Patent ID: 8685890
Filing Date: 2014-04-01
Classification: A61K,C07K,C12N,C40B,G01N

Abstract:
1. A method for providing a multispecific peptide ligand comprising a polypeptide covalently linked to a molecular scaffold at three or more amino acid residues and capable of binding to two or more separate targets, comprising the steps of: (a) providing a first phage displayed repertoire of polypeptides, each polypeptide comprising two or more reactive groups capable of covalent linkage to tris-(bromomethyl)benzene (TBMB), and at least one loop which comprises a sequence of two or more amino acids subtended between two of said reactive groups; (b) providing a second phage displayed repertoire of polypeptides as described in (a); wherein said first and second phage displayed repertoires are screened for binding to first and second targets to isolate one or more members for use in step (c); wherein said first and second phage displayed repertoires are screened for binding by conjugating said members of said first and second phage displayed repertoire to TBMB, and screening for binding of said first and second phage displayed repertoires against said first or second target; wherein the members of said first and second phage displayed repertoires are conjugated to said TBMB to form at least one loop; wherein said TBMB is present at less than 100 μM during conjugating to said first and second phage displayed repertoires; wherein said first and second phage displayed repertoires of polypeptides are encoded by a first and a second library of nucleic acid molecules; wherein the screening of said first and second phage displayed repertoires is performed using phage display; (c) joining at least one loop of one or more members of the phage encoded first repertoire to at least one loop of one or more members of the phage encoded second repertoire to form at least one polypeptide comprising two loops, and (d) conjugating the composite polypeptide(s) to a molecular scaffold at at least three amino acid positions, thereby forming the multispecific peptide ligand.