Polypeptides having a functional domain of interest and methods of identifying and using same
AssigneesThe University of North Carolina at Chapel Hill, Cytogen Corporation
InventorsAndrew B. Sparks, Noah Hoffman, Brian K. Kay, Dana M. Fowlkes, Stephen J. McConnell
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Abstract
Novel polypeptides having functional domains of interest are described, along with DNA sequences that encode the same. A method of identifying these polypeptides by means of a sequence-independent (that is, independent of the primary sequence of the polypeptide sought), recognition unit-based functional screen is also disclosed. Various applications of the method and of the polypeptides identified are described, including their use in assay kits for drug discovery, modification, and refinement.
Claims9
A method of identifying a polypeptide comprising an SH3 domain of interest, comprising
- (a) contacting a multivalent recognition unit complex with a plurality of polypeptides from a cDNA expression library, in which the recognition units are peptides having in the range of 6 to 60 amino acid residues and which selectively bind an SH3 domain;
- and
- (b) identifying a polypeptide having a selective binding affinity for said recognition unit complex;
- wherein the binding specificity of the recognition units has been decreased by incorporating said recognition unit into said multivalent recognition unit complex.
A method of identifying a polypeptide comprising an SH3 domain of interest, comprising:
- (a) contacting a multivalent recognition unit complex, which complex comprises
- (i) avidin or streptavidin, and
- (ii) biotinylated recognition units,
- with a plurality of polypeptides from a cDNA expression library, in which the recognition units are peptides having in the range of 6 to 60 amino acid residues and which selectively bind an SH3 domain;
- and
- (b) identifying a polypeptide having a selective binding affinity for said recognition unit complex;
- wherein the binding specificity of the recognition units has been decreased by incorporating said recognition units into said multivalent recognition unit complex.
Dependencies are inferred from the claim text.