Apparatus for the detection of pepsin

• US 7,402,423 B2
• Filed: 08/13/2004
• Issued: 07/22/2008
• Est. Priority Date: 08/13/2004
• Status: Expired due to Fees

• Current Assignee: Biomed Solutions, LLC
• Original Assignee: Biomed Solutions, LLC
• Inventors: Bucholtz, Kevin, Horner, Scott, Taghizadeh, Farhan
• Primary Examiner(s): GITOMER, RALPH J
• Application Number:
  US10/918,562
• Publication Number:
  US 20060035307A1
• Time in Patent Office: 1,439 Days
• Field of Search: 435/24
• US Class Current: 435/287.1
• 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 article for detecting the presence of pepsin suitable to test a subject in a physician'"'"'s office, the subject suspected of suffering from gastroesophageal reflux disease and providing a sample suspected of containing pepsin, the article comprising:

a. a solid support comprising a test area surface;

b. a multiplicity of peptide chains each having a peptide proximal end and a peptide distal end,

wherein the peptide distal end comprises a peptide bearing a first tag that interacts with a second tag to provide a detectable chromophoric product,

wherein all of said peptide proximal ends are attached to said test area surface of said support, and

wherein said peptide chains are cleavable by said pepsin to provide said peptide proximal end attached to said test area surface and said peptide distal end bearing the detectable first tag cleaved from said test area surface.

2. The article as recited in claim 1, wherein said solid support is a test strip.

3. The article as recited in claim 2, 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.

4. The article as recited in claim 1, wherein said solid support is a tube.

5. The article as recited in claim 4, wherein said tube is a glass tube.

6. The article as recited in claim 4, wherein said tube is an Eppendorf tube.

7. The article as recited in claim 1, wherein said detectable first tag comprises an aromatic alcohol.

8. The article as recited in claim 1, wherein each of said peptide chains comprises an aromatic amino acid.

9. The article as recited in claim 1, wherein said peptide chains comprise an amino acid residue joined by a peptide bond cleavable by said pepsin.

10. The article as recited in claim 9 wherein said amino acid residue is selected from the group consisting of phenylalanine, tyrosine, tryptophan, and histidine.

11. The article described in claim 10 wherein said amino acid residue is preceded by an asparagine residue and followed by an asparagine residue.

12. The article described in claim 1 wherein the peptide chain comprises the sequence —

Y—

X—

Y—

, wherein each Y is independently any amino acid residue other than valine, alanine, or glycine, and X is an aromatic amino acid.

13. The article described in claim 12 wherein X is selected from the group consisting of phenylalanine, tyrosine, tryptophan, and histidine.

14. The article described in claim 12 wherein each Y is asparagine.

15. The article described in claim 1 wherein the article detects the presence of pepsin in about 2 hours or less.

16. The article described in claim 1 wherein the second tag comprises an aromatic diazonium salt.

17. The article described in claim 1 wherein the chromophoric product is detectable by the human eye, thereby permitting visual detection of pepsin.

18. The article described in claim 1 further comprising means for obtaining said sample of material.

19. The article described in claim 18 wherein said means for obtaining said sample of material comprises a catheter.

20. An apparatus for detecting the presence of pepsin in a sample of material suitable to test a subject in a physician'"'"'s office, the subject suspected of suffering from gastroesophageal reflux disease and providing the sample, 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 peptide proximal end and a peptide distal end,

wherein the peptide distal end comprises a peptide bearing a detectable first tag,

wherein all of said peptide proximal ends are attached to said test area surface of said substrate, and

wherein said peptide chains are cleavable by said pepsin to provide said peptide proximal end attached to said test area surface and said peptide distal end bearing the detectable first tag cleaved from said test area surface; and

b. a container comprising a fillable volume for receiving a test solution comprising at least one of the sample, a buffer and a second tag that interacts with the first tag to provide a detectable chromophoric product, and wherein at least the test area of said article is insertable in said fillable volume.

21. The apparatus as recited in claim 20, wherein said substrate and said container are unitary.

22. The apparatus as recited in claim 21, wherein said container is a tube.

23. The apparatus as recited in claim 22, wherein said tube is a glass tube or an Eppendorf tube.

24. The apparatus as recited in claim 20, wherein said substrate is a test strip.

25. The apparatus as recited in claim 24, wherein said test strip comprises a strip distal end and a strip proximal end, and wherein said test area surface is disposed proximate to said distal end.

26. The apparatus as recited in claim 24, wherein said test strip comprises cellulose fibers.

27. The apparatus as recited in claim 20, wherein said detectable first tag comprises an aromatic alcohol.

28. The apparatus as recited in claim 20, further comprising the test solution disposed in the fillable volume.

29. The apparatus as recited in claim 28, wherein said test solution comprises the second tag.

30. The apparatus as recited in claim 29, wherein said second tag is a chemical dye.

31. The apparatus as recited in claim 28, wherein said test solution comprises a buffer.

32. The apparatus as recited in claim 28, wherein said test solution comprises an aromatic diazonium salt.

33. The apparatus as recited in claim 20, further comprising a test solution disposed in said fillable volume of said container.

34. The apparatus as recited in claim 20, further comprising means for obtaining said sample of material.

35. The apparatus as recited in claim 34, wherein said means for obtaining said sample of material comprises a catheter.

36. The apparatus described in claim 20 wherein the peptide chain comprises the sequence —

Y—

X—

Y—

, wherein each Y is independently any amino acid residue other than valine, alanine, or glycine, and X is an aromatic amino acid.

37. The apparatus described in claim 36 wherein X is selected from the group consisting of phenylalanine, tyrosine, tryptophan, and histidine.

38. The apparatus described in claim 36 wherein each Y is asparagine.

39. The apparatus described in claim 20 wherein the apparatus detects the presence of pepsin in a sample of material in about 2 hours or less.

40. The apparatus described in claim 20 wherein the chromophoric product is detectable by the human eye, thereby permitting visual detection of pepsin.

41. The apparatus described in claim 20 wherein each of said peptide chains comprises an aromatic amino acid.

42. The apparatus described in claim 20 wherein said peptide chains comprise an amino acid residue joined by a peptide bond cleavable by said pepsin.

43. The apparatus described in claim 42 wherein said amino acid residue is selected from the group consisting of phenylalanine, tyrosine, tryptophan, and histidine.

44. The apparatus described in claim 43 wherein said amino acid residue is preceded by an asparagine residue and followed by an asparagine residue.