# US 11,439,926 B2 · Chromatography medium

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- Number: US 11,439,926 B2
- Document ID: US11439926B2
- Status: Granted
- Kind: B2
- Application number: 16/701,942
- Filed: 2019-12-03
- Published: 2020-04-02
- Granted: 2022-09-13
- Earliest claimed: 2008-04-22
- Assignees: Cytiva BioProcess R&D AB
- Applicants: Cytiva BioProcess R&D AB
- Inventors: Jan Bergstrom; Gunnar Glad; Bo-Lennart Johansson; Jean-Luc Maloisel; Nils Norrman; Tobias E. Soderman
- Web page: https://patentgrep.com/patent/US11439926B2

## CPC

- B01D 15/327: {with hydrophobic interaction}
- B01J 20/28009: {Magnetic properties}
- B01J 20/285: based on polymers
- B01J 20/287: Non-polar phases; Reversed phases
- B01J 20/3212: {consisting of a polymer obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds}
- B01J 20/3293: {Coatings on a core, the core being particle or fiber shaped, e.g. encapsulated particles, coated fibers}
- B01J 39/26: Cation exchangers for chromatographic processes
- C07K 1/20: Partition-, reverse-phase or hydrophobic interaction chromatography
- C07K 16/00: Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- G01N 1/405: {by adsorption or absorption}
- C12N 2760/16151: Methods of production or purification of viral material
- 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)
- B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- 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)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

The present invention is within the field of chromatography. More precisely, it relates to a novel chromatography medium, namely a hydrophobic medium provided with different lids excluding molecules over a certain size due to the porosity of the hydrophobic medium and/or the porosity of the lid. The invention also relates to use of the separation medium for purification of large molecules, which do not enter the separation medium, as well as small molecules, which enter the separation medium and are eluted from there.

## Claims

### Claim 1

An agarose-based separation medium, comprising,
a hydrophobic porous core entity with octylamine ligands having a ligand density of greater than 90 μmole/mL; and
a porous hydrophilic lid covering the whole outer part of the core entity,
wherein the lid only allows molecules under 60,000D to penetrate and absorb onto the hydrophobic octyl ligands of said core entity of the separation medium,
wherein the hydrophobic core entity allows adsorption of molecules under 60,000D at low ionic strength 0-1M in the pH interval 2-11.

### Claim 2 (depends on 1)

The separation medium of claim 1, wherein the hydrophobic core entity allows adsorption of molecules under 1,000D.

### Claim 3 (depends on 1)

The separation medium of claim 1, wherein the hydrophilic lid comprises ligands that reversibly bind the molecules within a sample.

### Claim 4 (depends on 1)

The separation medium of claim 1, wherein magnetic particles are incorporated into the core entity.

### Claim 5 (depends on 1)

The separation medium of claim 1, wherein a dextran hydrogel is provided in the lid to fill the pores of the lid and thereby further decrease and adjust the pore size of the lid.

### Claim 6 (depends on 1)

The separation medium of claim 1, wherein at least one of the lid and core entity are functionalized by layer activation.