PACKING MATERIAL FOR AFFINITY CHROMATOGRAPHY

• US 20110262748A1
• Filed: 09/24/2009
• Published: 10/27/2011
• Est. Priority Date: 09/25/2008
• Status: Active Grant

• Current Assignee: JSR Corporation
• Original Assignee: JSR Corporation
• Inventors: Wang, Yong, Tamori, Kouji, Momiyama, Masaki, Miyaji, Masaaki, Kawai, Takahiro, Okano, Yuusuke, Fukuta, Tetsuo, Abe, Takayoshi
• Granted Patent: US 8,846,877 B2
• Time in Patent Office: Days
• US Class Current: 428/357
• CPC Class Codes: B01J 20/286 Phases chemically bonded to a substrate, e.g. to silica or to polymers
• CPC Class Codes: B01J 20/321 consisting of a polymer obtained by reactions involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3219 involving a particular spacer or linking group, e.g. for attaching an active group
• CPC Class Codes: B01J 20/3274 Proteins, nucleic acids, polysaccharides, antibodies or antigens
• CPC Class Codes: C07K 1/22 Affinity chromatography or related techniques based upon selective absorption processes
• CPC Class Codes: Y10T 428/29 Coated or structually defined flake, particle, cell, strand, strand portion, rod, filament, macroscopic fiber or mass thereof

Abstract

An affinity chromatography packing material includes porous mother particles that include a copolymer of a monomer mixture including a crosslinkable vinyl monomer and an epoxy group-containing vinyl monomer, a ligand being bound to the porous mother particles, and the porous mother particles including a ring-opening epoxy group produced by ring-opening of the epoxy group included in the porous mother particles.

Claims

1. An affinity chromatography packing material, comprising:

at least one porous mother particle that comprises a copolymer of a monomer mixture comprising a crosslinkable vinyl monomer and an epoxy group-comprising vinyl monomer,

wherein a ligand is bound to the porous mother particle, and

the porous mother particle comprises a ring-opening epoxy group produced by ring-opening of the epoxy group comprised in the porous mother particles.

2. The affinity chromatography packing material of claim 1, wherein the ring-opening epoxy group is a substituted or unsubstituted 2,3-dihydroxypropyl group.

3. The affinity chromatography packing material of claim 1, wherein the ligand is a protein that comprises an immunoglobulin-binding domain of protein A.

4. The affinity chromatography packing material of claim 3, wherein the immunoglobulin-binding domain of protein A is at least one immunoglobulin-binding domain selected from the group consisting of an A-domain, a B-domain, a C-domain, a D-domain, an E-domain, and a Z-domain.

5. The affinity chromatography packing material of claim 1, wherein the ligand is an immunoglobulin-binding protein of formula (1),

R—

R² 

 

(1)

wherein

R represents an amino acid sequence that comprises 4 to 300 amino acids and a site including 4 to 20 consecutive histidine residues (SEQ ID NO;

     19), and

R² represents an amino acid sequence that comprises 50 to 500 amino acids and at least one immunoglobulin-binding domain of protein A, provided that a terminal of the immunoglobulin-binding domain binds to R.

6. The affinity chromatography packing material of claim 5, wherein R—

in formula (1) is a group of formula (2),

R¹-r- 

 

(2)

wherein

R¹ represents an amino acid sequence that comprises 4 to 100 amino acids and a site comprising 4 to 20 consecutive histidine residues (SEQ ID NO;

     19), provided that a terminal of the site comprising 4 to 20 consecutive histidine residues (SEQ ID NO;

     19) binds to r, and

r represents an arbitrary amino acid sequence that comprises 7 to 200 amino acids and a TEV domain.

7. The affinity chromatography packing material of claim 5, wherein at least one selected from the group consisting of the amino acid sequence represented by R and the amino acid sequence represented by R² in formula (1), comprises a domain t that comprises 1 to 50 amino acids and one amino acid selected from the group consisting of lysine, arginine, and cysteine.

8. The affinity chromatography packing material of claim 1, wherein the ligand is an immunoglobulin-binding protein of formula (3),

R²—

R 

 

(3)

wherein

R represents an amino acid sequence that comprises 4 to 300 amino acids and a site comprising 4 to 20 consecutive histidine residues (SEQ ID NO;

     19), and

R² represents an amino acid sequence that comprises 50 to 500 amino acids and at least one immunoglobulin-binding domain of protein A, provided that a terminal of the immunoglobulin-binding domain binds to R.

9. The affinity chromatography packing material of claim 8, wherein —

R in formula (3) is a group of formula (4),

-r-R¹ 

 

(4)

wherein

R¹ represents an amino acid sequence that comprises 4 to 100 amino acids and a site comprising 4 to 20 consecutive histidine residues (SEQ ID NO;

     19), provided that a terminal of the site comprising 4 to 20 consecutive histidine residues (SEQ ID NO;

     19) binds to r, and

r represents an arbitrary amino acid sequence that comprises 7 to 200 amino acids and a TEV domain.

10. The affinity chromatography packing material of claim 8, wherein at least one selected from the group consisting of the amino acid sequence represented by R and the amino acid sequence represented by R² in formula (3), comprises a domain t that comprises 1 to 50 amino acids and one amino acid selected from the group consisting of lysine, arginine, and cysteine.

11. The affinity chromatography packing material of claim 1, wherein the monomer mixture comprises:

20 to 50 wt % of the crosslinkable vinyl monomer; and

50 to 80 wt % of the epoxy group-comprising vinyl monomer.

12. The affinity chromatography packing material of claim 1, wherein the at least one porous mother particle has a volume average particle size of 20 to 80 micrometers.

13. The affinity chromatography packing material of claim 1, wherein the at least one porous mother particle has a volume average particle size of 30 to 60 micrometers.

14. The affinity chromatography packing material of claim 1, wherein the at least one porous mother particle has a specific surface area of 50 to 150 m²/g.

15. The affinity chromatography packing material of claim 1, wherein the at least one porous mother particle has a specific surface area of 80 to 120 m²/g.

16. The affinity chromatography packing material of claim 1, wherein the at least one porous mother particle has a volume average pore size of 100 to 400 nm.

17. The affinity chromatography packing material of claim 1, wherein the ring-opening epoxy group comprises a substituted 2,3-dihydroxypropyl group, wherein the substituted 2,3-dihydroxypropyl group is produced by ring-opening of the epoxy group comprised in the at least one porous mother particle with a mercapto group-comprising blocking agent.

18. The affinity chromatography packing material of claim 15, wherein the substituted 2,3-dihydroxypropyl group is produced by ring-opening of the epoxy group comprised in the at least one porous mother particle with mercaptoethanol.

19. The affinity chromatography packing material of claim 1, wherein the ring-opening epoxy group comprises a substituted 2,3-dihydroxypropyl group, wherein the substituted 2,3-dihydroxypropyl group is produced by ring-opening of the epoxy group comprised in the at least one porous mother particle with an amino group-comprising blocking agent.

20. The affinity chromatography packing material of claim 15, wherein the substituted 2,3-dihydroxypropyl group is produced by ring-opening of the epoxy group comprised in the at least one porous mother particle with monoethanolamine.