Packing material for affinity chromatography

• US 8,846,877 B2
• Filed: 09/24/2009
• Issued: 09/30/2014
• Est. Priority Date: 09/25/2008
• Status: Expired due to Fees

• Current Assignee: JSR Corporation
• Original Assignee: JSR Corporation
• Inventors: Kawai, Takahiro, Tamori, Kouji, Fukuta, Tetsuo, Miyaji, Masaaki, Wang, Yong, Abe, Takayoshi, Okano, Yuusuke, Momiyama, Masaki
• Primary Examiner(s): Haq, Shafiqul
• Application Number:
  US13/121,124
• Publication Number:
  US 20110262748A1
• Time in Patent Office: 1,832 Days
• US Class Current: 530/413
• 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;

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

the ligand is bound to the porous mother particle by (i) utilizing the epoxy group comprised in the porous mother particles as a ligand binding site, (ii) activating an alcoholic hydroxyl group produced by ring-opening of the epoxy group comprised in the porous mother particles, and binding the ligand to the porous mother particles, or (iii) forming a linker that extends from the epoxy group comprised in the porous mother particles or a group produced by ring-opening of the epoxy group, and binding the ligand to the porous mother particles via the linker; and

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, 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.

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 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.

4. The affinity chromatography packing material of claim 1, 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, provided that a terminal of the site comprising 4 to 20 consecutive histidine residues binds to r, and

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

5. The affinity chromatography packing material of claim 1, 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.

6. 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.

7. 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.

8. 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.

9. 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.

10. 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.

11. 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.

12. 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.

13. The affinity chromatography packing material of claim 10, 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.

14. 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.

15. The affinity chromatography packing material of claim 10, 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.