Methods of concentrating ligands and active membranes used therefor

• US 4,693,985 A
• Filed: 08/21/1984
• Issued: 09/15/1987
• Est. Priority Date: 08/21/1984
• Status: Expired due to Term

• Current Assignee: Pall Corporation (Danaher Corporation)
• Original Assignee: Pall Corporation (Danaher Corporation)
• Inventors: Martin, Jerold, Shirley, Brenda, Schriefer, Judy, Degen, Peter J.
• Primary Examiner(s): NOT, DEFINED
• Application Number:
  US06/642,899
• Time in Patent Office: 1,120 Days
• Field of Search: 436/531, 436/532, 436/548, 436/824, 436/827, 436/828, 435/180, 435/182, 435/181, 525/420.5, 210/198.2, 210/493.1, 210/500.38, 210/502.1, 210/635
• US Class Current: 436/531
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: B01D 67/0093 Chemical modification
• CPC Class Codes: B01D 69/141 Heterogeneous membranes, e.g. containing dispersed material; Mixed matrix membranes
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/545 Synthetic resin
• CPC Class Codes: Y10S 436/824 Immunological separation techniques
• CPC Class Codes: Y10S 436/827 Lectins
• CPC Class Codes: Y10S 436/828 Protein A

Abstract

A chemically active membrane having a large surface area is provided in which a hydrophilic, microporous, skinless, polyamide membrane is chemically bound to a residue of an activating agent which is capable of reacting with a biologically active material.

The chemically active membrane, formed by reacting a hydrophilic, microporous, skinless, polyamide membrane with an activating agent may be used to prepare a biologically active membrane having a large surface area which comprises an acceptor molecule such as a monoclonal antibody, a polyclonal antibody, an antigenic substance, a glycoprotein, Protein A, a lectin, a carbohydrate, an enzyme substrate, a cofactor, an inhibitor, a hormone, an IgG class of immunoglobulin, a carrier protein, a receptor, heparin, a coagulation factor, or a histone covalently bound to the hydrophilic, microporous, skinless, polyamide membrane by reacting the chemically activate membrane with the acceptor molecule.

Claims

1. A method of preparing a biologically active membrane having a large surface area comprising the steps of:

reacting a hydrophilic, microporous, skinless polyamide membrane with an activating agent to form a chemically activated membrane, and

reacting said chemically activated membrane with an acceptor molecule to form a biologically active membrane, said acceptor molecule being a member selected from the group consisting of a monoclonal antibody, a polyclonal antibody, an antigenic substance, a glycoprotein, Protein A, a lectin, a carbohydrate, an enzyme substrate, a cofactor, an inhibitor, a hormone, an IgG class of immunoglobulin, a carrier protein, a receptor, heparin, a coagulation factor, and a histone.

2. The method of claim 1 wherein said polyamide membrane comprises polyhexamethylene adipamide.

3. The method of claim 2 wherein said membrane is formed from a material modified to include controlled surface properties.

4. The method of claim 2 wherein said modified polyamide membrane includes a high concentration of carboxyl moieties.

5. The method of claim 2 wherein said activating agent includes at least one functional group capable of reacting with moieties on the surface of said polyamide membrane to form an activated membrane having a functional group capable of reacting with said acceptor molecule.

6. The method of claim 2 wherein said acceptor molecule is a monoclonal antibody.

7. The method of claim 2 wherein said activating agent comprises trichloro-s-triazine.

8. The method of claim 2 wherein said activating agent comprises N,N'"'"'-dicyclohexylcarbodiimide.

9. The method of claim 2 wherein a substrate is embedded in said membrane.

10. A method for preparing a biologically active membrane having a large surface area comprising the steps of:

reacting a hydrophilic, microporous, skinless, polyamide membrane with an activating agent to form a chemically activated membrane wherein said activating agent is either trichloro-s-triazine or N, N'"'"'-dicyclohexylcarbodiimide, and

reacting said chemically activated membrane with an acceptor molecule to form a biologically active membrane.

11. The method of claim 10 wherein said acceptor molecule is capable of reacting with a ligand to form a stable biospecific complex.

12. The method of claim 10 wherein said acceptor molecule is a proteinaceous substance.

13. The method of claim 11 wherein said acceptor molecule is a member selected from the group consisting of a monoclonal antibody, a polyclonal antibody, an antigenic substance, a glycoprotein, Protein A, a lectin, a carbohydrate, an enzyme substrate, a cofactor, an inhibitor, a hormone, an IgG class of immunoglobulin, a carrier protein, a receptor, heparin, a coagulation factor, and a histone.

14. A biologically active membrane having a large surface area comprising an acceptor molecule selected from the group consisting of a monoclonal antibody, a polyclonal antibody, an antigenic substance, a glycoprotein, Protein A, a lectin, a carbohydrate, an enzyme substrate, a cofactor, an inhibitor, a hormone, an IgG class of immunoglobulin, a carrier protein, a receptor, heparin, a coagulation factor, and a histone covalently bound to a hydrophilic, microporous, skinless polyamide membrane.

15. The biologically active membrane of claim 14 wherein said polyamide membrane comprises polyhexamethylene adipamide.

16. The biologically active membrane of claim 15 wherein said polyamide membrane has a methylene CH₂ :

amide NHCO ratio of from about 5;

1 to about 7;

1.

17. The biologically active membrane of claim 15 wherein said polyamide membrane is modified to include controlled surface properties.

18. The biologically active membrane of claim 17 wherein said controlled surface properties include a high concentration of carboxyl moieties.

19. The biologically active membrane of claim 15 wherein a residue of an activating agent covalently links said acceptor molecule to said hydrophilic, microporous, skinless polyamide membrane.

20. The biologically active membrane of claim 19 wherein said residue of an activating agent comprises a residue of trichloro-s-triazine.

26. The biologically active membrane of claim 15 wherein a substrate is embedded in said membrane.

27. A filter element comprising the biologically active membrane of claim 26 wherein said membrane is positioned intermediate porous support sheets forming thereby a membrane element and said membrane element is arranged in a tubular corrugated configuration having axially extending pleats, said tubular configured membrane element being placed intermediate a pair of concentric, cylindrical, rigid, perforated tubes, the ends of said tubular configured membrane element and said concentric cylindrical tubes being closed off with end caps forming a fluid-tight seal between said tubular configured membrane element and said concentric perforated tubes.

21. A biologically active membrane having a large surface area comprising an acceptor molecule covalently bound to a hydrophilic, microporous, skinless, polyamide membrane through a residue of trichloro-s-triazine.

22. The biologically active membrane of claim 21 wherein said acceptor molecule is capable of reacting with a ligand to form a stable biospecific complex.

23. The biologically active membrane of claim 21 wherein said biologically active material is a proteinaceous substance.

24. The biologically active membrane of claim 21 wherein said acceptor molecule is a member selected from the group consisting of a monoclonal antibody, a polyclonal antibody, an antigenic substance, a glycoprotein, Protein A, a lectin, a carbohydrate, an enzyme substrate, a cofactor, an inhibitor, a hormone, an IgG class of immunoglobulin, a carrier protein, a receptor, heparin, and a coagulation factor.

25. The biologically active membrane of claim 21 wherein said acceptor molecule is a monoclonal antibody.

28. A method for recovering a ligand from a solution including passing a ligand solution through a biologically active membrane comprising an acceptor molecule selected from the group consisting of a monoclonal antibody, a polyclonal anitbody, an antigenic substance, a glycoprotein, Protein A, a lectin, a carbohydrate, an enzyme substrate, a cofactor, an inhibitor, a hormone, an IgG class of immunoglobulins, a carrier protein, a receptor, heparin, a coagulation factor, and a histone covalently bound to a hydrophilic, microporous, skinless polyamide membrane and binding said ligand to said biologically active membrane.

29. The method of claim 28 wherein said polyamide membrane comprises polyhexamethylene adipamide.

30. The method of claim 29 wherein said polyamide membrane is formed from a material modified to include controlled surface properties.

31. The method of claim 30 wherein said polyamide membrane includes a high concentration of carboxyl moieties.

32. The method of claim 29 wherein a substrate is embedded in said polyamide membrane.

33. The method of claim 29 wherein said ligand is a proteinaceous substance.

34. The method of claim 29 wherein said ligand is a member of a group consisting of an antigenic substance, a monoclonal antibody, a polyclonal antibody, a lectin, a glycoprotein, a carbohydrate, Protein A, an enzyme, a carrier protein, a receptor, a hormone, a coagulation factor, heparin, a nucleic acid, and a polynucleotide.

39. The method of claim 29 wherein said acceptor molecule is chemically bound to said polyamide membrane through a residue of an activating agent.

40. The method of claim 39 wherein said activating agent comprises trichloro-s-triazine.

41. The method of claim 29 wherein the bound ligand is eluted from said biologically active membrane.

42. The method of claim 29 wherein a substrate is embedded in said membrane.

43. The method of claim 42 wherein said membrane is positioned intermediate between porous support sheets, forming thereby a membrane element, and said membrane element is arranged in a tubularical corrugated configuration having axially extending pleats, said tubular configured membrane element being placed intermediate a pair of concentric, cylindrical, rigid, perforated tubes, the ends of said concentric, cylindrical tubes and said tubular configured membrane element being closed of with end caps to form a fluid-tight seal between said tubular configured membrane element and said concentric perforated tubes.

35. A method for recovering a ligand from a solution including passing a ligand solution through a biologically active membrane comprising an acceptor molecule covalently bound to a hydrophilic, microporous, skinless, polyamide membrane through a residue of trichloro-s-triazine and binding said ligand to said biologically active membrane.

36. The method of claim 35 wherein said acceptor molecule is a member selected from the group consisting of a monoclonal antibody, a polyclonal antibody, an antigenic substance, a glycoprotein, Protein A, a lectin, a carbohydrate, an enzyme substrate, a cofactor, an inhibitor, a hormone, an IgG class of immunoglobulins, a carrier protein, a receptor, heparin, a coagulation factor, and a histone.

37. The method of claim 35 wherein said acceptor molecule is a proteinaceous substance.

38. The method of claim 36 wherein said acceptor molecule is a monoclonal antibody.

44. A method for preparing a chemically activated membrane comprising reacting a hydrophilic, microporous, skinless polyamide membrane with an activating agent derived from trichloro-s-triazine or N, N'"'"'-dicyclohexylcarbodiimide to form a chemically activated membrane.

45. The method of claim 44 wherein said polyamide membrane comprises polyhexamethylene adipamide.

46. The method of claim 45 wherein said membrane is formed from a material modified to include controlled surface properties.

47. The method of claim 46 wherein said modified polyamide membrane includes a high concentration of carboxyl moieties.

48. The method of claim 45 wherein said polyamide membrane has a methylene CH₂ :

amide NHCO ratio of from about 5;

1 to about 7;

1.

49. The method of claim 45 wherein said polyamide membrane is modified to include controlled surface properties.

50. The method of claim 49 wherein said controlled surface properties include a high concentration of carboxyl moieties.

51. The method of claim 45 wherein a substrate is embedded in said membrane.

52. The method of claim 51 wherein said membrane is positioned intermediate porous support sheets, forming thereby a membrane element, and said membrane element is arranged in a tubular corrugated configuration having axially extending pleats, said tubular configured membrane element being placed intermediate a pair of concentric, cylindrical, rigid, perforated tubes, the ends of said tubular configured membrane element and said concentric, cylindrical tubes being closed off with end caps to form a fluid-tight seal between said tubular configured membrane element and said concentric perforated tubes.

53. A chemically active membrane having a large surface area comprising a hydrophilic, microporous, skinless, polyamide membrane chemically bound to a residue of an activating agent, which residue is derived from trichloro-s-triazine or N, N'"'"' dicyclohexylcarbodiimide and is capable of reacting with a biologically active material.

54. The chemically active membrane of claim 53 wherein said polyamide membrane comprises polyhexamethylene adipamide.

55. The chemically active membrane of claim 54 wherein said polyamide membrane has a methylene CH₂ :

amide NHCO ratio of about 5;

1 to about 7;

1.

56. The chemically active membrane of claim 54 wherein said polyamide membrane is modified to include controlled surface properties.

57. The chemically active membrane of claim 56 wherein said polyamide membrane includes a high concentration of carboxyl moieties at the surface of said membrane.

58. The chemically active membrane of claim 54 wherein said residue is derived from trichloro-s-triazine.

59. The chemically active membrane of claim 54 wherein said residue is derived from N,N'"'"'-dicyclohexylcarbodiimide.

60. The chemically active membrane of claim 54 wherein a substrate is embedded in said membrane.

61. A filter element comprising the chemically active membrane of claim 60 wherein said membrane is positioned intermediate porous support sheets, forming thereby a membrane element, and said membrane element is arranged in a tubular corrugated configuration having axially extending pleats, said tubular configured membrane element being placed intermediate a pair of concentric, cylindrical, rigid, perforated tubes, the open ends of said tubular configured membrane element and said concentric, cylindrical tubes being closed off with end caps forming a fluid-tight seal between said tubular configured membrane element and said concentric, cylindrical tubes.