# US 9,433,923 B2 · Chromatography media and use thereof

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- Number: US 9,433,923 B2
- Document ID: US9433923B2
- Status: Granted
- Kind: B2
- Application number: 14/407,892
- Filed: 2013-06-11
- Published: 2015-06-04
- Granted: 2016-09-06
- Earliest claimed: 2013-06-11
- Assignees: GE Healthcare Bio-Sciences AB
- Applicants: GE Healthcare Bio-Sciences AB
- Inventors: Gunnar Glad; Therese Graner; Nils Norrman
- Web page: https://patentgrep.com/patent/US9433923B2

## CPC

- B01J 20/291: Gel sorbents
- B01D 15/3804: {Affinity chromatography}
- B01D 15/3828: {Ligand exchange chromatography, e.g. complexation, chelation or metal interaction chromatography}
- B01D 15/3885: {using electrical or magnetic means}
- B01J 20/265: {modified or post-treated polymers (polymer carriers or substrates subjected to further impregnating or coating B01J20/3208)}
- B01J 20/28009: {Magnetic properties}
- B01J 20/288: Polar phases
- B01J 20/3212: {consisting of a polymer obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds}
- B01J 20/3251: {comprising at least two different types of heteroatoms selected from nitrogen, oxygen or sulphur}
- B01J 20/3274: {Proteins, nucleic acids, polysaccharides, antibodies or antigens}
- C07K 1/22: Affinity chromatography or related techniques based upon selective absorption processes
- C12N 9/1088: {Glutathione transferase (2.5.1.18)}
- B01J 2220/80: Aspects related to sorbents specially adapted for preparative, analytical or investigative chromatography
- C07K 2319/23: containing a GST-tag
- B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention relates to a novel non-cross linked chromatography media provided with glutathione ligands which may or may not be provided with magnetic particles. The chromatography media is used for production of an affinity media provided with gluthatione ligands for adsorption of GST-tagged proteins.

## Claims

### Claim 1

A method for isolating proteins from a sample comprising:
a. combining a sample comprising glutathione-S-transferase (GST) tagged proteins with an affinity chromatography media comprising non-crosslinked agarose coated magnetic beads having glutathione immobilized thereon; and
b. magnetically separating the GST tagged proteins bound to the affinity media from the sample, thereby isolating the proteins from the sample.

### Claim 2 (depends on 1)

The method of claim 1, wherein the GST tagged proteins are combined directly after cell culture to the chromatography media.

### Claim 3 (depends on 2)

The method of claim 2, wherein the GST tagged proteins are concentrated after cell culture before combining with the chromatography media.

### Claim 4 (depends on 1)

The method of claim 1, wherein the affinity chromatography media provides improved isolation of GST tagged proteins over an affinity chromatography media produced from crosslinked agarose coated magnetic beads having glutathione immobilized thereon.