Anti-human tenascin monoclonal antibody

• US 20110020219A1
• Filed: 09/17/2008
• Published: 01/27/2011
• Est. Priority Date: 02/26/2002
• Status: Active Grant

• Current Assignee: Alfasigma S.p.A.
• Original Assignee: Sigma-Tau Industrie Farmaceutiche Riunite SPA (Alfasigma S.p.A.)
• Inventors: Anna Maria, Anastasi, Rita, De Santis
• Granted Patent: US 8,048,417 B2
• Time in Patent Office: Days
• US Class Current: 424/1.49
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: G01N 2333/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
• CPC Class Codes: G01N 33/574 for cancer
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/975 Kit

Abstract

A novel anti-human tenascin ST2146 monoclonal antibody is described endowed with high affinity with the native antigen and high tumor selectivity. The cST2146 hybridoma is stably producing the antibody in high density culture conditions and is suitable for the industrial development of ST2146-based products. ST2146 exhibits properties exploitable for both therapeutic and diagnostic applications.

Claims

1. A monoclonal antibody to human tenascin C, whose light chain variable region comprises at least one amino acid sequence selected from the group consisting SEQ ID NO:

9, SEQ ID NO;

11 and SEQ ID NO;

13 and heavy chain variable region comprises at least one amino acid sequence selected from the group consisting of SEQ ID NO;

15, SEQ ID NO;

17 and SEQ ID NO;

19, or a fragment of said monoclonal antibody, wherein said antibody or fragment binds to an antigenic epitope within the EGF-like repeat of human tenascin C.

2. A conjugate comprising the monoclonal antibody or the fragment of claim 1, said conjugate further comprising at least one marker, diagnostic agent or biotin.

3. The antibody or fragment of claim 1, wherein said antibody or fragment is produced recombinantly.

4. The antibody or fragment of claim 1, which comprises a human constant region.

5. The conjugate of claim 2, wherein said conjugate further comprises a member of the avidin family or a cytokine.

6. The conjugate of claim 2, wherein said conjugate further comprises a toxin, a superantigen or a cytokine.

9. The conjugate of claim 2, wherein said conjugate further comprises an antitumor drug or a radioisotope.

12. The hybridoma cell line producing the antibody of claim 1, deposited with Advanced Biotechnology Center on 29 Jan. 2002 under the provision of Budapest Treaty, with the accession number PD02003.

13. A process for the preparation of antibody, comprising culturing the hybridoma cell line of claim 12 and isolating said antibody.

22. A therapeutic kit comprised of five vials, whose first vial contains a conjugate of claim 2, the second vial contains an avidin, the third vial contains biotinylated albumin, the fourth vial contains a streptavidin, and the fifth vial contains a radiolabelled biotin or biotin derivative.

23. The therapeutic kit according to claim 22, wherein the contents of said vials are suitable for human injection.

28. A method of determining levels of circulating tenascin comprising contacting an antibody or fragment of claim 1, said antibody or fragment produced recombinantly, with a second tenascin-specific antibody in a sandwich assay in the presence of a patient sample and determining the level of circulating tenascin.

29. A diagnostic kit comprising the antibody or fragment of claim 1.

30. A container comprising the conjugate of claim 2, and a buffer or a reagent.

31. The container according to claim 30, further comprising a separate tenascin-specific antibody which binds the A-D fragment.

32. The container according to claim 30, further comprising a separate tumor specific antibody.

33. The antibody of claim 1 in combination with a second tenascin-specific antibody, where said second antibody binds to a second antigenic epitope of tenascin.

34. Pharmaceutical composition comprising the antibody or fragment of claim 1 in admixture with at least one pharmaceutically acceptable excipient or vehicle.

35. The conjugate of claim 2, wherein said antibody or fragment is produced recombinantly.

36. The conjugate of claim 2 which comprises a human constant region.

37. The antibody or fragment of claim 1, wherein said fragment is proteolytic.

38. The container according to claim 30, wherein said container contains multiple compartments.

39. A method of determining levels of circulating tenascin comprising contacting a conjugate of claim 2 with a second tenascin-specific antibody in a sandwich assay in the presence of a patient sample and determining the level of circulating tenascin.

40. The container according to claim 31, wherein said tenascin-specific antibody which binds the A-D fragment is in a separate compartment of said container.

41. The container according to claim 32, wherein said tumor specific antibody is in a separate compartment of said container.

42. A DNA encoding the antibody of claim 1, or a fragment thereof.

43. A DNA encoding the light chain variable region of the antibody of claim 1.

44. A DNA encoding the heavy chain variable region of the antibody of claim 1.

7-8. -8. (canceled)

10-11. -11. (canceled)

14-21. -21. (canceled)

24-27. -27. (canceled)