Patent ID: 9644201
Date: 2017-05-09
CPC Classifications: A61K,C07K,C12N,C40B,G01N

Claim:
1. A method for preparing one or more peptide ligands, comprising the steps of (a) displaying one or more polypeptides in a phage display system, wherein the polypeptide comprises SEQ ID No. 1, SEQ ID No. 12, SEQ ID No. 13, SEQ ID No. 14, SEQ ID No. 15, SEQ ID No. 16, or SEQ ID No. 17; (b) providing tris-(bromomethyl)benzene (TBMB); (c) attaching TBMB to said polypeptide by formation of three discrete covalent bonds between said TBMB and said polypeptide via cysteine amino acid residues of SEQ ID No. 1, SEQ ID No. 12, SEQ ID No. 13, SEQ ID No. 14, SEQ ID No. 15, SEQ ID No. 16, or SEQ ID No. 17, and form at least one loop which comprises a sequence of two or more amino acids subtended between two of said cysteine amino acid residues, thus forming the one or more peptide ligand; (d) exposing the peptide ligands to one or more proteases, wherein exposure to the protease is carried out in the presence of a reducing agent; and (e) screening the peptide ligands for binding against a target, and selecting the ligands which bind to the target, wherein the peptide ligands selected for binding to the target are resistant to protease cleavage.