Combinatorial libraries of monomer domains
Methods for identifying discrete monomer domains and immuno-domains with a desired property are provided. Methods for generating multimers from two or more selected discrete monomer domains are also provided, along with methods for identifying multimers possessing a desired property. Presentation systems are also provided which present the discrete monomer and/or immuno-domains, selected monomer and/or immuno-domains, multimers and/or selected multimers to allow their selection. Compositions, libraries and cells that express one or more library member, along with kits and integrated systems, are also included in the present invention.
1 . A method for identifying a multimer that binds to a target molecule, the method comprising,
providing a library of monomer domains;
screening the library of monomer domains for affinity to a first target molecule;
identifying at least one monomer domain that bind to at least one target molecule;
linking the identified monomer domains to form a library of multimers, each multimer comprising at least two monomer domains;
screening the library of multimers for the ability to bind to the first target molecule; and
identifying a multimer that binds to the first target molecule.
2 . A polypeptide comprising the multimer selected in claim 1 .
3 . A polynucleotide encoding the multimer selected in claim 1 .
4 . A library of multimers as formed in claim 1 .
5 . A method for identifying a multimer that binds to at least one target molecule, the method comprising:
providing a library of multimers, wherein each multimer comprises at least two monomer domains and each monomer domain exhibits a binding specificity for a target molecule; and
screening the library of multimers for target molecule-binding multimers.
6 . A library of multimers, wherein
each multimer comprises at least two monomer domains connected by a linker; and
each monomer domain exhibits a binding specificity for a target molecule.
7 . A polypeptide comprising at least two monomer domains separated by a heterologous linker, wherein each monomer domain specifically binds to a target molecule.
8 . A method for generating a library of chimeric monomer domains derived from human proteins, said method comprising:
providing loop sequences corresponding to at least one loop from each of at least two different naturally occurring variants of a human protein, wherein the loop sequences are polynucleotide or polypeptide sequences;
covalently combining loop sequences to generate a library of at least two different chimeric sequences, wherein each chimeric sequence encodes a chimeric monomer domain having at least two loops.
9 . A method for identifying an LDL-receptor class A monomer domain that binds to a target molecule, the method comprising,
providing a library of LDL-receptor class A monomer domains;
screening the library of LDL-receptor class A monomer domains for affinity to a target molecule; and
identifying an LDL-receptor class A monomer domain that binds to the target molecule.
10 . A method for identifying a multimer that binds to a target molecule, the method comprising:
providing a library of monomer domains;
screening the library of monomer domains for affinity to a first target molecule;
identifying at least one monomer domain that bind to at least one target molecule;
linking the identified at least one monomer domain to each member of the library of monomer domains to form a library of multimers;
screening the library of multimers for the ability to bind to the first target molecule with an increased affinity; and
identifying a multimer that binds to the first target molecule with an increased affinity.