stable and long-lived to allow capture and selection should be generally applicable to other expression hosts and display technologies including in vitro methods like ribosome display and RNA display, provided co-expression of individual receptor-ligand pairs is appropriately compartmentalized.
SAC is a novel, potentially general strategy for the screening and selection of protein-protein interactions based on the assembly and capture of long-lived non-covalent interaction complexes.
As shown, SAC in conjunction with array screening has great potential for the fast generation of a diverse spectrum of antibodies against multiple antigens. We foresee many more applications of the technology, including the screening and selection of generic protein-protein interactions in particular among extracellular or ER proteins, the screening of small molecular compound libraries and the co-evolution of protein-protein interaction surfaces.
R.M.T. deWildt, I.M. Tomlinson, J.L. Ong and P. Holliger. (2002)
Isolation of receptor-ligand pairs by capture of long-lived multivalent interaction complexes on phage.
Proc. Natl. Acad. Sci. USA, 99, 8530-8535.
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