Method of preparing affinity ligands
View Patent ↗The present invention is a method of preparing affinity ligands, which method comprises to provide a cyclic scaffold comprising a thiol group, a carbonyl group and an amine group; to derivatize the amine group of said scaffold with an electrophile that carries either a functionality capable of affinity interaction or a functionality capable of a second interaction with a target molecule; and to open up the derivatized scaffold and add an amine that carries either a functionality capable of a second interaction or a functionality capable of affinity interaction with a target molecule. The method provides provide affinity ligands, which are capable of affinity interaction as well as a second kind of interaction.
1. A method of preparing at least one affinity ligand, which method comprises
(a) providing a cyclic scaffold comprising a thiol group, a carbonyl group and an amine group;
(b) derivatising the amine group of said scaffold with an electrophile that carries either a functionality capable of affinity interaction or a functionality capable of a second interaction with a target molecule; and
(c) opening up the cyclic structure of the derivatised scaffold and adding to the carbonyl carbon an amine that carries either a functionality capable of a second interaction or a functionality capable of affinity interaction with a target molecule;
to result in a ligand capable of affinity interaction with one site of a target molecule and a second functionality capable of interacting with another site of the same target molecule.
2. The method of claim 1 , wherein the functionality that is capable of providing affinity interaction is a known affinity ligand, and the method results in a ligand of modified characteristics as compared to said known affinity ligand.
3. The method of claim 1 , wherein step (b) is performed with an electrophilic carboxyl acid carrying a functionality capable of affinity interaction and step (c) adds an amine that carries a second functionality.
4. The method of claim 1 , wherein step (b) is performed with an electrophile carrying a second functionality and step (c) adds an amine that carries a functionality capable of affinity interaction.
5. The method of claim 1 , wherein the scaffold is defined by the general formula (I)
wherein A, B and X irrespective of each other are carbon atoms or heteroatoms, m is any integer between 0 and 4, and the functional group N is a nitrogen, which either replaces one X or is attached to any one of A, B and X.
6. The method of claim 5 , wherein in formula (I), A, B and X are carbon atoms.
7. The method of claim 5 , wherein in formula (I), m is 1 and the scaffold is homocysteine thiolactone.
8. The method of claim 1 , wherein the electrophile is selected from the group consisting of compounds containing carbon-carbon double bonds conjugated to Y, wherein Y is a halogen, a mesylate group, a tosylate group, an acid or an activated acid.
9. The method of claim 1 , wherein the functionality capable of affinity interaction is one member of an affinity pair.
10. The method of claim 1 , wherein the functionality is capable of interacting with a target molecule by another interaction than affinity.
11. The method of claim 1 , wherein steps (a) and (b) have been performed earlier to provide a ready-derivatised scaffold.
12. The method of claim 1 , wherein the product obtained from step (c) is immobilised via its thiol group to a base matrix in order to produce an affinity separation medium comprising a plurality of affinity ligands.
13. The method of claim 12 , wherein the thiol group is coupled to allyl groups of the base matrix.