Activated solid support and method
View Patent ↗Disclosed is a method for activating a solid support material with epoxy groups and for immobilizing ligands thereon, utilizing phase transfer catalytic conditions. The method permits the introduction of epoxy groups and specific nucleophilic ligands on the support material with a high level of substitution. Furthermore, the invention provides a general method for immobilizing a ligand for use in a wide variety of chromatographic separation procedures such as ion exchange chromatography, hydrophobic interaction chromatography (HIC), reverse phase chromatography (RPC), or affinity chromatography.
1. A method for preparing a separation matrix, said method comprising reacting together:
a) a matrix having the formula:
M-A-H
wherein:
M is a solid support;
A is an atom or group selected from —O—, —S—and —NR 1 —, where R 1 is selected from H and C 1 -C 4 alkyl; and
b) a compound of formula:
X—(CH 2 ) r —P,
wherein:
X is the group:
wherein R 2 is H; where R 3 is selected from H and C 1 -C 4 alkyl;
P is the group:
where L is a linking group and is a branched or unbranched chain of covalently linked atoms having from 1-20 linear atoms selected from the group consisting of carbon, oxygen and sulphur atoms; and r is 0 or 1;
wherein said method is performed in the presence of a phase transfer catalyst.
2. The method of claim 1 , wherein L is selected from:
where n is an integer from 1 to 6 and m is an integer from 1 to 4.
3. The method of claim 1 , wherein L is the group:
—CH 2 —O—(CH 2 ) n —O—CH 2 — and n is 2, 3 or 4.
4. The method of claim 1 , wherein A is —O—.
5. The method of claim 1 , wherein M is a support particle or bead.
6. The method of claim 1 , wherein M is a porous solid.
7. The method of claim 1 , wherein M is a non-porous solid.
8. The method of claim 1 , wherein M is a cross-linked carbohydrate material.
9. The method of claim 8 , wherein M is selected from the group consisting of agarose, agar, cellulose, dextran, chitosan, konjac, carrageenan, gellan and alginate.
10. The method of claim 9 , wherein M is agarose.
11. The method of claim 1 , wherein M is a synthetic polymer.
12. The method of claim 11 , wherein M is formed from the group consisting of polystyrene, polypropylene, polyvinyltoluene, polyacrylamide, polyacrylonitrile, polycarbonate.
13. The method of claim 1 , wherein M is selected from glass or silica.
14. The method of claim 1 , wherein the phase transfer catalyst is selected from the group consisting of quaternary ammonium salts, phosphonium salts and crown ethers.
15. The method of claim 1 , wherein the phase transfer catalyst is a tetra C 1 -C 8 alkylammonium salt.
16. The method of claim 15 , wherein the phase transfer catalyst is tetrabutylammonium hydrogen sulphate.
17. The method of claim 1 , wherein the reaction is performed under basic conditions at a pH range of between 9 and 13.