Monocolonal antibodies and antigen for human non-small cell lung carcinomas

• US 4,906,562 A
• Filed: 10/18/1985
• Issued: 03/06/1990
• Est. Priority Date: 12/21/1984
• Status: Expired due to Term

• Current Assignee: DADE Behring Beteiligungs GmbH (Siemens AG)
• Original Assignee: Oncogen
• Inventors: Linsley, Peter, Hellstrom, Ingegerd, Brown, Joseph P., Horn, Diane, Hellstrom, Karl E.
• Primary Examiner(s): Moskowitz, Margaret
• Assistant Examiner(s): Kushan, Jeff
• Application Number:
  US06/776,321
• Time in Patent Office: 1,600 Days
• Field of Search: 530/387, 530/389, 530/808, 530/828, 435/7, 435/240.26, 435/240.27, 436/548, 436/514, 436/536, 436/537, 436/542, 436/547
• US Class Current: 435/7.23
• CPC Class Codes: C07K 16/3023 Lung
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: Y10S 530/806 Antigenic peptides or proteins
• CPC Class Codes: Y10S 530/808 Materials and products related to genetic engineering or hybrid or fused cell technology, e.g. hybridoma, monoclonal products
• CPC Class Codes: Y10S 530/828 Cancer
• CPC Class Codes: Y10S 530/866 involving immunoglobulin or antibody fragment, e.g. fab', fab, fv, fc, heavy chain or light chain

Abstract

The present invention is concerned with novel monoclonal antibodies which define a glycolipid antigen associated with human non-small cell lung carcinomas ("NSCLC") and certain other human carcinomas. The antibodies bind to normal human cells to a much lesser degree than to tumor cells. The antibodies find use in diagnostic methods such as the detection of malignant cells associated with NSCLC and in therapeutic methods. Also disclosed in a novel glycolipid antigen. The invention also comprises a method for determining the presence of a malignant condition in lung tissue and other human tissue. The method involves examining the human tissue for the presence of a glycolipid antigen having the terminal carbohydrate sequence: GalNAcβl→4Galβl→3GalNAcβl→4Galβl.fwdarw.R.

Claims

1. A method for determining the presence of a malignant condition in a tissue specimen obtained from an individual, comprising:

(a) contacting the tissue specimen with a monoclonal antibody, the antigen combining site of which competitively inhibits the immunospecific binding of monoclonal antibody L6, produced by hybridoma HB8677 as deposited with the ATCC, to its target antigen; and

(b) detecting whether the monoclonal antibody immunospecifically binds to the tissue specimen, in which immunospecific binding of the monoclonal antibody to the tissue specimen indicates the presence of a malignant condition in the tissue specimen.

2. The method according to claim 1 in which the binding of the monoclonal antibody to the tissue specimen is detected in step (b) by contacting the tissue specimen with a conjugate of a label and a specific binding partner for the monoclonal antibody, the label being capable of producing a detectable signal, so that detection of the signal bound to the tissue specimen indicates the binding of the monoclonal antibody to the tissue specimen.

3. The method according to claim 2 in which the binding partner comprises an antibody specific for the monoclonal antibody.

4. The method according to claim 2 or 3 in which the label comprises a fluorescer, a radiolabel, a chromophore or an enzyme.

5. The method according to claim 1 in which the monoclonal antibody is conjugated to a label capable of producing a detectable signal so that detection of the signal bound to the tissue specimen indicates the binding of the monoclonal antibody conjugate to the tissue specimen.

6. The method according to claim 5 in which the label comprises a fluorescer, a radiolabel, a chromophore or an enzyme.

7. The method according to claim 1 in which the tissue specimen comprises a lung, colon or breast tissue specimen.

8. A method for determining the presence of a maligant condition in a tissue specimen obtained from an individual comprising:

(a) contacting the tissue specimen with an Fab, F(ab'"'"')₂ or Fv fragment of a monoclonal antibody, the antigen combining site of which competitively inhibits the immunospecific binding of monoclonal antibody L6, produced by hybridoma HB8677 as deposited with the ATCC, to its target antigen; and

(b) detecting whether the Fab, F(ab'"'"')₂ or Fv fragment immunospecifically binds to the tissue specimen, in which immunospecific binding of the Fab, F(ab'"'"')₂ or Fv fragment to the tissue specimen indicates the presence of a malignant condition.

9. The method according to claim 8 in which the binding of the Fab, F(ab'"'"')₂ or Fv fragment to the tissue specimen is detected in step (b) by contacting the tissue specimen with a conjugate of a label and a specific binding partner for the Fab, F(ab'"'"')₂ or Fv fragment, the label being capable of producing a detectable signal, so that detection of the signal bound to the tissue specimen indicates the binding of the Fab, F(ab'"'"')₂ or Fv fragment.

10. The method according to claim 9 in which the binding partner comprises an antibody specific for the Fab, F(ab'"'"')₂ or Fv fragment.

11. The method according to claim 9 or 10 in which the label comprises a fluorescer, a radiolabel, a chromophore or an enzyme.

12. The method according to claim 8 in which the Fab, F(ab'"'"')₂ or Fv fragment is conjugated to a label capable of producing a detectable signal so that the detection of the signal bound to the tissue specimen indicates the binding of the Fab, F(ab'"'"')₂ or Fv conjugate to the tissue specimen.

13. The method according to claim 12 in which the label comprises a fluorescer, a radiolabel, a chromophore, or an enzyme.

14. The method according to claim 8 in which the tissue specimen comprises a lung, colon, or breast tissue specimen.

15. A method for determining the presence of a malignant condition in a tissue specimen obtained from an individual comprising:

(a) contacting the tissue specimen with monoclonal antibody L6 which is produced by a hybridoma cell line having the identifying characteristics of HB8677, as deposited with the ATCC; and

(b) detecting whether the monoclonal antibody L6 immunospecifically binds to the tissue specimen, in which binding of the monoclonal antibody L6 to the tissue specimen indicates the presence of a malignant condition in the tissue specimen.

16. The method according to claim 15 in which the binding of monoclonal antibody L6 to the tissue specimen is detected in step (b) by contacting the tissue specimen with a conjugate of a label and a specific binding partner for the monoclonal antibody L6, the label being capable of producing a detectable signal, so that detection of the signal bound to the tissue specimen indicates the binding of the monoclonal antibody L6 to the tissue specimen.

17. The method according to claim 16 in which the binding partner comprises an antibody specific for the monoclonal antibody L6.

18. The method according to claim 16 or 17 in which the label comprises a fluorescer, a radiolabel, a chromophore or an enzyme.

19. The method according to claim 15 in which the monoclonal antibody L6 is conjugated to a label capable of producing a detectable signal so that detection of the signal bound to the tissue specimen indicates the binding of the monoclonal antibody L6 conjugate to the tissue specimen.

20. The method according to claim 19 in which the label comprises a fluorescer, a radiolabel, a chromophore or an enzyme.

21. The method according to claim 15 in which the tissue specimen comprises a lung, colon or breast tissue specimen.

22. A method for determining the presence of a malignant condition in a tissue specimen obtained from an individual, comprising:

(a) contacting the tissue specimen with an Fab, F(ab'"'"')₂ or Fv fragment of monoclonal antibody L6 which is produced by a hybridoma cell line having the identifying characteristics of HB8677, as deposited with the ATCC; and

(b) detecting whether the Fab, F(ab'"'"')₂ or Fv fragment immunospecifically binds to the tissue specimen, in which immunospecific binding of the Fab, F(ab'"'"')₂ or Fv fragment to the tissue specimen indicates the presence of a malignant condition.

23. The method according to claim 22 in which the binding of the Fab, F(ab'"'"')₂ or Fv fragment to the tissue specimen is detected in step (b) by contacting the tissue specimen with a conjugate of a label and a specific binding partner for the Fab, F(ab'"'"')₂ or Fv fragment, the label being capable of producing a detectable signal so that detection of the signal bound to the tissue specimen indicates the binding of the Fab, F(ab'"'"')₂ or Fv fragment to the tissue specimen.

24. The method according to claim 22 in which the binding partner comprises an antibody specific for the Fab, F(ab'"'"')₂ or Fv fragment.

25. The method according to claim 23 or 24 in which the label comprises a fluorescer, a radiolabel, a chromophore or an enzyme.

26. The method according to claim 22 in which the Fab, F(ab'"'"')₂ or Fv fragment is conjugated to a label capable of producing a detectable signal so that the detection of the signal bound to the tissue specimen indicates the binding of the Fab, F(ab'"'"')₂ or Fv conjugate to the tissue specimen.

27. The method according to claim 26 in which the label comprises a fluorescer, a radiolabel, a chromophore, or an enzyme.

28. The method according to claim 22 in which the tissue specimen comprises a lung, colon, or breast tissue specimen.