Method for the diagnosis and prognosis of malignant diseases

• US 7,541,150 B2
• Filed: 04/08/2002
• Issued: 06/02/2009
• Est. Priority Date: 04/08/2002
• Status: Expired due to Fees

• Current Assignee: University of Louisville
• Original Assignee: University of Louisville
• Inventors: Bates, Paula J., Xu, Xiaohua, Trent, John O., Miller, Donald M.
• Primary Examiner(s): Yaen; Christopher H
• Application Number:
  US10/118,854
• Publication Number:
  US 20030194754A1
• Time in Patent Office: 2,612 Days
• Field of Search: 436/64, 436/500, 435/7.1
• US Class Current: 435/7.1
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: G01N 33/57492 involving compounds localized on the membrane of tumor or cancer cells

Abstract

Methods for the diagnosis of cancer by determining the neoplastic status of a cell by probing the cell plasma membrane for the presence of nucleolin are provided, as are kits to carry out such tests.

Claims

1. A method of determining presence of a malignant cell in a sample directly from a patient, comprising:

obtaining a sample comprising a primary cell of the patient; and

detecting the presence of plasma membrane nucleolin in the primary cell in the sample, to determine presence of the malignant cell.

2. The method of claim 1, wherein the primary cell is mammalian.

3. The method of claim 2, wherein the primary cell is monkey, dog, cat, rabbit, cow, pig, goat, guinea pig, mouse, rat or sheep.

4. The method of claim 2, wherein the primary cell is human.

5. The method of claim 1, wherein the primary cell is lysed.

6. The method of claim 1 wherein the detecting comprises detecting nucleolin-anti-nucleolin agent complex.

7. The method of claim 6, wherein the anti-nucleolin agent is a monoclonal antibody that specifically binds nucleolin.

8. The method of claim 7, wherein the antibody is p7-1A4 mouse monoclonal antibody, sc-8031, sc-9893, sc-9892, 4E2 or 3G4B2.

9. The method of claim 6, wherein the complex is detected by detecting fluorescence, an enzyme, or radioactivity.

18. The method of claim 1, wherein the primary cell is a lung cell.

19. The method of claim 1, wherein the sample comprises sputum.

20. The method of claim 1, further comprising, prior to the detecting:

sending the sample to a testing center.

10. A method of determining presence of a malignant cell in a sample directly from a patient, comprising:

obtaining a sample comprising a primary cell of the patient; and

quantifying the amount of plasma membrane nucleolin in a primary cell of the patient in the sample, to determine presence of the malignant cell.

11. The method of claim 10, wherein the primary cell is mammalian.

12. The method of claim 11, wherein the primary cell is monkey, dog, cat, rabbit, cow, pig, goat, guinea pig, mouse, rat or sheep.

13. The method of claim 12, wherein the primary cell is human.

14. The method of claim 10, wherein the primary cell is lysed.

15. The method of claim 10 wherein the quantifying comprises quantifying nucleolin-anti-nucleolin agent complex.

16. The method of claim 15, wherein the anti-nucleolin agent is a monoclonal antibody.

17. The method of claim 15, wherein the antibody is p7-1A4 mouse monoclonal antibody, sc-8031, sc-9893, sc-9892, 4E2 or 3G4B2.

21. The method of claim 10, wherein the primary cell is a lung cell.

22. The method of claim 10, wherein the sample comprises sputum.

23. The method of claim 10, further comprising, prior to the quantifying:

collecting the sample directly from the patient; and

sending the sample to a testing center.

24. A method of determining presence of a malignant cell in a sample directly from a patient, comprising:

obtaining a sample comprising a primary cell of the patient; and

detecting the presence of plasma membrane nucleolin in the primary cell in the sample, to determine presence of the malignant cell;

wherein the detecting of the presence of plasma membrane nucleolin comprises;

contacting the primary cell with an antibody that specifically binds nucleolin; and

detecting the antibody bound to nucleolin, to detect the presence of plasma membrane nucleolin in the primary cell.

25. The method of claim 24, wherein the detecting of the presence of plasma membrane nucleolin comprises quantifying the amount of plasma membrane nucleolin in the primary cell.

26. The method of claim 25, wherein the primary cell is human.

27. The method of claim 26, wherein the antibody is a monoclonal antibody.

28. The method of claim 27, wherein the antibody is p7-1A4 mouse monoclonal antibody, sc-8031, sc-9893, sc-9892, 4E2 or 3G4B2.

29. The method of claim 26, wherein the detecting of the antibody bound to nucleolin comprises detecting fluorescence.

30. The method of claim 26, wherein the primary cell is a lung cell.

31. The method of claim 26, wherein the sample comprises sputum.

32. The method of claim 26, further comprising, prior to the detecting of the presence of plasma membrane nucleolin:

sending the sample to a testing center.

33. The method of claim 32, wherein the primary cell is a lung cell, and the sample comprises sputum.

34. A method of determining if a patient has cancer, comprising:

obtaining a sample comprising a primary cell of the patient;

detecting the presence of plasma membrane nucleolin in the primary cell in the sample; and

diagnosing the patient as having cancer.