Biomarkers for oxidative stress

• US 6,953,666 B1
• Filed: 11/05/1999
• Issued: 10/11/2005
• Est. Priority Date: 11/06/1998
• Status: Expired due to Fees

• Current Assignee: Emory University
• Original Assignee: Emory University
• Inventors: Le, Ngoc-Anh, Kinkade, Joseph M. Jr., Shapira, Raymond, Jensen, Peter E., Pohl, Jan, Brown, W. Virgil
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): Cook, Lisa V.
• Application Number:
  US09/831,123
• Time in Patent Office: 2,167 Days
• Field of Search: 435/7.1, 435/25, 435/63, 435/69.3, 435/326, 436/71, 436/74, 436/512, 436/547, 514/2, 514/261, 514/474, 514/550, 562/503, 424/85.1, 424/130.1, 530/309, 530/350, 530/351, 530/387.11, 530/387.2, 530/389.1
• US Class Current: 435/7.1
• CPC Class Codes: C07K 16/26 against hormones ; against hormone releasing or inhibiting factors
• CPC Class Codes: G01N 33/6815 containing sulfur, e.g. cysteine, cystine, methionine, homocysteine
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

This invention related generally to methods of detecting and quantifying biomarkers of oxidative stress in proteins. The biomarker may be any amino acid that has undergone oxidation (or other modification, e.g. chloro-tyrosine, dityrosine). Emphasis is given herein on oxidized sulfur- or selenium-containing amino acids (SSAA). The biomarker of oxidative stress in proteins may be detected with an antibody that binds to oxidized amino acids, specifically oxidized sulfur- or selenium-containing amino acids. The antibody may be monoclonal or polyclonal. The presence of biomarker or amount of biomarker present in a sample may be used to aid in assessing the efficacy of environmental, nutritional and therapeutic interventions, among other uses.

Claims

1. An immunoassay method to detect a biomarker of oxidative stress in a biological sample utilizing an antibody which binds said biomarker of oxidative stress, said method comprising the steps of:

(a) contacting a sample containing said biomarker of oxidative stress with said antibody under conditions which allow binding of said biomarker of oxidative stress to said antibody;

(b) detecting the presence of said biomarker of oxidative stress in said sample.

2. The method of claim 1, wherein said biological sample comprises proteins, peptides or proteineaceous aggregates.

3. The method of claim 1, wherein said antibody is bound to a solid phase support.

4. The method of claim 1 further comprising the step of:

(c) comparing the amount of said biomarker of oxidative stress in said sample to a control value for said biomarker of oxidative stress.

5. The method of claim 1 wherein said biomarker of oxidative stress is an oxidized sulfur- or selenium-containing amino acid.

6. The method of claim 5, wherein said oxidized sulfur- or selenium-containing amino acid is selected from the group consisting of the oxidation products of cysteine, cystine, methionine, selenocysteine, selenomethionine and selenocystine.

7. The method of claim 5, wherein said oxidized sulfur- or selenium-containing amino acid is in a sample which comprises proteins, peptides or proteineaceous aggregates.

8. The method of claim 1 wherein said antibody thereof is a polyclonal antibody.

9. The method of claim 8 wherein said polyclonal antibody detects biomarkers of oxidative stress in any protein containing said biomarkers.

10. The method of claim 8 wherein said polyclonal antibody is specific for a protein, peptide or proteineaceous aggregate which includes an oxidized sulfur- or selenium-containing amino acid.

11. The method of claim 8, wherein said polyclonal antibody is from a mouse.

12. The method of claim 1 wherein said antibody is a monoclonal antibody.

15. The method of claim 1, wherein said detection step comprises performing an ELISA.

16. The method of claim 1 wherein picomole levels of biomarker are detected.

17. The method of claim 1, wherein said sample is from an organism selected from the group consisting of:

plants, bacteria, animals, viruses and fungi.

18. The method of claim 17, wherein said sample is mammalian.

19. The method of claim 17, wherein said sample is human.

13. The method of clam 12 wherein said monoclonal antibody detects biomarkers of oxidative stress in any protein containing said biomarkers.

14. The method of claim 13 wherein said monoclonal antibody is specific for a protein, peptide or proteineaceous aggregate which includes an oxidized sulfur- or selenium-containing amino acid.

20. A method of detecting an oxidized sulfur- or selenium-containing amino acid in a biological sample utilizing an antibody which binds a protein, peptide or proteineaceous aggregate which comprises an oxidized sulfur- or selenium-containing amino acid, said method comprising the steps of:

(a) contacting said sample with said antibody under conditions which allow binding of the protein, peptide or proteineaceous aggregate and said antibody; and

(b) detecting the presence of said antibody in said sample.

21. The method of claim 20, wherein said biological sample comprises proteins, peptides or proteineaceous aggregates.

22. The method of claim 20, wherein said antibody is bound to a solid phase support.

23. The method of claim 20 further comprising the step of:

(c) comparing the amount of oxidized sulfur- or selenium-containing amino acid in the sample to a control value for the oxidized sulfur- or selenium-containing amino acid.

24. The method of claim 20, wherein said oxidized sulfur- or selenium-containing amino acid is selected from the group consisting of the oxidation products of cysteine, cystine, methionine, selenocysteine, selenomethionine and selenocystine.

25. The method of claim 20, wherein said oxidized sulfur- or selenium-containing amino acid optionally includes aggregates with other biological material comprising protein, peptides or proteineaceous aggregates.

26. The method of claim 20, wherein said antibody is a monoclonal antibody.

27. The method of claim 26, wherein said monoclonal antibody detects biomarkers of oxidative stress in any protein containing said biomarkers.

28. The method of claim 26, wherein said monoclonal antibody is specific for an oxidized sulfur- or selenium-containing amino acid.

29. The method of claim 20, wherein said antibody is a polyclonal antibody.

30. The method of claim 29, wherein said polyclonal antibody detects biomarkers of oxidative stress in any protein containing said biomarkers.

31. The method of claim 29, wherein said polyclonal antibody is specific for an oxidized sulfur- or selenium-containing amino acid.

32. A method for detecting the presence of oxidative stress in an organism, said method comprising detecting the presence of an antibody that binds an analyte comprising an oxidized sulfur- or selenium containing amino acid, whereby the presence of said antibody is indicative of the presence of oxidative stress in said organism.

33. The method of claim 32, wherein said organism is selected from the group consisting of:

plants, bacteria, animals, viruses and fungi.

34. The method of claim 32, wherein said organism is a mammal.

35. The method of claim 32, wherein said organism is a human.