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