Ligand-binding fiber and cell culture substrate using said fiber

Patent metadata

Field	Value
Publication number	US-10724028-B2
Application number	US-201515523157-A
Country	US
Kind code	B2
Filing date	Oct 30, 2015
Priority date	Oct 31, 2014
Publication date	Jul 28, 2020
Grant date	Jul 28, 2020

Abstract

The invention provides a ligand-bonded fiber in which a ligand having affinity for a cell membrane receptor is immobilized on a fiber precursor, and a cell culture substrate capable of repeating ex vivo amplification of a cell expressing a cell membrane receptor by using the ligand-bonded fiber.

First claim

The invention claimed is:

1. A ligand-bonded fiber comprising a ligand having affinity for a cell membrane receptor, and a fiber precursor bonded to the ligand,
wherein
the cell membrane receptor is a thrombopoietin (TPO) receptor and
the fiber precursor comprises a polymer compound comprising
(a) a unit structure represented by formula (1):

wherein
R 1 is a hydrogen atom or a methyl group,
Q 1 is an ester bond or amide bond,
R 2 is an alkyl group having 1-10 carbon atoms or an aromatic hydrocarbon group having 6-10 carbon atoms, wherein at least one hydrogen atom is substituted by a hydroxy group, and
(b) a unit structure represented by formula (2):

wherein
R 3 is a hydrogen atom or a methyl group, and
Q 2 is an active ester group.

2. The ligand-bonded fiber according to claim 1 , wherein the ligand has an amino group, and the amino group is bonded to the fiber precursor through the active ester group Q 2 .

3. The ligand-bonded fiber according to claim 1 , wherein Q 2 is represented by formula (3):

wherein Q 3 is an N-succinimide group, a p-nitrophenyl group or a pentafluorophenyl group.

4. The ligand-bonded fiber according to claim 1 , wherein the fiber precursor further comprises a crosslinking agent and an acid compound.

5. The ligand-bonded fiber according to claim 1 , wherein the fiber precursor is produced by spinning a composition for producing a fiber precursor, which composition comprising the polymer compound, a crosslinking agent, an acid compound and a solvent.

6. The ligand-bonded fiber according to claim 5 , wherein the fiber precursor is produced by spinning the composition for producing a fiber precursor, on a surface-treated substrate.

7. The ligand-bonded fiber according to claim 1 , wherein the polymer compound has a weight average molecular weight of 1,000-1,000,000.

8. The ligand-bonded fiber according to claim 1 , wherein the fiber precursor is produced by heating at 70-300° C.

9. The ligand-bonded fiber according to claim 1 , wherein the ligand is a compound represented by formula (4):

wherein
X 1 is a 3,4-dichlorophenyl group, a 4-trifluoromethylphenyl group or a 4-t-butylphenyl group,
X 2 is an optionally substituted amino group,
L 1 is a single bond or —CH 2 —C 6 H 4 —,
L 2 is a single bond or —CONH—, and
L 3 is an alkylene group having 2-6 carbon atoms.

10. The ligand-bonded fiber according to claim 9 , wherein X 1 is a 4-t-butylphenyl group, and X 2 is an amino group.

11. A cell culture substrate comprising the ligand-bonded fiber according to claim 1 .

12. The ligand-bonded fiber according to claim 4 , wherein the fiber precursor is produced by heating at 70-300° C.

13. The ligand-bonded fiber according to claim 4 , wherein the ligand is a compound represented by formula (4):

wherein
X 1 is a 3,4-dichlorophenyl group, a 4-trifluoromethylphenyl group or a 4-t-butylphenyl group,
X 2 is an optionally substituted amino group,
L 1 is a single bond or —CH 2 —C 6 H 4 —,
L 2 is a single bond or —CONH—, and
L 3 is an alkylene group having 2-6 carbon atoms.

14. The ligand-bonded fiber according to claim 13 , wherein X 1 is a 4-t-butylphenyl group, and X 2 is an amino group.

15. A cell culture substrate comprising the ligand-bonded fiber according to claim 4 .

Assignees

• Nissan Chemical Ind Ltd

Inventors

• Umezaki Makiko
• Kishioka Takahiro
• Nishino Taito
• Aihara Ayako
• Iwamoto Shunsuke
• Sakuma Daisuke

Classifications

• C12N11/087 Primary

  Acrylic polymers · CPC title

• C12N11/082

  obtained by reactions only involving carbon-to-carbon unsaturated bonds · CPC title

• C12N11/089

  obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds · CPC title

• C12N1/00

  Microorganisms; Compositions thereof (medicinal preparations containing material from protozoa, bacteria or viruses A61K35/66, from algae A61K36/02, from fungi A61K36/06; preparing medicinal bacterial antigen or antibody compositions, e.g. bacterial vaccines, A61K39/00); Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor · CPC title

• C12M1/00

  Apparatus for enzymology or microbiology · CPC title