Surface functionalized Langmuir-Blodgett films for immobilization of active moieties

• US 5,102,798 A
• Filed: 09/08/1988
• Issued: 04/07/1992
• Est. Priority Date: 09/08/1988
• Status: Expired due to Fees

• Current Assignee: Allage Associates
• Original Assignee: Allage Associates
• Inventors: Guiseppi-Elie, Anthony
• Primary Examiner(s): Naff, David M.
• Application Number:
  US07/241,607
• Time in Patent Office: 1,307 Days
• Field of Search: 435/174, 435/177, 435/180, 435/181, 436/528, 436/531, 436/532, 530/812, 530/815, 530/816, 264/165
• US Class Current: 435/177
• CPC Class Codes: C12N 11/02 Enzymes or microbial cells immobilised on or in an organic carrier
• CPC Class Codes: C12N 11/082 obtained by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: G01N 33/5432 Liposomes or microcapsules
• CPC Class Codes: Y10S 530/812 Peptides or proteins is immobilized on, or in, an organic carrier
• CPC Class Codes: Y10S 530/815 Carrier is a synthetic polymer
• CPC Class Codes: Y10S 530/816 Attached to the carrier via a bridging agent

Abstract

Langmuir-Blodgett films having hydrophobic surfaces are chemically modified to convert the hydrophobic surfaces to hydrophilic surfaces for immobilization of active moieties having bioactive, immunoactive, thermoactive, electroactive, optoactive or redoxactive properties. Langmuir-Blodgett films having a hydrophobic surface of omega unsaturated covalent bonds are formed from an amphiphilic, bifunctional surface active material having a hydrophobic tail group bearing an omega terminus of double or triple bonded unsaturation. The amphiphilic material may be fatty acids, phospholipids or porphyrins, and is preferably omega-tricosenoylamide or omega-tricosenoic acid. Chemical modification is carried out by exposing the hydrophobic surface to a reagent such as alkaline or acidic potassium permanganate, potassium dichromate or osmium tetroxide which oxidizes the omega unsaturated covalent bonds to provide hydrophilic groups such as hydroxyl or carboxylic acid groups. These groups make the film surface hydrophilic and function as sites for attachment of the active moieties.

Claims

1. A process for chemically modifying a hydrophobic surface of a Langmuir-Blodgett film so as to make said surface hydrophilic, said process comprising:

a. providing a Langmuir-Blodgett film having a hydrophobic surface of omega unsaturated covalent bonds formed from an amphiphilic, bifunctional surface active material having a hydrophobic tail group bearing an omega terminus of double or triple bonded unsaturation; and

b. exposing the hydrophobic surface of the Langmuir-Blodgett film to a reagent which reacts with the surface of the Langmuir-Blodgett film for a time sufficient so as to oxidize said omega unsaturated covalent bonds to provide hydrophilic groups on said surface thus providing a hydrophilic surface on the Langmuir-Blodgett film.

2. The process of claim 1 wherein the amphiphilic material is selected from the group consisting of bifunctional, surface active molecules having both ionizable and non-ionizable head groups.

3. The process of claim 2 wherein the amphiphilic material is selected from the group consisting of fatty acids, phospholipids, porphyrins and their derivatives.

4. The process of claim 3 wherein the fatty acids are selected from the group consisting of arachidic acid, stearic acid, heptadecanoic acid, hexadeconic acid, and their derivatives.

5. The process of claim 3 wherein the porphyrins are selected from the group consisting of phthalocyanine, merocyanine and their derivatives.

6. The process of claim 1 wherein the amphiphilic material is selected from the group consisting of omega-tricosenoylamide and omega-tricosenoic acid.

7. The process of claim 1 wherein the reagent which reacts with the surface provides a surface having a hydroxyl functionality.

8. The process of claim 7 wherein the reagent which reacts with the surface is an aqueous reagent selected from the group consisting of alkaline potassium permanganate, potassium dichromate and osmium tetroxide.

9. The process of claim 1 wherein the reagent which reacts with the surface provides a surface having a carboxylic acid functionality.

10. The process of claim 9 wherein the reagent which reacts with the surface is an aqueous reagent comprising acidic potassium permanganate.

11. The process of claim 1 comprising the additional step of exposing the surface of the Langmuir-Blodgett film to additional chemical reagents subsequent to providing the hydrophilic groups, said additional reagents being reactive with the hydrophilic groups on the surface of the Langmuir-Blodgett film, and said exposure being for a time sufficient to immobilize the additional chemical reagents on the surface of the Langmuir-Blodgett film.

12. process of claim 11 wherein the additional chemical reagent is selected from the group consisting of biologically active reagents.

13. The process of claim 12 wherein the biologically active reagents are selected from the group consisting of enzymes, enzyme conjugates, cofactors, antigens, antibodies, RNA, DNA, hormones, enzyme inhibitors and enzyme substrates.

14. The process of claim 13 wherein the biologically active reagent comprises glucose oxidase.

15. The process of claim 13 wherein the biologically active reagent comprises flavin adenine dinucleotide.

16. The process of claim 1 comprising the additional step of depositing a subsequent Langmuir-Blodgett film upon the Langmuir-Blodgett film containing hydrophilic groups.

17. The process of claim 16 wherein the subsequent Langmuir-Blodgett film is of a composition different from that of the Langmuir-Blodgett film containing hydrophilic groups.

18. The process of claim 16 wherein the subsequent Langmuir-Blodgett film comprises built-up monolayers of amphiphilic material.

19. The process of claim 18 wherein the amphiphilic material is selected from the group consisting of bifunctional, surface active molecules having both ionizable and non-ionizable head groups.

20. The process of claim 19 wherein the amphiphilic material is selected from the group consisting of fatty acids, phospholipids, porphyrins and their derivatives.

21. The process of claim 20 wherein the fatty acids are selected from the group consisting of arachidic acid, stearic acid, heptadecanoic acid, hexadeconic acid, and their derivatives.

22. The process of claim 20 wherein the porphyrins are selected from the group consisting of phthalocyanine, merocyanine and their derivatives.

23. The process of claim 18 wherein the amphiphilic material is selected from the group consisting of bifunctional, surface active molecules having hydrophobic tail groups bearing an omega terminus of double or triple bonded unsaturation.

24. The process of claim 23 wherein the amphiphilic material is selected from the group consisting of omega-tricosenoylamide and omega-tricosenoic acid.

25. The process of claim 3 wherein the phospholipids are selected from the group consisting of phosphatidylcholine, bis-[12-(methacryloyloxy)dodecanoyl]-L-phosphatidylcholine and their derivatives.

26. The process of claim 20 wherein the phospholipids are selected from the group consisting of phsophatidylcholine, bis-[12-(methacryloyloxy)dodecanoyl]-L-phosphatidylcholine and their derivatives.

27. A Langmuir-Blodgett film having a hydrophilic surface prepared by the process of claim 1.

28. A process for preparing a Langmuir-Blodgett film/supra-molecular complex, said process comprising:

a. providing a Langmuir-Blodgett film having a hydrophobic surface of omega unsaturated covalent bonds formed from an amphiphilic, bifunctional surface active material having a hydrophobic tail group bearing an omega terminus of double or triple bonded unsaturation;

b. exposing the hydrophobic surface of the Langmuir-Blodgett film to a reagent which reacts with the surface of the Langmuir-Blodgett film for a time sufficient so as to oxidize said omega unsaturated covalent bonds on said surface so as to form hydroxyl or carboxylic acid groups thus providing a hydrophilic surface on the Langmuir-Blodgett film; and

c. exposing the hydrophilic surface of the Langmuir-Blodgett film to a chemical reagent that is active to the surface of the film for a time sufficient to provide immobilization of the chemical reagent on the film surface, to thereby result in the creation of a Langmuir-Blodgett film/supra-molecular complex.

29. A Langmuir-Blodgett film/supra-molecular complex prepared by the following steps:

a. providing a Langmuir-Blodgett film having a hydrophobic surface of omega unsaturated covalent bonds formed from an amphiphilic, bifunctional surface active material having a hydrophobic tail group bearing an omega terminus of double or triple bonded unsaturation;

b. exposing the hydrophobic surface of said Langmuir-Blodgett film to reagents which react with the surface of the Langmuir-Blodgett film for a time sufficient so as to oxidize said omega unsaturated covalent bonds so as to form hydroxyl or carboxylic acid groups on said surface thus providing a hydrophilic surface on the Langmuir-Blodgett film; and

c. exposing the hydrophilic surface of the Langmuir-Blodgett film to an additional chemical reagent that is active to the surface of the film for a time sufficient to provide immobilization of the additional chemical reagent on the film surface to result in a Langmuir-Blodgett film/supra-molecular complex.

30. The complex of claim 29 wherein built-up monolayers of amphiphilic material are formed on the Langmuir-Blodgett film having a hydrophilic surface.

31. The complex of claim 30 wherein the amphiphilic material is selected from the group consisting of bifunctional, surface active molecules having both ionizable and non-ionizable head groups.

32. The complex of claim 31 wherein the amphiphilic material is selected from the group consisting of fatty acids, phospholipids, porphyrins and their derivatives.

33. The complex of claim 32 wherein the fatty acids are selected from the group consisting of arachidic acid, stearic acid, heptadecanoic acid, hexadeconic acid, and their derivatives.

34. The complex of claim 32 wherein the porphyrins are selected from the group consisting of phthalocyanine, merocyanine and their derivatives.

35. The complex of claim 30 wherein the amphiphilic material is selected from the group consisting of bifunctional, surface active molecules having hydrophobic tail groups bearing an omega terminus of double or triple bonded unsaturation.

36. The complex of claim 30 wherein the amphiphilic material is selected from the group consisting of omega-tricosenoylamide and omega-tricosenoic acid.

37. The complex of claim 29 wherein the reagent which reacts with the surface provides a surface having a hydroxyl functionality.

38. The complex of claim 31 wherein the reagent which reacts with the surface provides a surface having a carboxyl functionality.

39. The complex of claim 29 wherein the additional chemical reagent is selected from the group consisting of biologically active reagents.

40. The complex of claim 39 wherein the biologically active reagents are selected from the group consisting of enzymes, cofactors, antigens, antibodies, enzyme conjugates, RNA, DNA, hormones, enzyme inhibitors and enzyme substrates.

41. The complex of claim 40 wherein the biologically active reagent comprises glucose oxidase.

42. The complex of claim 40 wherein the biologically active reagent comprises flavin adenine dinucleotide.

43. The complex of claim 29 wherein the additional chemical reagent comprises a subsequent Langmuir-Blodgett film.

44. The complex of claim 43 wherein the subsequent Langmuir-Blodgett film is of a composition different from that of the precursor Langmuir-Blodgett film.

45. The complex of claim 44 wherein the subsequent Langmuir-Blodgett film comprises octadecylamine.

46. The complex of claim 32 wherein the phospholipids are selected from the group consisting of phsophatidylcholine, bis-[12-(methacryloyloxy)-dodecanoyl]-L-phosphatidylcholine and their derivatives.

47. A process for chemically modifying a hydrophobic surface of a Langmuir-Blodgett film so as to provide a hydroxyl or carboxylic acid functionality on said surface, said process comprising:

a. providing a Langmuir-Blodgett film having a hydrophobic surface of omega unsaturated covalent bonds formed from an amphiphilic, bifunctional surface active material having a hydrophobic tail group bearing an omega terminus of double or triple bonded unsaturation; and

,

b. exposing the hydrophobic surface of the Langmuir-Blodgett film to a reagent which reacts with the surface of the Langmuir-Blodgett film for a time sufficient so as to oxidize said omega unsaturated covalent bonds on the surface thus providing a carboxylic acid or hydroxyl functionality on the Langmuir-Blodgett film.

48. A process for chemically modifying a hydrophobic surface of a Langmuir-Blodgett film so as to make said surface hydrophilic, said process comprising:

a. providing a Langmuir-Blodgett film having a hydrophobic surface of omega unsaturated covalent bonds, said Langmuir-Blodgett film comprising built-up monolayers of amphilphilic material wherein the amphiphilic material is selected from the group consisting of omega-tricosenoylamide and omega-tricosenoic acid; and

b. exposing the hydrophobic surface of the Langmuir-Blodgett film to a regent which reacts with the surface of the Langmuir-Blodgett film for a time sufficient so as to oxidize said omega unsaturated covalent bonds to provide hydrophilic groups on said surface thus providing a hydrophilic surface on the Langmuir-Blodgett film.