Method for attachment of biomolecules to surfaces through amine-functional groups

• US 6,303,179 B1
• Filed: 02/08/1999
• Issued: 10/16/2001
• Est. Priority Date: 02/08/1999
• Status: Expired due to Fees

• Current Assignee: Medtronic Incorporated (Medtronic Plc)
• Original Assignee: Medtronic Incorporated (Medtronic Plc)
• Inventors: Vincent, Judith, Verhoeven, Michel, Cahalan, Linda, Cahalan, Patrick, Koulik, Edouard
• Primary Examiner(s): Nguyen, Dinh X.
• Assistant Examiner(s): BARRETT, THOMAS C
• Application Number:
  US09/245,834
• Time in Patent Office: 981 Days
• Field of Search: 427/2--, 564/488, 564/414, 525/50, 623/11.11
• US Class Current: 427/2.26
• CPC Class Codes: A61L 33/0029 using an intermediate layer of polymer

Abstract

A method of modifying the surface characteristics of a substrate having a surface with an amide-functional polymer thereon. The method involves contacting the amide-functional polymer with a source of hydroxide ions and a source of hypohalite ions at a temperature of at least about 20° C. for a time effective to convert at least a portion of the amide-functional groups to amine-functional groups to form a substrate surface comprising an amine-functional polymer, wherein the hydroxide ions are present in a molar excess relative to the hypohalite ions and at a concentration of no more than about 0.1 M, based on the total volume of the reaction mixture. A biomolecule is attached to the resultant amine-functional polymer.

Claims

1. A method of making a medical device having a biomolecule immobilized on a substrate surface, the method comprising:

providing the substrate, the substrate having a surface comprising an amide-functional polymer;

contacting the amide-functional polymer with a reaction mixture comprising a source of hydroxide ions and a source of hypohalite ions at a temperature of at least about 20°

C. for a period of time effective to convert at least a portion of the amide-functional groups to amine-functional groups to form an amine-functional polymer on the substrate surface, the period of time not exceeding about three hours, wherein the hydroxide ions are present in a molar excess relative to the hypohalite ions, and wherein the hydroxide ions are present at a concentration of no more than about 0.1 M, based on the total volume of the reaction mixture; and

contacting the substrate surface comprising an amine-functional polymer with a biomolecule under conditions effective to immobilize the biomolecule on the substrate surface.

2. The method of claim 1 wherein the step of contacting the amide-functional polymer comprises immersing the substrate in the reaction mixture.

3. The method of claim 1 wherein the source of hydroxide ions comprises one or more hydroxide bases and the source of hypohalite ions comprises one or more hypohalous acids, one or more hypohalite salts, or mixtures thereof.

4. The method of claim 1 wherein the ratio of moles of hydroxide ions to moles of hypohalite ions is at least about 3:

1.

5. The method of claim 4 wherein the ratio of moles of hydroxide ions to moles of hypohalite ions is no more than about 20:

1.

6. The method of claim 1 wherein the reaction mixture comprises one or more hydroxide bases at a total concentration of at least about 0.01 M hydroxide ions, based on the total volume of the reaction mixture, and one or more hypohalous acids, one or more hypohalite salts, or mixtures thereof, at a total concentration of at least about 0.001 M to about 0.03 M hypohalite ions, based on the total volume of the reaction mixture.

7. The method of claim 1 wherein the step of contacting the amide-functional polymer comprises contacting it with a reaction mixture comprising a source of hydroxide ions and a source of hypohalite ions in water at a temperature of about 20°

C. to about 30°

C. for at least about 15 minutes.

8. The method of claim 1 wherein the temperature is within a range of about 20°

C. to about 30°

C.

9. The method of claim 1 wherein the biomolecule is selected from the group of an antibacterial agent, an antimicrobial agent, an anticoagulant, an antithrombotic agent, an antiplatelet agent, an anti-inflammatory, an enzyme, a catalyst, a hormone, a growth factor, a drugs, a vitamin, an antibody, an antigen, a nucleic acid, a dye, a DNA, an RNA, a protein, a peptide, and mixtures thereof.

10. The method of claim 9 wherein the biomolecule is synthetically derived or naturally occurring.

11. The method of claim 9 wherein the biomolecule comprises heparin or a salt thereof.

12. The method of claim 1 wherein the substrate comprises a solid organic polymer or a metal.

13. The method of claim 12 wherein the substrate comprises a solid organic polymer selected from the group of a polyurethane, a polyolefin, a silicone, a fluoropolymer, a polyester, a polyvinyl halide, a polyether, a polyamide, and blends and copolymers thereof.

14. The method of claim 13 wherein the substrate comprises a polyurethane or a polyvinyl chloride.

15. The method of claim 13 wherein the substrate comprises a solid organic polymer surface having an amide-functional polymer grafted thereto.

16. The method of claim 1 wherein the substrate comprises a metal surface coated with a vinylsilane having an amide-functional polymer grafted thereto.

17. The method of claim 1 wherein the source of hydroxide ions comprises sodium hydroxide, potassium hydroxide, or a mixture thereof.

18. The method of claim 1 wherein the source of hypohalite ions comprises HOCl, LiOCl, NaOCl, KOCl, HOBr, LiOBr, NaOBr, KOBr, HOl, LiOl, NaOl, KOl, Ca(OCl)₂, Sr(OCl)₂, Ba(OCl)₂ Ca(OBr)₂, Sr(OBr)₂, Ba(OBr)₂, Ca(Ol)₂, Sr(Ol)₂, Ba(Ol)₂, or mixtures thereof.

19. The method of claim 1 wherein the surface formed is biocompatible.

28. A medical device preparable by the method of claim 1.

20. A method of modifying the surface characteristics of a solid polymeric material, the method comprising:

irradiating a surface of the solid polymeric material;

contacting the irradiated surface with an amide-functional ethylenically unsaturated monomer and a source of oxidizing metal ions under conditions effective to graft the monomer to the irradiated surface to form an amide-functional graft polymer thereon;

contacting the amide-functional graft polymer with a reaction mixture comprising a source of hydroxide ions and a source of hypohalite ions at a temperature of at least about 20°

C. for a period of time effective to convert at least a portion of the amide-functional groups to amine-functional groups to form a solid polymeric material having a surface with an amine-functional graft polymer thereon, the period of time not exceeding about three hours, wherein the hydroxide ions are present in a molar excess relative to the hypohalite ions, and wherein the hydroxide ions are present at a concentration of no more than about 0.1 M, based on the total volume of the reaction mixture; and

contacting the solid polymeric material having a surface with an amine-functional graft polymer thereon with a biomolecule under conditions effective to immobilize the biomolecule on the surface.

21. The method of claim 20 wherein the source of hydroxide ions comprises one or more hydroxide bases and the source of hypohalite ions comprises one or more hypohalous acids, one or more hypohalite salts, or mixtures thereof.

22. The method of claim 20 wherein the ratio of moles of hydroxide ions to moles of hypohalite ions is at least about 3:

1 and no more than about 20;

1.

23. The method of claim 20 wherein the temperature is within a range of about 20°

C. to about 30°

C.

24. The method of claim 20 wherein the biomolecule is heparin or a salt thereof and the ethylenically unsaturated monomer is acrylamide.

25. The method of claim 20 wherein the surface is modified with up to about 50 μ

g of the biomolecule per square centimeter.

26. The method of claim 20 wherein the surface formed is biocompatible.

27. A modified polymeric material preparable by the method of claim 20.