Solid-phase immobilization of proteins and peptides
View Patent ↗Disclosed herein are methods for immobilizing a peptide or protein on a solid support. The method generally includes the following steps: (a) providing one or more templates attached to a solid support, wherein the one or more templates include (i) an RNA encoding a peptide and (ii) a peptide acceptor-linker linked to the RNA; and (b) subjecting the one or more templates to conditions that support translation and attachment of said peptide to said peptide acceptor, thereby synthesizing the one or more peptides on the solid support. Also disclosed herein are solid supports having at least one RNA-protein fusion component immobilized thereon, methods for generating protein arrays, and methods for screening molecules using these arrays.
1. A solid support having at least one RNA-protein fusion component immobilized thereon, wherein said RNA-protein fusion component comprises a protein covalently bonded at its C-terminus to a peptide acceptor, wherein said peptide acceptor is joined through a linker to the RNA which encodes the protein, wherein the linker comprises a non-nucleotide moiety, said RNA-protein fusion component being attached to the solid support by the interaction of said linker and said solid support.
2. The solid support of claim 1 , wherein said linker comprises a biotin component and the solid support comprises a steptavidin component and said RNA-protein fusion component is attached to said solid support via binding of said biotin component to said streptavidin component.
3. The solid support of claim 1 , wherein said linker comprises a sequence that is complementary to a portion of said RNA and is hybridized to said portion of said RNA.
4. The solid support of claim 3 , wherein said sequence is a PNA sequence.
5. The solid support of claim 3 wherein said linker further comprises a psoralen moiety that is cross-linked to said RNA.
6. The solid support of claim 1 , wherein said support comprises a plurality of immobilized proteins that form an array.
7. A method for screening for a molecule in a population of molecules for its ability to interact with a protein, said method comprising the steps of:
(a) providing the solid support of claim 1 ;
(b) contacting a population of molecules with said solid support; and
(c) detecting the interaction of a molecule in said population with an immobilized protein of said solid support, thereby screening for a molecule in a population of molecules for its ability to interact with a protein.
8. A method for screening for a protein in a plurality of proteins for its ability to interact with a component of a sample, said method comprising the steps of:
(a) providing the solid support of claim 1 ;
(b) contacting said sample with said solid support; and
(c) detecting an interaction of a component of said sample with an immobilized protein of said solid support, thereby screening for a protein in a plurality of proteins for its ability to interact with a component of a sample.