Universal peptide-binding scaffolds and protein chips
AssigneeThe Board of Trustees of the University of Illinois
InventorsHuimin Zhao, Olga Esteban
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Abstract
The present invention provides a universal peptide-binding scaffold. This scaffold is used to bind a target. The target can be a peptide or peptides of interest (for example, peptides associated with a disease state) or can represent the entire proteome. The target can be either protein fragments prepared by enzymatic digestion of the entire proteome or N- or C-terminal short sequences exposed by chemical denaturation of the entire proteome (unfolded proteins). The universal peptide-binding scaffold can be tailored to specifically bind a target using the methods described herein.
Claims14
A universal peptide or protein binding scaffold comprising:
- a library of mutants of a peptide or protein binding scaffold of MHC class II DR1 peptide binding domains having an affinity for a ligand between 10−6 and 10−9 molar and having a point mutation L11H in the β1 domain.
A protein chip comprising:
- a substrate and mutants of a peptide or protein binding scaffold of MHC class II DR1 peptide binding domains having a point mutation L11H in the β1 domain bound to the substrate, wherein the peptide has an affinity for a ligand between 10−6 and 10−9 molar.
A method of selecting proteins or peptides that bind to a peptide binding scaffold comprising:
- preparing a library of mutants of a peptide binding domain of MHC class II peptide binding domains having a point mutation L11H in the β1 domain;
- contacting said library with labeled peptides or proteins;
- and selecting those mutants that bind to labeled peptides or proteins with a desired affinity.
Dependencies are inferred from the claim text.