Patterning antibodies on a surface

• US 5,858,801 A
• Filed: 03/13/1997
• Issued: 01/12/1999
• Est. Priority Date: 03/13/1997
• Status: Expired due to Term

• Current Assignee: the united states of america as represented by the secretary of the navy
• Original Assignee: the united states of america as represented by the secretary of the navy
• Inventors: Brizzolara, Robert A.
• Primary Examiner(s): Chin, Christopher L.
• Assistant Examiner(s): NGUYEN, BAO THUY L
• Application Number:
  US08/816,337
• Time in Patent Office: 670 Days
• Field of Search: 430/302, 430/304, 430/298, 430/297, 430/299, 422/57, 422/60, 435/33, 435/287.8, 435/287.9, 435/973, 436/518, 436/527, 427/2.11, 427/2.13, 427/466, 427/470, 427/504, 427/534, 427/288, 427/286
• US Class Current: 436/518
• CPC Class Codes: G01N 33/54386 Analytical elements
• CPC Class Codes: Y10S 435/973 Simultaneous determination of more than one analyte
• CPC Class Codes: Y10S 977/856 including etching/cutting

Abstract

A patterned multiple antibody substrate for use in biosensors or immunosensors is produced by (1) coating an antibody-adsorbent substrate with a material that resists antibody adsorption, (2) using ion beam sputtering, laser ablation, or mechanical scribing to remove the coating at specific sites on the substrate, and then (3) adsorbing specific antibodies at the sites. The substrate is capable of detecting multiple chemical species simultaneously.

Claims

1. A process for producing a multiple antibody patterned substrate comprising:

A. coating an antibody-adsorbent substrate with an antibody-resistant material that is resistant to adsorption of antibodies;

B. removing a portion of the antibody-resistant material to produce a bare site on the antibody-adsorbent substrate having a precise size, shape, and location on the substrate;

C. adsorbing molecules of a selected antibody on to the bare site on the antibody adsorbent-substrate;

D. rinsing the substrate to remove unadsorbed antibody molecules;

E. coating the antibody-adsorbent substrate with more of the antibody-resistant material to cover the bare surface of the substrate between the newly adsorbed antibody molecules; and

F. repeating steps B through E until each of the antibodies has been adsorbed at its specific site on the antibody adsorbent substrate.

2. The process of claim 1 wherein the antibody-adsorbent substrate is composed of a hydrophobic material.

3. The process of claim 2 wherein the antibody-adsorbent substrate is composed of a hydrophobic, polymeric material.

4. The process of claim 3 wherein the antibody-adsorbent substrate is composed of polystyrene, polypropylene, polyethylene, or copolymers of polypropylene and polyethylene.

5. The process of claim 4 wherein the antibody-adsorbent substrate is composed of polystyrene or polypropylene.

6. The process of claim 1 wherein the antibody-resistant material is bovine serum albumin, gelatin, lysozyme, octoxynol, polyoxyethylenesorbitan monolaurate, or a polyethylene oxide containing block copolymer surfactant.

7. The process of claim 6 wherein the antibody-resistant material is bovine serum albumin, gelatin, or lysozyme.

8. The process of claim 7 wherein the antibody-resistant material is bovine serum albumin.

9. A process for producing a multiple antibody patterned substrate comprising:

A. coating an antibody-adsorbent substrate with an antibody-resistant material that is resistant to the adsorption of antibodies;

B. removing portions of the antibody-resistant material to produce bare sites on the antibody-adsorbent substrate having precise sizes and shapes and each site having a precise location which corresponds to a specific antibody;

C. adsorbing molecules of each antibody to its specific bare site on the antibody-adsorbent substrate;

D. rinsing the substrate to remove unadsorbed antibodies; and

E. coating the antibody-adsorbent substrate with more of the antibody-resistant material to cover the bare surface of the substrate between the adsorbed antibody molecules.

10. The process of claim 9 wherein each antibody adsorbed in step C is delivered to its precise bare site on the antibody-adsorbent substrate by micropipetting.

11. The process of claim 9 wherein each antibody adsorbed in step C is delivered to its precise bare site on the antibody-absorbent substrate by an ink jet printer.

12. The process of claim 9 wherein the antibody-adsorbent substrate is composed of a hydrophobic material.

13. The process of claim 12 wherein the antibody-adsorbent substrate is composed of a hydrophobic, polymeric material.

14. The process of claim 13 wherein the antibody adsorbent substrate is composed of polystyrene, polypropylene, polyethylene, or copolymers of polypropylene and polyethylene.

15. The process of claim 14 wherein the antibody-adsorbent substrate is composed of polystyrene or polypropylene.

16. The process of claim 9 wherein the antibody-resistant material is bovine serum albumin, gelatin, lysozyme, octoxynol, polyoxyethylenesorbitan monolaurate, or a polyethylene oxide containing block copolymer surfactant.

17. The process of claim 16 wherein the antibody-resistant material is bovine serum albumin, gelatin, or lysozyme.

18. The process of claim 17 wherein the antibody-resistant material is bovine serum albumin.

19. A process for producing a multiple antibody patterned substrate comprising:

A. coating an antibody-adsorbent substrate with an antibody-resistant material that is resistant to adsorption of antibodies;

B. removing a portion of the antibody-resistant material by ion beam sputtering to produce a bare site on the antibody-adsorbent substrate having a precise size, shape, and location on the substrate;

C. adsorbing molecules of a selected antibody on to the bare site on the antibody adsorbent-substrate;

D. rinsing the substrate to remove unadsorbed antibody molecules;

E. coating the antibody-adsorbent substrate with more of the antibody-resistant material to cover the bare surface of the substrate between the newly adsorbed antibody molecules; and

F. repeating steps B through E until each of the antibodies has been adsorbed at its specific site on the antibody adsorbent substrate.

20. The process of claim 19 wherein the antibody-adsorbent substrate is composed of a hydrophobic material.

21. The process of claim 20 wherein the antibody-adsorbent substrate is composed of a hydrophobic, polymeric material.

22. The process of claim 21 wherein the antibody-adsorbent substrate is composed of polystyrene, polypropylene, polyethylene, or copolymers of polypropylene and polyethylene.

23. The process of claim 22 wherein the antibody-adsorbent substrate is composed of polystyrene or polypropylene.

24. The process of claim 19 wherein the antibody-resistant material is bovine serum albumin, gelatin, lysozyme, octoxynol, polyoxyethylenesorbitan monolaurate, or a polyethylene oxide containing block copolymer surfactant.

25. The process of claim 24 wherein the antibody-resistant material is bovine serum albumin, gelatin, or lysozyme.

26. The process of claim 25 wherein the antibody-resistant material is bovine serum albumin.

27. A process for producing a multiple antibody patterned substrate comprising:

A. coating an antibody-adsorbent substrate with an antibody-resistant material that is resistant to adsorption of antibodies;

B. removing a portion of the antibody-resistant material by laser ablation to produce a bare site on the antibody-adsorbent substrate having a precise size, shape, and location on the substrate;

C. adsorbing molecules of a selected antibody on to the bare site on the antibody adsorbent-substrate;

D. rinsing the substrate to remove unadsorbed antibody molecules;

E. coating the antibody-adsorbent substrate with more of the antibody-resistant material to cover the bare surface of the substrate between the newly adsorbed antibody molecules; and

F. repeating steps B through E until each of the antibodies has been adsorbed at its specific site on the antibody adsorbent substrate.

28. The process of claim 27 wherein the antibody-adsorbent substrate is composed of a hydrophobic material.

29. The process of claim 28 wherein the antibody-adsorbent substrate is composed of a hydrophobic, polymeric material.

30. The process of claim 29 wherein the antibody-adsorbent substrate is composed of polystyrene, polypropylene, polyethylene, or copolymers of polypropylene and polyethylene.

31. The process of claim 30 wherein the antibody-adsorbent substrate is composed of polystyrene or polypropylene.

32. The process of claim 27 wherein the antibody-resistant material is bovine serum albumin, gelatin, lysozyme, octoxynol, polyoxyethylenesorbitan monolaurate, or a polyethylene oxide containing block copolymer surfactant.

33. The process of claim 32 wherein the antibody-resistant material is bovine serum albumin, gelatin, or lysozyme.

34. The process of claim 33 wherein the antibody-resistant material is bovine serum albumin.

35. A process for producing a multiple antibody patterned substrate comprising:

A. coating an antibody-adsorbent substrate with an antibody-resistant material that is resistant to adsorption of antibodies;

B. removing a portion of the antibody-resistant material by laser mechanical scribing using atomic force microscopy to produce a bare site on the antibody-adsorbent substrate having a precise size, shape, and location on the substrate;

C. adsorbing molecules of a selected antibody on to the bare site on the antibody adsorbent-substrate;

D. rinsing the substrate to remove unadsorbed antibody molecules;

E. coating the antibody-adsorbent substrate with more of the antibody-resistant material to cover the bare surface of the substrate between the newly adsorbed antibody molecules; and

F. repeating steps B through E until each of the antibodies has been adsorbed at its specific site on the antibody adsorbent substrate.

36. The process of claim 35 wherein the antibody-adsorbent substrate is composed of a hydrophobic material.

37. The process of claim 36 wherein the antibody-adsorbent substrate is composed of a hydrophobic, polymeric material.

38. The process of claim 37 wherein the antibody-adsorbent substrate is composed of polystyrene, polypropylene, polyethylene, or copolymers of polypropylene and polyethylene.

39. The process of claim 38 wherein the antibody-adsorbent substrate is composed of polystyrene or polypropylene.

40. The process of claim 35 wherein the antibody-resistant material is bovine serum albumin, gelatin, lysozyme, octoxynol, polyoxyethylenesorbitan monolaurate, or a polyethylene oxide containing block copolymer surfactant.

41. The process of claim 40 wherein the antibody-resistant material is bovine serum albumin, gelatin, or lysozyme.

42. The process of claim 41 wherein the antibody-resistant material is bovine serum albumin.

43. A process for producing a multiple antibody patterned substrate comprising:

A. coating an antibody-adsorbent substrate with an antibody-resistant material that is resistant to the adsorption of antibodies;

B. removing portions of the antibody-resistant material by ion beam sputtering to produce bare sites on the antibody-adsorbent substrate having precise sizes and shapes and each site having a precise location which corresponds to a specific antibody;

C. adsorbing molecules of each antibody to its specific bare site on the antibody-adsorbent substrate;

D. rinsing the substrate to remove unadsorbed antibodies; and

E. coating the antibody-adsorbent substrate with more of the antibody-resistant material to cover the bare surface of the substrate between the adsorbed antibody molecules.

44. The process of claim 43 wherein each antibody adsorbed in step C is delivered to its precise bare site on the antibody-adsorbent substrate by micropipetting.

45. The process of claim 43 wherein each antibody adsorbed in step C is delivered to its precise bare site on the antibody-absorbent substrate by an ink jet printer.

46. The process of claim 43 wherein the antibody-adsorbent substrate is composed of a hydrophobic material.

47. The process of claim 46 wherein the antibody-adsorbent substrate is composed of a hydrophobic, polymeric material.

48. The process of claim 47 wherein the antibody adsorbent substrate is composed of polystyrene, polypropylene, polyethylene, or copolymers of polypropylene and polyethylene.

49. The process of claim 48 wherein the antibody-adsorbent substrate is composed of polystyrene or polypropylene.

50. The process of claim 43 wherein the antibody-resistant material is bovine serum albumin, gelatin, lysozyme, octoxynol, polyoxyethylenesorbitan monolaurate, or a polyethylene oxide containing block copolymer surfactant.

51. The process of claim 50 wherein the antibody-resistant material is bovine serum albumin, gelatin, or lysozyme.

52. The process of claim 51 wherein the antibody-resistant material is bovine serum albumin.

53. A process for producing a multiple antibody patterned substrate comprising:

A. coating an antibody-adsorbent substrate with an antibody-resistant material that is resistant to the adsorption of antibodies;

B. removing portions of the antibody-resistant material by laser ablation to produce bare sites on the antibody-adsorbent substrate having precise sizes and shapes and each site having a precise location which corresponds to a specific antibody;

C. adsorbing molecules of each antibody to its specific bare site on the antibody-adsorbent substrate;

D. rinsing the substrate to remove unadsorbed antibodies; and

E. coating the antibody-adsorbent substrate with more of the antibody-resistant material to cover the bare surface of the substrate between the adsorbed antibody molecules.

54. The process of claim 53 wherein each antibody adsorbed in step C is delivered to its precise bare site on the antibody-adsorbent substrate by micropipetting.

55. The process of claim 53 wherein each antibody adsorbed in step C is delivered to its precise bare site on the antibody-absorbent substrate by an ink jet printer.

56. The process of claim 53 wherein the antibody-adsorbent substrate is composed of a hydrophobic material.

57. The process of claim 56 wherein the antibody-adsorbent substrate is composed of a hydrophobic, polymeric material.

58. The process of claim 57 wherein the antibody adsorbent substrate is composed of polystyrene, polypropylene, polyethylene, or copolymers of polypropylene and polyethylene.

59. The process of claim 58 wherein the antibody-adsorbent substrate is composed of polystyrene or polypropylene.

60. The process of claim 53 wherein the antibody-resistant material is bovine serum albumin, gelatin, lysozyme, octoxynol, polyoxyethylenesorbitan monolaurate, or a polyethylene oxide containing block copolymer surfactant.

61. The process of claim 60 wherein the antibody-resistant material is bovine serum albumin, gelatin, or lysozyme.

62. The process of claim 61 wherein the antibody-resistant material is bovine serum albumin.

63. A process for producing a multiple antibody patterned substrate comprising:

A. coating an antibody-adsorbent substrate with an antibody-resistant material that is resistant to the adsorption of antibodies;

B. removing portions of the antibody-resistant material by mechanical scribing using atomic force microscopy to produce bare sites on the antibody-adsorbent substrate having precise sizes and shapes and each site having a precise location which corresponds to a specific antibody;

C. adsorbing molecules of each antibody to its specific bare site on the antibody-adsorbent substrate;

D. rinsing the substrate to remove unadsorbed antibodies; and

E. coating the antibody-adsorbent substrate with more of the antibody-resistant material to cover the bare surface of the substrate between the adsorbed antibody molecules.

64. The process of claim 63 wherein each antibody adsorbed in step C is delivered to its precise bare site on the antibody-adsorbent substrate by micropipetting.

65. The process of claim 63 wherein each antibody adsorbed in step C is delivered to its precise bare site on the antibody-absorbent substrate by an ink jet printer.

66. The process of claim 63 wherein the antibody-adsorbent substrate is composed of a hydrophobic material.

67. The process of claim 66 wherein the antibody-adsorbent substrate is composed of a hydrophobic, polymeric material.

68. The process of claim 67 wherein the antibody adsorbent substrate is composed of polystyrene, polypropylene, polyethylene, or copolymers of polypropylene and polyethylene.

69. The process of claim 68 wherein the antibody-adsorbent substrate is composed of polystyrene or polypropylene.

70. The process of claim 63 wherein the antibody-resistant material is bovine serum albumin, gelatin, lysozyme, octoxynol, polyoxyethylenesorbitan monolaurate, or a polyethylene oxide containing block copolymer surfactant.

71. The process of claim 70 wherein the antibody-resistant material is bovine serum albumin, gelatin, or lysozyme.

72. The process of claim 71 wherein the antibody-resistant material is bovine serum albumin.