Process for producing porous cellulose beads using alkali aqueous solution, carrier for ligand immobilization, and adsorbent

Patent metadata

Field	Value
Publication number	US-2016244483-A1
Application number	US-201415025152-A
Country	US
Kind code	A1
Filing date	Sep 26, 2014
Priority date	Sep 27, 2013
Publication date	Aug 25, 2016
Grant date	—

Abstract

A process for producing porous cellulose beads of the present invention is characterized by comprising the steps of
a) mixing an alkali aqueous solution and cellulose to prepare cellulose micro dispersion at low temperature, b) adding water to the cellulose micro dispersion to prepare cellulose slurry, and d) bringing the cellulose slurry into contact with coagulation solvent.

A carrier for ligand immobilization of the present invention is characterized by being 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− V 2 /V 1 )×100

(wherein, V 1 indicates the gel volume of polysaccharide porous beads before shrinkage, and V 2 indicates the gel volume of polysaccharide porous beads after shrinkage).

First claim

1 : A process for producing porous cellulose beads, the process comprising:
mixing an alkali aqueous solution and cellulose at a temperature of from −5 to 10° C. such that a cellulose micro dispersion is prepared, adding water to the cellulose micro dispersion such that a cellulose slurry is prepared, and contacting the cellulose slurry with a coagulation solvent.

2 : The process according to claim 1 , further comprising
raising a temperature of the cellulose slurry, before contacting the cellulose slurry with the coagulation solvent.

3 : The process according to claim 1 , wherein an alkali concentration of the cellulose micro dispersion is from 8 wt % to 10 wt %.

4 : The process according to claim 1 , wherein an alkali concentration of the cellulose slurry is not less than 5 wt %.

5 : The process according to claim 1 , wherein a temperature of the cellulose slurry prepared in the adding is from 4° C. to 20° C.

6 : The process according to claim 1 , further comprising:
dispersing the cellulose slurry in a dispersion medium comprising a water-insoluble liquid such that a liquid-liquid dispersion containing droplets is formed, wherein the contacting of the cellulose slurry with the coagulation solvent comprises contacting the liquid-liquid dispersion with the coagulation solvent.

7 : The process according to claim 1 , wherein a concentration of the cellulose in the cellulose slurry is from 1 to 7 wt %.

8 : The process according to claim 1 , wherein the cellulose is at least one selected from the group consisting of regenerated cellulose, crystalline cellulose, microcrystalline cellulose, and cellulose acetate.

9 : The process according to claim 8 , wherein a degree of polymerization of the cellulose is not more than 1000.

10 : The process according to claim 6 , wherein the water-insoluble liquid is dichlorobenzene, hexane, ethyl acetate, a straight-chain saturated fatty acid having 6 to 12 carbons, an unsaturated fatty acid having 16 to 24 carbons, an animal fat or a vegetable oil having a melting point of not more than 100° C., a hydrogenated animal fat or a vegetable oil, a fractionated oil prepared by fractionating and purifying a high-melting point fraction of an animal fat or a vegetable oil or a hydrogenated animal fat or a vegetable oil, an unsaturated fatty acid triglyceride, an edible wax, a fat or an oil from microalgae, a fat or an oil from microorganisms, a medium-chain fatty acid triglyceride, or an unsaturated fatty acid triglyceride.

11 : The process according to claim 1 , wherein the coagulation solvent comprises an alcohol or a glycol.

12 : The process according to claim 11 , wherein the alcohol is at least one selected from the group consisting of isobutanol, 2-butanol, sec-butanol, 2-methyl-2-propanol, 1-propanol, 2-propanol, ethanol, and methanol.

13 : The process according to claim 11 , wherein the glycol is at least one selected from the group consisting of glycerol, ethylene glycol, and propylene glycol.

14 : Porous cellulose beads produced by the process according to claim 1 , wherein the porous cellulose beads have an exclusion limit molecular weight of from 1.0×10 6 to 1.0×10 11 .

15 : The porous cellulose beads according to claim 14 , wherein the porous cellulose beads have a median particle diameter of from 50 μm to 100 μm.

16 : 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.

17 : The carrier according to claim 16 , wherein the shrinking comprises contacting the polysaccharide porous beads with a water-soluble organic solvent and alkali water.

18 : The carrier according to claim 17 , 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.

19 : The carrier according to claim 18 , wherein process further comprises additionally contacting the beads obtained by the crosslinking with a crosslinking agent and alkali water at least one time.

20 : The carrier according to claim 16 , wherein the polysaccharides is cellulose or agarose.

21 : The carrier according to claim 17 , 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.

22 : The carrier according to claim 19 , wherein an alcohol solvent is not used in the additional crosslinking step.

23 : An adsorbent, obtained by immobilizing a ligand on the carrier according to claim 16 .

24 : The adsorbent according to claim 23 , wherein the ligand is an affinity ligand.

25 : The adsorbent according to claim 24 , wherein the affinity ligand is protein A, protein G, or protein L.

26 : A method for purifying an antibody by affinity chromatography, the method comprising:
contacting a source material with the adsorbent according to claim 24 , such that an antibody is adsorbed; 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.

Assignees

• Kaneka Corp

Inventors

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

Classifications

• C08J2301/02

  Cellulose; Modified cellulose · CPC title

• B01J20/3212

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

• B01J20/3293

  Coatings on a core, the core being particle or fiber shaped, e.g. encapsulated particles, coated fibers · CPC title

• C08L1/12

  Cellulose acetate · CPC title

• C08L2205/18

  Spheres · CPC title