Method and apparatus for the detection of an enzyme

• US 20060035307A1
• Filed: 08/13/2004
• Published: 02/16/2006
• Est. Priority Date: 08/13/2004
• Status: Active Grant

• Current Assignee: Biomed Solutions, LLC
• Original Assignee: Biomed Solutions, LLC
• Inventors: Bucholtz, Kevin, Horner, Scott, Taghizadeh, Farhan
• Granted Patent: US 7,402,423 B2
• Time in Patent Office: Days
• US Class Current: 435/23
• CPC Class Codes: C12Q 1/37 involving peptidase or proteinase

Abstract

An apparatus for detecting pepsin comprising a solid support, and a peptide chain wherein the peptide chain is operatively configured to be cleaved by pepsin, and the peptide chain is disposed on a surface of the solid support.

Claims

1. An apparatus for detecting an enzyme comprising a test strip, and non-antibody means for detecting a protein that has been cleaved by an enzyme.

2. The apparatus as recited in claim 1, wherein said enzyme is pepsin.

3. An apparatus for detecting an enzyme comprising

a. a solid support, and

b. a peptide chain wherein

i. said peptide chain is cleavable by an enzyme,

ii. said peptide chain is disposed on a surface of said solid support.

4. The apparatus as recited in claim 3, wherein said enzyme is pepsin.

5. An apparatus for detecting an enzyme comprising

a. a solid support, and

b. a peptide chain comprised of a first end and a second end wherein

i. said first end of said peptide chain is attached to a surface of said solid support,

ii. said second end of said peptide chain is attached to a first tag, thereby attaching said first tag to said solid support, wherein said peptide chain is cleavable by an enzyme, such that, when said peptide chain is cleaved by said enzyme, said first tag is released from said solid support to generate a released tag.

6. The apparatus as recited in claim 5, wherein said enzyme comprises pepsin.

7. The apparatus as recited in claim 6, wherein said first tag comprises a functional group that reacts with a diazonium compound to produce an azo compound.

8. The apparatus as recited in claim 6, wherein said functional group is an aromatic alcohol.

9. The apparatus as recited in claim 8, wherein said aromatic alcohol is a substituted naphthol.

10. The apparatus as recited in claim 8, wherein said aromatic alcohol is a substituted phenol.

11. A process for detecting an enzyme comprising the steps of

a. exposing a first solution to a test strip to generate a treated strip wherein said test strip comprises

i. a solid support, and

ii. a peptide chain wherein said peptide chain is cleavable by pepsin, thereby generating a released particle and a bound particle,

b. exposing said treated strip to a second solution wherein said second solution comprises a chemical which reacts with a detected particle selected from the group consisting of said released particle, said bound particle, and combinations thereof.

12. The process as recited in claim 11 wherein said first solution comprises pepsin.

13. The process as recited in claim 12 wherein said released particle is a first tag.

14. The process as recited in claim 13 wherein said first tag is an aromatic alcohol.

15. The process as recited in claim 12 wherein said released particle is a released fragment of said peptide chain.

16. The process as recited in claim 15, wherein said chemical is Coumassie Blue.

17. The process as recited in claim 12, wherein said chemical is a second tag.

18. The process as recited in claim 17, wherein said second tag is an azo compound.

19. The process as recited in claim 18, wherein said azo compound is an aromatic azo compound.

20. A process for detecting an enzyme comprising the steps of:

a. obtaining a sample to be tested from a patient,

b. expositing said sample to a testing substrate comprised of means for detecting pepsin,

c. detecting a presence of pepsin in said sample with said means for detecting pepsin, wherein time elapsed between the obtaining of said sample and the detecting of said presence of pepsin is less than about 2 hours.

21. The process as recited in claim 20, wherein said means for detecting pepsin is a non-antibody means.

22. The process as recited in claim 20, wherein said testing substrate is further comprised of a solid support.

23. The process as recited in claim 22, wherein said time elapsed is less than about 1 hour.

24. An article for detecting the presence of an enzyme comprising:

a. a solid support comprising a test area surface;

b. a multiplicity of peptide chains each having a proximal end and a distal end, wherein all of said proximal ends of said peptide chains are attached said test area surface of said substrate.

25. The article as recited in claim 24, wherein said solid support is a test strip.

26. The article as recited in claim 25, wherein said test strip comprises a distal end and a proximal end, and wherein said test area surface is disposed proximate to said distal end.

27. The article as recited in claim 24, wherein said solid support is a tube.

28. The article as recited in claim 27, wherein said tube is a glass tube.

29. The article as recited in claim 27, wherein said tube is an Eppendorf tube.

30. The article as recited in claim 24, wherein the majority of said peptide chains each comprise a first tag.

31. The article as recited in claim 30, wherein said first tag is an aromatic alcohol.

32. The article as recited in claim 24, wherein each of said peptide chains comprise an aromatic amino acid.

33. The article as recited in claim 24, wherein said enzyme is pepsin.

34. The article as recited in claim 33, wherein said peptide chains comprise an amino acid residue joined by a peptide bond cleavable by enzymatic hydrolysis.

35. The article as recited in claim 34 wherein said amino acid residue is selected from the group consisting of phenylalanine, tyrosine, tryptophan, and histidine.

36. An apparatus for detecting the presence of an enzyme in a sample of material, said apparatus comprising:

a. an article for exposure to said sample of material, said article comprising a substrate having a test area surface and a multiplicity of peptide chains each having a proximal end and a distal end, wherein all of said proximal ends of said peptide chains are attached said test area surface of said substrate; and

b. a container comprising a fillable volume for receiving a test solution, wherein at least the test area of said article is insertable in said fillable volume.

37. The apparatus as recited in claim 36, wherein said said substrate and said container are unitary.

38. The apparatus as recited in claim 37, wherein said container is a tube.

39. The apparatus as recited in claim 38, wherein said tube is a glass tube.

40. The apparatus as recited in claim 38, wherein said tube is an Eppendorf tube.

41. The apparatus as recited in claim 36, wherein said substrate is a test strip.

42. The apparatus as recited in claim 41, wherein said test strip comprises a distal end and a proximal end, and wherein said test area surface is disposed proximate to said distal end.

43. The apparatus as recited in claim 41, wherein said test strip comprises cellulose fibers.

44. The apparatus as recited in claim 36, wherein the majority of said peptide chains comprise a first tag.

45. The apparatus as recited in claim 44, wherein said first tag is an aromatic alcohol.

46. The apparatus as recited in claim 44, further comprising a test solution.

47. The apparatus as recited in claim 46, wherein said test solution comprises a second tag.

48. The apparatus as recited in claim 47, wherein said second tag is a chemical dye.

49. The apparatus as recited in claim 46, wherein said test solution comprises a buffer.

50. The apparatus as recited in claim 46, wherein said test solution comprises an azo compound.

51. The apparatus as recited in claim 46, wherein said test solution comprises an aromatic diazonium salt.

52. The apparatus as recited in claim 36, further comprising a test solution disposed in said fillable volume of said container.

53. The apparatus as recited in claim 36, further comprising means for obtaining said sample of material.

54. The apparatus as recited in claim 53, wherein said means for obtaining said sample of material comprises a catheter.

55. A process for detecting the presence of an enzyme in a sample of material, said process comprising the steps of:

a. providing an article for exposure to said sample of material, said article comprising a solid support having a test area surface and a multiplicity of peptide chains each having a proximal end and a distal end, wherein all of said proximal ends of said peptide chains are attached to said test area surface of said solid support and said peptide chains are cleavable by an enzyme;

b. exposing said sample of material to said test area surface of said solid support; and

c. measuring an optical property of said test area of said surface of said solid support.

56. The process as recited in claim 55, further comprising the step of first obtaining said sample of material.

57. The process as recited in claim 55, wherein said enzyme is pepsin.

58. A process for detecting the presence of an enzyme in a sample of material, said process comprising the steps of

a. providing an article for exposure to said sample of material, said article comprising a test strip having a test area surface and a multiplicity of peptide chains each having a proximal end and a distal end, wherein all of said proximal ends of said peptide chains are attached said test area surface of said test strip, wherein said peptide chains are cleavable by an enzyme;

b. exposing said sample of material to said test area surface of said test strip; and

c. exposing said test strip to a test solution; and

d. measuring an optical property of said test solution.

59. The process as recited in claim 58, further comprising the step of first obtaining said sample of material.

60. The process as recited in claim 59, wherein said enzyme is pepsin.