# US 7,608,187 B2 · Manufacture of a chromatography matrix

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- Number: US 7,608,187 B2
- Document ID: US7608187B2
- Status: Granted
- Kind: B2
- Application number: 12/093,229
- Filed: 2006-11-16
- Published: 2008-10-09
- Granted: 2009-10-27
- Earliest claimed: 2005-11-21
- Assignees: GE Healthcare Bio-Sciences AB
- Inventors: Andreas Axen; Gunnar Glad; Henrik Ihre; Anders Larsson; Nils Norrman
- Web page: https://patentgrep.com/patent/US7608187B2

## CPC

- B01J 20/289: bonded via a spacer
- 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)}
- B01J 20/265: {modified or post-treated polymers (polymer carriers or substrates subjected to further impregnating or coating B01J20/3208)}
- B01J 20/267: {Cross-linked polymers}
- B01J 20/285: based on polymers
- B01J 20/3242: {Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group}
- B01J 20/3244: {Non-macromolecular compounds}
- B01J 39/26: Cation exchangers for chromatographic processes
- B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS

## Abstract

The present invention relates to method of manufacturing a sulphonate-functionalized (S-functionalized) cation exchanger, which method comprises reacting olefinic groups with bisulphite in the presence of at least one amine oxide, such as N-methyl morpholine oxide (NMO). The olefinic groups of the carrier may be allyl groups; which may be provided on extenders such as polyhydroxyfunctional polymers. In one embodiment, the carrier is made of agarose with improved flow pressure properties.

## Claims

### Claim 1

A method of manufacturing a suiphonate-functionalized (S-functionalized) cation exchanger, which method comprises reacting olefinic groups with bisulphite in the presence of at least one amine oxide.

### Claim 2 (depends on 1)

The method of claim 1,wherein the amine oxide is N-methyl morpholine oxide (NMO).

### Claim 3 (depends on 1)

The method of claim 1, wherein the bisulphite is provided by adding sodium bisulphite to a reaction slurry comprising carrier in water.

### Claim 4 (depends on 1)

The method of claim 1, wherein the olefinic groups are allyl groups.

### Claim 5 (depends on 1)

The method of claim 1, wherein the olefinic groups reacted with the bisulphite are provided at the surface of a carrier.

### Claim 6 (depends on 5)

The method of claim 5, wherein the olefinic groups have been coupled to the carrier via extenders.

### Claim 7 (depends on 1)

The method of claim 1, wherein the olefinic groups reacted with the bisulphite are provided on extenders.

### Claim 8 (depends on 7)

The method of claim 7, wherein the extenders are coupled to a carrier after the reaction between olefinic groups and bisulphite.

### Claim 9 (depends on 7)

The method of claim 7, wherein the extenders comprise at least one polyhydroxyfunctional polymer, such as dextran.

### Claim 10 (depends on 5)

The method of claim 5, wherein said carrier is comprised of cross-linked polysaccharide polymers.

### Claim 11 (depends on 10)

The method of claim 10, wherein the cross-linked carrier is provided by:
(a) substituting part of the hydroxyl groups of a polysaccharide in aqueous solution;
(b) providing a carrier by gelling the polysaccharide solution; and
(c) cross-linking the gel obtained by reacting hydroxyl groups of the polysaccharide.

### Claim 12 (depends on 11)

The method of claim 11, wherein the substitution is provided by adding a bifunctional cross-linking agent having one active site and one inactive site to a polysaccharide solution and allowing hydroxyl groups of the polysaccharide to react with the active site of the cross-linking agent; and the inactive site of the cross-linking agent is activated subsequent to the gelling of step (b); which activated site is then reacted with hydroxyl groups of the polysaccharide gel to cross-link the gel.

### Claim 13 (depends on 10)

The method of claim 10, wherein the polysaccharide is selected from the group consisting of agarose and dextran.

### Claim 14 (depends on 13)

The method of claim 13, wherein the polysaccharide is agarose.

### Claim 15 (depends on 10)

The method of claim 10, wherein the carrier is porous.

### Claim 16 (depends on 10)

The method of claim 10, wherein the carrier is comprised of essentially spherical particles.

### Claim 17 (depends on 10)

The method of claim 10, wherein the carrier is a membrane.