Materials and methods for the immobilization of bioactive species onto polymeric substrates

• US 5,897,955 A
• Filed: 08/21/1998
• Issued: 04/27/1999
• Est. Priority Date: 06/03/1996
• Status: Expired due to Term

• Current Assignee: W. L. Gore & Associates (W.L. Gore & Associates)
• Original Assignee: Gore Hybrid Technologies, Inc.
• Inventors: Drumheller, Paul D.
• Primary Examiner(s): Richter, Johann
• Assistant Examiner(s): MURRAY, JOSEPH H
• Application Number:
  US09/138,197
• Time in Patent Office: 249 Days
• Field of Search: 428/426, 428/422, 428/432, 435/7.92, 435/180, 435/181
• US Class Current: 428/422
• CPC Class Codes: A61L 27/34 Macromolecular materials
• CPC Class Codes: C07K 17/08 the carrier being a synthetic polymer
• CPC Class Codes: C12N 11/082 obtained by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: C12N 11/084 Polymers containing vinyl alcohol units
• CPC Class Codes: C12N 11/087 Acrylic polymers
• CPC Class Codes: C12N 11/096 Polyesters; Polyamides
• CPC Class Codes: C12N 2533/30 Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 2533/50 Proteins
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: Y10T 428/249958 Void-containing component is synthetic resin or natural rubbers
• CPC Class Codes: Y10T 428/31544 Addition polymer is perhalogenated

Abstract

The present invention is directed to support members having surfaces that are rendered hydrophilic for use as a substrate for the immobilization of bioactive species thereon. The hydrophilic surfaces are chemically stable on the support member. The surfaces are also chemically variable to provide a variety of chemically functional groups for immobilization of bioactive species thereto. The surfaces comprise polymeric surfactants attached onto the surfaces of a support member and covalently cross-linked thereon to form a first layer. Hydrophilic polymers are then attached to the first layer on the support member to form a second layer thereon. The second layer is used to enhance the hydrophilicity of a support member, as well as, to provide a substrate for immobilizing bioactive species thereto. Methods for forming the first and second layers are also provided. In addition, methods for attaching bioactive substances to the hydrophilic polymers are provided.

Claims

1. A material having bioactive species immobilized thereto, said material comprising:

a support member;

a first layer comprised of at least one species of a polymeric surfactant non-covalently attached to the support member, wherein the polymeric surfactant is a different chemical species than the support member, wherein the polymeric surfactant is cross-linked to itself in situ with a cross-linking agent; and

at least one type of bioactive species attached to the cross-linking agent.

2. The material of claim 1 wherein the support member comprises a non-porous polymer material.

3. The material of claim 2 wherein the non-porous polymer material is selected from a member of the group consisting of polytetrafluoroethylene, polyethylene, polypropylene, polyethyleneterephthalate, polystyrene, polysulfone, polycarbonate, and regenerated cellulose.

4. The material of claim 1 wherein the support member comprises a porous polymer material.

5. The material of claim 4 wherein the porous polymer material is selected from a member of the group consisting of porous polytetrafluoroethylene, porous polyethylene, porous polypropylene, porous polyethyleneterephthalate, porous polyurethane, porous silicone rubber, porous polystyrene, porous polysulfone, porous polyester, porous polyhydroxyacids, porous polycarbonate, porous polyimide, porous polyamide, porous polyamino acids, and porous regenerated cellulose.

7. The material of claim 1 wherein a spacer compound is interposed between the first layer and the bioactive species.

8. The material of claim 7 wherein the spacer compound is selected from a member of a group consisting of succinic acid, diaminohexane, glyoxylic acid, short chain polyethylene glycol, and glycine.

9. The material of claim 7 wherein the spacer compound is cleavable.

10. The material of claim 9 wherein the cleavable spacer compound is selected from the group consisting of polyhydroxyacids, polyanhydrides, polyamino acids, tartarates, and cysteine-linkers.

6. The material of claim I wherein the polymeric surfactant comprises a multifunctional copolymer comprised of at least one domain having a physicochemical affinity for the support member and at least one domain that is chemically reactive with the cross-linking agent.

11. A material having immobilized bioactive species comprising:

a hydrophobic support member;

a first layer comprised of at least one species of a polymeric surfactant non-covalently attached to the support member and cross-linked to itself in situ with a cross-linking agent; and

at least one type of bioactive species attached to the cross-linking agent.

12. The material of claim 11 wherein the polymeric surfactant comprises a multifunctional copolymer comprised of at least one domain having a physicochemical affinity for the support member and at least one domain that is chemically reactive with the cross-linking agent.

13. The material of claim 11 wherein a spacer compound is interposed between the first layer and the bioactive species.

14. The material of claim 13 wherein the spacer compound is selected from a member of a group consisting of succinic acid, diaminohexane, glyoxylic acid, short chain polyethylene glycol, and glycine.

15. The material of claim 13 wherein the spacer compound is cleavable.

16. The material of claim 15 wherein the cleavable spacer compound is selected from the group consisting of polyhydroxyacids, polyanhydrides, polyamino acids, tartarates, and cysteine-linkers.

32. The material of claim 14 wherein the spacer compound is cleavable.

33. The material of claim 16 wherein the cleavable spacer compound is selected from the group consisting of polyhydroxyacids, polyanhydrides, polyamino acids, tartarates, and cysteine-linkers.

17. A material for immobilization of bioactive species thereto, said material comprising:

a support member;

a first layer comprising at least one species of a polymeric surfactant non-covalently attached to the support member, wherein the polymeric surfactant is a different chemical species than the support member, and wherein the polymeric surfactant is cross-linked to itself in situ; and

a second layer comprising at least one species of a hydrophilic polymer covalently attached to the first layer, wherein the second layer has chemically reactive moieties for attaching bioactive species thereto.

18. The material of claim 17 wherein the support member comprises a non-porous polymer material.

19. The material of claim 18 wherein the non-porous polymer material is selected from a member of the group consisting of polytetrafluoroethylene, polyethylene, polypropylene, polyethyleneterephthalate, polystyrene, polysulfone, polycarbonate, polyethylene-co-vinyl acetate, polyamide, polyurethane, and regenerated cellulose.

20. The material of claim 17 wherein the support member is selected from a member of the group consisting of silk, wool, and leather.

21. The material of claim 17 wherein the support member comprises a porous polymer material.

22. The material of claim 21 wherein the porous polymer material is selected from a member of the group consisting of porous polytetrafluoroethylene, porous polyethylene, porous polypropylene, porous polyethyleneterephthalate, porous polyurethane, porous silicone rubber, porous polystyrene, porous polysulfone, porous polyester, porous polyhydroxyacids, porous polycarbonate, porous polyimide, porous polyamide, porous polyamino acids, and porous regenerated cellulose.

23. The material of claim 17 wherein the polymeric surfactant comprises a multifunctional copolymer comprised of at least one domain having a physicochemical affinity for the support member and at least one domain that is chemically reactive with a cross-linking agent.

24. The material of claim 17 wherein the hydrophilic polymer comprises chemically functional groups to which the first layer is covalently attached and additional chemically functional groups to which bioactive species can be attached.

25. The material of claim 17 wherein the polymeric surfactant of the first layer is selected from the group consisting of polyvinyl alcohol, polyethylene glycol, polypropylene glycol, dextran, agarose, alginate, polyacrylamide, polyglycidol, poly(vinyl alcohol-co-ethylene), poly(ethyleneglycolco-propyleneglycol), poly(vinyl acetate-co-vinyl alcohol), poly(tetrafluoroethylene-co-vinyl alcohol), poly(acrylonitrile-co-acrylamide), poly(acrylonitrile-co-acrylic acid-co-acrylamidine), polyacrylic acid, poly-lysine, polyethyleneimine, polyvinyl pyrrolidone, polyhydroxyethylmethacrylate, and polysulfone, and their copolymers, either alone or in combination.

26. The material of claim 17 wherein the hydrophilic polymer of the second layer is selected from the group consisting of polyvinyl alcohol, polylysine, poly(acrylonitrile-co-acrylic acidacrylamidine), polyacrylic acid, polyhydroxyethylmethacrylate, polyvinylpyrrolidone, polyethylene glycol, alginate, sepharose, agarose, polyethyleneimine, polyallylamine, polyaminoethylmethacrylate, polyornithine, polyaminoacrylamide, polyacrylamide, polyacrolein, polyacryloxysuccinimide, polysulfone, poly(acrylonitrile-co-acrylamide), and their copolymers, either alone or in combination.

27. The material of claim 17 wherein the hydrophilic polymer comprises a random copolymer of tetrafluoroethylene and vinyl alcohol.

28. The material of claim 17 wherein the hydrophilic polymer comprises a copolymer of hydroxyethylmethacrylate and hexafluoroacrylate.

29. The material of claim 17 wherein the hydrophilic polymer comprises a copolymer of polyethylene and vinyl alcohol.

30. The material of claim 17 further comprising a spacer compound for interposition between the second layer and a bioactive species.

31. The material of claim 30 wherein the spacer compound is selected from a member of a group consisting of succinic acid, diaminohexane, glyoxylic acid, short chain polyethylene glycol, and glycine.