Polymeric matrix for affinity chromatography and immobilization of ligands

• US 4,753,983 A
• Filed: 05/07/1986
• Issued: 06/28/1988
• Est. Priority Date: 05/07/1986
• Status: Expired due to Fees

• Current Assignee: Unisyn Fibertec Corporation
• Original Assignee: Bioprobe International Incorporated
• Inventors: Ngo, That T.
• Primary Examiner(s): Phillips, Delbert R.
• Assistant Examiner(s): Nutter, Nathan M.
• Application Number:
  US06/860,603
• Time in Patent Office: 783 Days
• Field of Search: 525/54.11, 525/54.1, 526/238.1, 435/178, 435/179, 435/180, 435/181, 527/300, 530/417, 530/811, 530/812, 530/816
• US Class Current: 525/54.1
• CPC Class Codes: B01J 20/261 obtained by reactions only involving carbon to carbon unsaturated bonds macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds per se C08F
• CPC Class Codes: B01J 20/262 obtained otherwise than by reactions only involving carbon to carbon unsaturated bonds, e.g. obtained by polycondensation macromolecular compounds obtained otherwise than by reactions only involving unsaturated carbon-to-carbon bonds per se C08G
• CPC Class Codes: B01J 20/265 modified or post-treated polymers polymer carriers or substrates subjected to further impregnating or coating B01J20/3208
• CPC Class Codes: B01J 20/267 Cross-linked polymers
• CPC Class Codes: B01J 20/28047 Gels
• CPC Class Codes: B01J 20/289 bonded via a spacer
• CPC Class Codes: B01J 20/291 Gel sorbents
• CPC Class Codes: B01J 20/321 consisting of a polymer obtained by reactions involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3212 consisting of a polymer obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3219 involving a particular spacer or linking group, e.g. for attaching an active group
• CPC Class Codes: B01J 20/3248 the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such
• CPC Class Codes: B01J 20/3255 comprising a cyclic structure containing at least one of the heteroatoms nitrogen, oxygen or sulfur, e.g. heterocyclic or heteroaromatic structures
• CPC Class Codes: B01J 20/3272 Polymers obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/328 Polymers on the carrier being further modified
• CPC Class Codes: B01J 2220/4825 Polysaccharides or cellulose materials, e.g. starch, chitin, sawdust, wood, straw, cotton
• CPC Class Codes: B01J 2220/4831 having been subjected to further processing, e.g. paper, cellulose pulp
• CPC Class Codes: B01J 2220/54 Sorbents specially adapted for analytical or investigative chromatography
• CPC Class Codes: C08F 8/30 Introducing nitrogen atoms or nitrogen-containing groups
• CPC Class Codes: Y10S 530/811 Peptides or proteins is immobilized on, or in, an inorganic carrier
• CPC Class Codes: Y10S 530/812 Peptides or proteins is immobilized on, or in, an organic carrier
• CPC Class Codes: Y10S 530/816 : Attached to the carrier via a bridging agent

Abstract

A matrix useful for immobilizing organic ligands, such as biologically active materials, is prepared by reacting a hydroxyl group containing polymer with polyethyleneimine.

Claims

1. A process for preparing an affinity chromatographic matrix comprising the steps of:

reacting a polymeric substance containing at least one hydroxyl group with 2-fluoro-1-methylpyridinium toluene-4-sulfonate to form an activated polymer wherein at least some of the hydroxyl groups of said polymer have been converted to 1-methyl-2-pyridoxy groups; and

reacting said 1-methyl-2-pyridoxy substituted polymer with polyethyleneimine to form an affinity chromatographic matrix in which at least some of the 1-methyl-2-pyridoxy groups have been replaced by said polyethyleneimine.

2. A process according to claim 1 wherein said polymeric substance is a polysaccharide.

3. A process according to claim 2 wherein said polysaccharide is a dextran.

4. A process according to claim 2 wherein said polysaccharide is a cross-linked dextran.

5. A process according to claim 2 wherein said polysaccharide is an agarose.

6. A process according to claim 2 wherein said polysaccharide is a cross-linked agarose.

7. A process according to claim 2 wherein said polysaccharide is cellulose.

8. A process according to claim 1 wherein said polymeric substance is in the form of a gel.

9. A process according to claim 1 wherein said polymeric substance is a polymer of N-acryloyl-2-amino-2-hydroxymethyl-1,3-propanediol.

10. A process according to claim 1 wherein said polymeric substance is a polyvinyl alcohol.

11. A process according to claim 1 wherein said polymeric substance is paper.

12. A process according to claim 1 wherein said polymeric substance is reacted with said 2-fluoro-1-methylpyridinium toluene-4-sulfonate in aqueous solution.

13. A process according to claim 1 wherein said polymeric substance is reacted with said 2-fluoro-1-methylpyridinium toluene-4-sulfonate in a polar organic solvent.

19. An affinity chromatographic matrix prepared according to the process of claim 1.

14. A process for preparing an affinity chromatographic matrix comprising the steps of:

reacting a polymeric substance containing at least one hydroxyl group with 2-fluoro-1-methylpyridinium toluene-4-sulfonate to form an activated polymer wherein at least some of the hydroxyl groups of said polymer have been converted to 1-methyl-2-pyridoxy groups;

reacting said 1-methyl-2-pyridoxy substituted polymer with polyethyleneimine to form a polyethyleneimine substituted polymer in which at least some of the 1-methyl-2-pyridoxy groups of said activated polymer have been replaced by said polyethyleneimine; and

reacting said polyethyleneimine substituted polymer with an electrophilic ligand capable of reacting with an amino group of said polyethyleneimine-substituted polymer.

15. A process according to claim 14 wherein said electrophilic ligand is a aralkyl carboxylic acid.

16. A process according to claim 14 wherein said electrophilic ligand is ethylene sulfide.

17. A process according to claim 14 wherein said electrophilic ligand is ethylene sulfide and the product of reaction of said polyethyleneimine substituted polymer with ethylene sulfide is reacted with 2,2-dithiopyridine.

18. A process according to claim 14 wherein said electrophilic ligand is a carboxylic acid anhydride.

20. An affinity chromatographic matrix prepared according to the process of claim 14.

21. An affinity chromatographic matrix prepared according to the process of claim 15.

22. An affinity chromatographic matrix prepared according to the process of claim 16.

23. An affinity chromatographic matrix prepared according to the process of claim 17.

24. An affinity chromatographic matrix prepared according to the process of claim 18.