Method for making a monoclonal antibody, monoclonal antibodies to .alpha. P D

• US 5,468,468 A
• Filed: 06/25/1993
• Issued: 11/21/1995
• Est. Priority Date: 02/09/1989
• Status: Expired due to Term

• Current Assignee: United States Department Of Health And Human Services
• Original Assignee: United States Department Of Health And Human Services
• Inventors: Aaronson, Stuart A., LaRochelle, William J., Jensen, Roy A., Pierce, Jacalyn
• Primary Examiner(s): Lacey, David L.
• Assistant Examiner(s): LORING, SUSAN
• Application Number:
  US08/081,216
• Time in Patent Office: 879 Days
• Field of Search: 530/387.2, 530/387.3, 530/388.1, 530/388.22, 530/387.1, 530/391.3, 424/143.1, 424/9, 424/1.49, 435/240.27
• US Class Current: 424/1.49
• CPC Class Codes: A61K 2123/00 Preparations for testing in vivo
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 51/103 against receptors for growth factors or receptors for growth regulators
• CPC Class Codes: C07K 14/71 for growth factors; for growth regulators
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/2863 against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 2317/77 Internalization into the cell
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: G01N 2333/49 Platelet-derived growth factor [PDGF]
• CPC Class Codes: G01N 2333/71 for growth factors; for growth regulators
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds
• CPC Class Codes: G01N 33/74 involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors

Abstract

Potent neutralizing monoclonal antibodies to the human α PDGF receptor (α PDGFR) and fragments thereof are described. These monoclonal antibodies specifically bind to an epitope on α PDGFR, inhibits PDGF binding with PDGF, antagonizes PDGF, and does not bind β PDGFR receptor. A hybridoma cell line producing such a monoclonal antibody, methods of in vivo imaging of a pathological conditions and methods of inhibiting the growth of a neoplasia expressing α PDGFR, which use these monoclonal antibodies are also described. In vitro assays for detecting the presence of α PDGFR and for evaluating the binding affinity of a test compound are also described.

Claims

1. A monoclonal antibody or fragment thereof which specifically binds to an epitope on α

PDGF receptor, inhibits PDGF binding with PDGF receptor, antagonizes PDGF and does not bind β

PDGF receptor.

2. The monoclonal antibody or fragment thereof of claim 1 which has a half-maximal binding affinity of 50 picomolar, as measured by solid phase ELISA.

3. The monoclonal antibody or fragment thereof of claim 1 which is a humanized antibody.

4. The monoclonal antibody or fragment thereof of claim 1 which is a chimeric antibody.

5. The monoclonal antibody or fragment thereof of claim 1 which is a single chain antibody.

6. The monoclonal antibody or fragment thereof of claim 1 which is an anti-idiotype antibody.

7. The monoclonal antibody or fragment thereof of claim 1 which is α

R1 deposited as ATCCHB 11390.

8. The monoclonal antibody or fragment thereof of claim 1 which is labeled for detection.

9. A hybridoma cell line which secretes the monoclonal antibody of claim 1.

10. A pharmaceutical composition for in vivo imaging of a pathological condition that expresses α

PDGF receptors, comprising;

(a) the monoclonal antibody or fragment thereof of claim 8 which binds α

PDGF receptor in vivo; and

(b) a pharmaceutically acceptable carrier.

11. An in vivo method of imaging a pathological condition that express α

PDGF receptors comprising the steps of;

(a) administering to a subject an imaging-effective amount of the labeled monoclonal antibody or fragment thereof of claim 8 and a pharmaceutically effective carrier; and

(b) detecting said the binding of said labeled monoclonal antibody or fragment thereof to said α

PDGF receptors in said pathological condition.

12. The method of claim 11, wherein said pathological condition is selected from the group consisting of neoplasia, arteriosclerosis, arthritis, scleroderma, bone marrow fibrosis, and keloids.

13. The method of claim 12, wherein said monoclonal antibody or fragment thereof is radiolabeled.

14. The method of claim 12, wherein said detecting involves radioactive imaging.

15. A method of producing the monoclonal antibody of claim 1 comprising immunizing Balb/c mice with an interleukin 3 dependent mouse hematopoietic cell transfactant that expresses human α

PDGFR at the cell surface.