Fibronectin type III domain based scaffold compositions, methods and uses
A protein scaffold based on a consensus sequence of fibronectin type III (FN3) proteins, such as the tenth FN3 repeat from human fibronectin (human Tenascin), including isolated nucleic acids that encode a protein scaffold, vectors, host cells, and methods of making and using thereof have applications in diagnostic and/or therapeutic compositions, methods and devices. In particular, protein scaffold molecules binding to IgG have been identified as useful for diagnostic and/or therapeutic applications.
1. A method of constructing a library of a protein scaffold based on a fibronectin type III (FN3) domain derived from a consensus sequence of an FN3 domain, comprising the steps of:
providing a polypeptide derived from a consensus sequence of an FN3 domain having at least 90% identity to the amino acid sequence of SEQ ID NO:16; and
introducing diversity into copies of the polypeptide to form the protein scaffold library.
2. A method of constructing a library of a protein scaffold based on a fibronectin type III (FN3) domain, comprising the steps of:
providing a polypeptide having the amino acid sequence of SEQ ID NO:16; and
introducing diversity into copies of the polypeptide having the amino acid sequence of SEQ ID NO:16 to form the protein scaffold library.
3. The method of claim 2 , wherein the introducing diversity step comprises mutating at least one loop region selected from the group consisting of residues at or about positions 13-16, 22-28, 38-43, 51-54, 60-64, and 75-81 of SEQ ID NO:16.