Protein adsorbent

• US 4,246,351 A
• Filed: 10/29/1979
• Issued: 01/20/1981
• Est. Priority Date: 10/28/1976
• Status: Expired due to Term

• Current Assignee: Asahi Kasei Kogyo Kabushiki Kaisha
• Original Assignee: Asahi Kasei Kogyo Kabushiki Kaisha
• Inventors: Ikeda, Akihiko, Takeda, Kunihiko, Miyake, Tetsuya, Mizuno, Masayuki
• Primary Examiner(s): Foelak, Morton
• Application Number:
  US06/088,927
• Time in Patent Office: 449 Days
• Field of Search: 435/180, 435/182, 435/815, 260/8, 260/112 R, 260/112 B, 260/122, 521/53
• US Class Current: 435/182
• CPC Class Codes: B01J 20/22 comprising organic material
• CPC Class Codes: C07K 1/16 by chromatography
• CPC Class Codes: C08J 9/286 the liquid phase being a solvent for the monomers but not for the resulting macromolecular composition, i.e. macroporous or macroreticular polymers
• CPC Class Codes: C12H 1/0424 with the aid of a polymer
• CPC Class Codes: C12N 11/082 obtained by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: C12N 11/087 Acrylic polymers
• CPC Class Codes: Y10S 210/905 Protein
• CPC Class Codes: Y10S 435/815 by sorption
• CPC Class Codes: Y10S 530/806 Antigenic peptides or proteins

Abstract

A novel protein adsorbent consisting essentially of a porous copolymer obtained by copolymerizing a monomer mixture comprising at least one cyano group-containing monomer and at least one cross-linkable monomer, said copolymer having an average pore diameter (d) of from 40 A to 9,000 A and a total pore volume of from 0.05√X ml to 1.5√X ml per gram of the copolymer in a dry state, in which X designates the weight proportion of said cross-linkable monomer expressed in terms of the percent by weight based on the total monomers. The present porous copolymer adsorbent has a remarkably high protein adsorbing capacity. The adorbent can advantageously be used for various purposes such as purification and separation of proteins, and removal of proteins. Copolymer-protein composites obtained by adsorption of proteins on the present porous copolymer also are industrially useful and have a wide variety of applications.

Claims

1. A method for adsorbing a protein from an aqueous solution or dispersion of the protein which comprises contacting an aqueous solution or a dispersion of a protein with a porous copolymer obtained by copolymerizing a monomer mixture comprising 2 to 98% by weight of at least one cyano group-containing monomer represented by the following general formula (A):

##STR6## wherein R stands for a hydrogen atom, an alkyl group having 1 to 10 carbon atoms, a halogen atom, a cyano group, a methoxy group, an acetoxyl group, or an unsubstituted or substituted phenyl group represented by the formula ##STR7## wherein R'"'"' stands for a hydrogen atom, an alkyl group having 1 to 10 carbon atoms, a halogen atom, a cyano group, a methoxy group or an acetoxyl group), and 2 to 98% by weight of at least one cross-linkable monomer having a plurality of CH₂ ═

C<

groups, said copolymer having an average pore diameter (d) of from 40 A to 9,000 A and a total pore volume of from 0.05 √

X ml to 1.5 √

X ml per gram of the copolymer in a dry state, in which X designates the weight proportion of said cross-linkable monomer expressed in terms of the percent by weight based on the total monomers, and being employed in an amount of 0.02 to 300 g per liter of said solution or said dispersion.

2. A method as set forth in claim 1 wherein the contacting of said solution or said dispersion with said porous copolymer is batchwise effected.

3. A method as set forth in claim 1 wherein the contacting of said solution or said dispersion with said porous copolymer is continuously effected.

4. A method as set forth in claim 1 wherein the contacting of said solution or said dispersion with said porous copolymer is effected at a temperature in the range of from 2°

C. to 95°

C.

5. A method as set forth in claim 2 wherein the contacting time is in the range of from 10 seconds to 500 hours.

6. A method as set forth in claim 3 wherein said solution or said dispersion is supplied at a space velocity in the range of from 0.01 hr^-1 to 1,000 hr^-1.

7. A method as set forth in claim 1 wherein said solution or said dispersion has a pH of from 1 to 11.

12. A method as set forth in claim 1 wherein the content of the cyano group-containing monomer is 4 to 95% by weight.

13. A method as set forth in claim 1 wherein the content of the cyano group-containing monomer is 6 to 90% by weight.

14. A method as set forth in claim 1 wherein the cyano group-containing monomer is a member selected from the group consisting of acrylonitrile, methacrylonitrile and mixtures thereof.

15. A method as set forth in claim 1 wherein the cross-linkable monomer is a monomer having 2 to 4 vinyl groups.

16. A method as set forth in claim 1 wherein the cross-linkable monomer is divinylbenzene.

17. A method as set forth in claim 1 wherein the cross-linkable monomer is of the formula ##STR10## wherein R₁ and R₂ each independently is hydrogen or methyl, and n is an integer from 1 to 30.

18. A method as set forth in claim 1 wherein the weight proportion of the cross-linkable monomer is 8 to 80% by weight.

19. A method as set forth in claim 1 wherein the average pore diameter (d) of said porous copolymer is in the range of from 50 A to 5,000 A.

20. A method as set forth in claim 1 wherein the volume of pores of said porous copolymer having a diameter of 0.5 d to 2 d is not more than 60% of the total pore volume.

21. A method as set forth in claim 1 wherein the total pore volume of said porous copolymer is in the range of from 0.2 √

X ml to 1.3 √

X ml per gram of the copolymer in a dry state.

22. A method as set forth in claim 1 wherein the protein has a molecular weight of from 1,000 to 1,000,000.

23. A method as set forth in claim 1 wherein the protein is albumin of human serum.

24. A method as set forth in claim 1 wherein the protein is α

-chymotrypsin.

25. A method as set forth in claim 1 wherein the protein is lipase.

26. A method as set forth in claim 1 wherein the protein is an enzyme.

27. A method as set forth in claim 1 wherein the protein is trypsin.

28. A method as set forth in claim 1 wherein the protein is chymotrypsin.

29. A method as set forth in claim 1 wherein the protein is an antigen.

30. A method as set forth in claim 1 wherein the protein is an antibody.

31. A method as set forth in claim 1 wherein said porous copolymer has a spherical shape.

32. A method as set forth in claim 31 wherein said porous copolymer is a product obtained by suspension polymerization which is carried out in an aqueous solution in the presence of a radical initiator.

33. A composite produced by the method of claim 1 which comprises the porous copolymer and the protein in an amount sufficient to exert activity of said protein adsorbed thereon.

34. A composite as set forth in claim 33 wherein the amount of said protein is at least 10 mg per gram of the copolymer in a dry state.

35. A composite as set forth in claim 34 wherein the amount of said protein is at least 20 mg per gram of the copolymer in a dry state.

36. A composite as set forth in claim 33 wherein the protein is an enzyme.

8. A composite comprising a porous copolymer and a protein in an amount sufficient to exert activity of said protein adsorbed thereon, said porous copolymer being a product obtained by copolymerizing a monomer mixture comprising 2 to 98% by weight of at least one cyano group-containing monomer represented by the following general formula (A):

##STR8## wherein R stands for a hydrogen atom, an alkyl group having 1 to 10 carbon atoms, a halogen atom, a cyano group, a methoxy group, an acetoxyl group, or an unsubstituted or substituted phenyl group represented by the formula ##STR9## (wherein R'"'"' stands for a hydrogen atom, an alkyl group having 1 to 10 carbon atoms, a halogen atom, a cyano group, a methoxy group or an acetoxyl group), and 2 to 98% by weight of at least one cross-linkable monomer having a plurality of CH₂ ═

C<

groups, said copolymer having an average pore diameter (d) of from 40 A to 9,000 A and a total pore volume of from 0.05 √

X ml to 1.5 √

X ml per gram of the copolymer in a dry state, in which X designates the weight proportion of said crosslinkable monomer expressed in terms of the percent by weight based on the total monomers.

9. A composite as set forth in claim 8 wherein the amount of the protein is at least 10 mg per gram of the copolymer in a dry state.

10. A composite as set forth in claim 9 wherein the amount of the protein is at least 20 mg per gram of the copolymer in a dry state.

11. A composite as set forth in claim 8, wherein the protein is an enzyme.