Carrier for ligand immobilization

Patent metadata

Field	Value
Publication number	US-10457705-B2
Application number	US-201816227688-A
Country	US
Kind code	B2
Filing date	Dec 20, 2018
Priority date	Sep 27, 2013
Publication date	Oct 29, 2019
Grant date	Oct 29, 2019

Abstract

A carrier for ligand immobilization obtained by shrinking polysaccharide porous beads not less than 10% by a shrinkage rate defined by the following formula, and crosslinking the polysaccharide porous beads: Shrinkage rate (%)=(1−V2/V1)×100 (wherein, V1 indicates the gel volume of polysaccharide porous beads before shrinkage, and V2 indicates the gel volume of polysaccharide porous beads after shrinkage).

First claim

The invention claimed is:

1. A carrier for ligand immobilization, obtained by a process comprising shrinking polysaccharide porous beads such that a shrinkage rate is not less than 10%, and crosslinking the polysaccharide porous beads,
wherein
Shrinkage rate (%)=(1− V 2 /V 1 )×100

wherein V 1 represents a gel volume of the polysaccharide porous beads before the shrinking, and V 2 represents a gel volume of the polysaccharide porous beads after the shrinking.

2. The carrier according to claim 1 , wherein the shrinking comprises contacting the polysaccharide porous beads into contact with a water-soluble organic solvent and alkali water.

3. The carrier according to claim 2 , wherein the shrinking is carried out in the presence of a crosslinking agent such that the polysaccharide porous beads are crosslinked during the shrinking, or the crosslinking is carried out after the shrinking such that shrunk polysaccharide porous beads obtained by the shrinking are crosslinked.

4. The carrier according to claim 3 , wherein the process further comprises, after the crosslinking, additionally contacting the polysaccharide porous beads with a crosslinking agent and alkali water at least one time.

5. The carrier according to claim 1 , wherein the polysaccharide porous beads comprise a polysaccharide which is cellulose or agarose.

6. The carrier according to claim 2 , wherein the water-soluble organic solvent is at least one selected from the group consisting of an alcohol solvent, a sulfoxide solvent, an amide solvent, a ketone solvent, and an ether solvent.

7. The carrier according to claim 4 , wherein an alcohol solvent is not used in the additional contacting crosslinking.

8. An adsorbent, obtained by immobilizing a ligand on the carrier according to claim 1 .

9. The adsorbent according to claim 8 , wherein the ligand is an affinity ligand.

10. The adsorbent according to claim 9 , wherein the affinity ligand is protein A, protein G, or protein L.

11. A method for purifying an antibody by affinity chromatography, the method comprising:
contacting a source material with the adsorbent according to claim 9 , such that an antibody is adsorbed on the adsorbent;
washing the antibody adsorbed on the adsorbent;
adding an eluent such that the antibody is liberated from the adsorbent; and
collecting the liberated antibody from the eluent.

12. The carrier according to claim 1 , wherein the polysaccharide porous beads have an exclusion limit molecular weight of 1.0×10 5 to 1.0×10 12 .

13. The carrier according to claim 1 , wherein the polysaccharide porous beads have an exclusion limit molecular weight of 1.0×10 6 to 1.0×10 11 .

14. The carrier according to claim 2 , wherein the water-soluble organic solvent is at least one selected from the group consisting of methanol, ethanol, propanol, dimethyl sulfoxide, dimethyl formamide, dimethyl acetamide, N-methylpyrrolidone, acetone, dioxane, and tetrahydrofuran.

15. The carrier according to claim 1 , wherein the process comprises shrinking the polysaccharide porous beads such that the shrinkage rate is 10% to 60%.

16. The carrier according to claim 1 , wherein the process comprises shrinking the polysaccharide porous beads such that the shrinkage rate is 20% to 50%.

17. The carrier according to claim 1 , wherein the carrier has a 20% compressive stress of 0.06 MPa to 0.28 MPa.

18. The carrier according to claim 1 , wherein the carrier has a 20% compressive stress of 0.072 MPa to 0.20 MPa.

19. The carrier according to claim 1 , wherein the carrier has a 20% compressive stress of 0.092 MPa to 0.20 MPa.

20. The carrier according to claim 1 , wherein the carrier has a particle diameter of 10 μm to 200 μm.

Assignees

• Kaneka Corp

Inventors

• Okubo Takahiro
• Kawai Yoshikazu
• Hirano Masaru
• Konoike Fuminori
• Karasugi Keiichi
• Honda Tatsuya

Classifications

• C08B15/08

  Fractionation of cellulose, e.g. separation of cellulose crystallites · CPC title

• B01J20/28016

  Particle form · CPC title

• B01J20/3212

  consisting of a polymer obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds · CPC title

• B01J20/286

  Phases chemically bonded to a substrate, e.g. to silica or to polymers · CPC title

• C08B15/005

  Crosslinking of cellulose derivatives · CPC title