Peptide libraries
The invention relates to a method for altering the conformational diversity of a first repertoire of polypeptide ligands, comprising a plurality of polypeptides comprising at least two reactive groups separated by a loop sequence covalently linked to a molecular scaffold which forms covalent bonds with said reactive groups, to produce a second repertoire of polypeptide ligands, comprising assembling said second repertoire from the polypeptides and structural scaffold of said first repertoire, incorporating one of the following alterations: (a) altering at least one reactive group; or (b) altering the nature of the molecular scaffold; or (c) altering the bond between at least one reactive group and the molecular scaffold; or (d) any combination of (a), (b) or (c).
1. A method for altering the conformation of a first polypeptide ligand or group of polypeptide ligands, wherein each polypeptide ligand comprises at least three reactive groups each separated by a loop sequence and wherein the at least three reactive groups are linked to a molecular scaffold which forms covalent bonds with the at least three reactive groups, to produce a second polypeptide ligand or group of polypeptide ligands, comprising assembling the second ligand or group of ligands from the polypeptide(s) and molecular scaffold of the first ligand or group of ligands by altering at least one of the three reactive groups to cysteine, selenocysteine or methionine.
2. A method according to claim 1 , comprising a further step of incorporating one of: (a) altering a nature of the molecular scaffold that forms covalent bonds with the at least three reactive groups; (b) altering a bond between at least one of the at least three reactive groups; or (c) any combination of (a) or (b).
3. A method according to claim 1 , wherein the reactive groups are cysteine or methionine.
4. A method according to claim 2 , wherein the further step comprises altering a nature of the molecular scaffold that forms covalent bonds with the at least three reactive groups and the altered molecular scaffold is asymmetric.
5. A method according to claim 4 , wherein the molecular scaffold comprises three or more scaffold reactive groups which are capable of forming covalent bonds with the at least three reactive groups on the polypeptides, and wherein two or more reactive groups of the at least three reactive groups are not identical.
6. A method according to claim 5 , wherein one of said scaffold reactive groups is orthogonal.
7. A method according to claim 1 , wherein the scaffold reactive groups are selected from benzylic halides, α-halocarboxylic acids and acryloyl moieties.
8. A method according to claim 1 , wherein at least one bond between the molecular scaffold and the polypeptide are chemically modified after assembly of the polypeptide and the molecular scaffold.
9. A method according to claim 8 , wherein at least one thio-ether linkage between the polypeptide and the molecular scaffold is oxidised to form a sulphoxide or a sulphone.
10. A method according to claim 1 , wherein the first group of polypeptide variants is a first repertoire of polypeptide variants.
11. A method according to claim 1 , wherein the second group of polypeptide variants is a second repertoire of polypeptide variants.