Modification of polypeptides
The invention provides a method for conjugating a peptide displayed on a genetic display system to a molecular scaffold performed on an ion exchange resin.
1. A method for conjugating a peptide displayed on a phage display system to a molecular scaffold, comprising the steps of:
(a) combining polypeptides displayed on a phage display system with a purification resin such that the phage display system is bound to the resin and treating the bound phage display system with a reducing agent;
(b) exposing the bound phage display system to the molecular scaffold;
(c) removing unreacted molecular scaffold from the bound phage display system; and
(d) eluting the phage display system from the purification resin.
2. A method according to claim 1 , wherein the phage is a wild-type phage.
3. A method according to claim 1 , wherein step (a) is followed by a washing step before addition of the molecular scaffold.
4. A method according to claim 3 , wherein the phage display system is washed in a dilute solution of reducing agent.
5. A method according to claim 3 , wherein the wash solution further comprises a chelating agent.
6. A method according to claim 1 , wherein the reducing agent is TCEP.
7. A method according to claim 1 , wherein the scaffold is TBMB.
8. A method according to claim 1 , wherein the molecular scaffold is added in the presence of aqueous acetonitrile.
9. A method according to claim 1 , wherein the resin is an anion exchange resin.
10. A method according to claim 1 , wherein the resin is magnetic.
11. A method according claim 1 , wherein one or both of steps (a) and (b) is performed at room temperature (25° C.).
12. A method according to claim 1 , wherein step (a) is performed for 20 minutes.
13. A method according to claim 1 , wherein step (b) is performed for 10 minutes.