Recombinant antibodies specific for a growth factor receptor

• US 5,571,894 A
• Filed: 04/29/1994
• Issued: 11/05/1996
• Est. Priority Date: 02/05/1991
• Status: Expired due to Fees

• Current Assignee: Novartis Corporation (Novartis Ag)
• Original Assignee: Ciba-Geigy Corporation (Novartis Ag)
• Inventors: Wels, Winfried S., Harweth, Ina-Maria, Groner, Bernd, Hardman, Norman, Hynes, Nancy E., Zwickl, Markus
• Primary Examiner(s): Walsh, Stephen G.
• Assistant Examiner(s): ULM, JOHN D
• Application Number:
  US08/235,838
• Time in Patent Office: 921 Days
• Field of Search: 435/69.7, 435/252.3, 425/320.1, 530/350, 530/387.3, 536/23.4
• US Class Current: 530/387.3
• 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: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

The invention concerns recombinant antibodies directed to the extracellular domain of the human growth factor receptor c-erbB-2 comprising a light chain variable domain and a heavy chain variable domain of a monoclonal antibody, monoclonal antibodies directed to c-erbB-2 themselves, a method of manufacturing those recombinant and monoclonal antibodies, hybridoma cells secreting those monoclonal antibodies, a method of manufacturing those hybridoma cells, DNAs encoding the heavy and light chain variable domains and the recombinant antibody, a method of manufacturing that DNA, hybrid vectors suitable for the expression of that DNA, host cells transformed with that DNA, and processes of using those recombinant and monoclonal antibodies in the diagnosis and treatment of tumors.

Claims

1. A recombinant antibody directed to the extracellular domain of the growth factor receptor c-erbB-2 comprising of heavy chain variable domain and a light chain variable domain of a monoclonal antibody, wherein the heavy chain variable domain comprises a polypeptide of the formula

View Dependent Claims (2, 3)

2. A recombinant antibody according to claim 1 wherein the heavy chain variable domain comprises a polypeptide of the amino acid sequence 2 to 120 of SEQ ID NO:

5, wherein optionally one or more single amino acids within the amino acid sequences 2 to 31 (FR₁), 37 to 50 (FR₂), 68 to 99 (FR₃), and/or 110 to 120 (FR₄) are replaced by other amino acids or deleted, and wherein the amino acid Cys may be in the oxidized state forming S--S-bridges.

3. A recombinant antibody according to claim 1 wherein the heavy chain variable domain comprises a polypeptide of the amino acid sequence 2 to 120 of SEQ ID NO:

5, wherein the amino acid Cys may be in the oxidized state forming S--S-bridges.

4. A recombinant antibody directed to the extracellular domain of the growth factor receptor c-erbB-2 comprising of heavy chain variable domain and a light chain variable domain of a monoclonal antibody, wherein the light chain variable domain comprises a polypeptide of the formula

View Dependent Claims (5, 6)

5. A recombinant antibody according to claim 4 wherein the light chain variable domain comprises a polypeptide of the amino acid sequence 136 to 241 of SEQ ID NO:

5, wherein optionally one or more single amino acids within the amino acid sequences 136 to 158 (FR₆), 170 to 184 (FR₇), 192 to 223 (FR₈), and/or 233 to 241 (FR₉) are replaced by other amino acids or deleted, and wherein the amino acid Cys may be in the oxidized state forming S--S-bridges.

6. A recombinant antibody according to claim 4 wherein the light chain variable domain comprises a polypeptide of the amino acid sequence 136 to 241 of SEQ ID NO:

5, wherein the amino acid Cys may be in the oxidized state forming S--S-bridges.

7. A recombinant antibody directed to the extracellular domain of the growth factor receptor c-erbB-2 comprising of heavy chain variable domain and a light chain variable domain of a monoclonal antibody, wherein the heavy chain variable domain comprises a polypeptide of the formula

View Dependent Claims (8, 9)

8. A recombinant antibody according to claim 7 wherein the heavy chain variable domain comprises a polypeptide of the amino acid sequence 2 to 121 of SEQ ID NO:

11, wherein optionally one or more single amino acids within the amino acid sequences 2 to 31 (FR₁), 37 to 50 (FR₂), 68 to 99 (FR₃), and/or 111 to 121 (FR₄) are replaced by other amino acids or deleted, and wherein the amino acid Cys may be in the oxidized state forming S--S-bridges.

9. A recombinant antibody according to claim 7 wherein the heavy chain variable domain comprises a polypeptide of the amino acid sequence 2 to 121 of SEQ ID NO:

11, wherein the amino acid Cys may be in the oxidized state forming S--S-bridges.

10. A recombinant antibody directed to the extracellular domain of the growth factor receptor c-erbB-2 comprising of heavy chain variable domain and a light chain variable domain of a monoclonal antibody, wherein the light chain variable domain comprises a polypeptide of the formula

View Dependent Claims (11, 12)

11. A recombinant antibody according to claim 10 wherein the light chain variable domain comprises a polypeptide of the amino acid sequence 137 to 241 of SEQ ID NO:

11, wherein optionally one or more single amino acids within the amino acid sequences 137 to 159 (FR₆), 171 to 185 (FR₇), 193 to 224 (FR₈), and/or 233 to 241 (FR₉) are replaced by other amino acids or deleted, and wherein the amino acid Cys may be in the oxidized state forming S--S-bridges.

12. A recombinant antibody according to claim 10 wherein the light chain variable domain comprises a polypeptide of the amino acid sequence 137 to 241 of SEQ ID NO:

11, wherein the amino acid Cys may be in the oxidized state forming S--S-bridges.

13. A single-chain recombinant antibody directed to the extracellular domain of the growth factor receptor c-erbB-2 comprising of heavy chain variable domain and a light chain variable domain of a monoclonal antibody, wherein the heavy chain variable domain and the light chain variable domain are linked by a polypeptide spacer group, and further wherein the heavy chain variable domain and the light chain variable domain are derived from the mouse monoclonal antibody FRP5.

14. A single-chain recombinant antibody according to claim 13 comprising the heavy chain variable domain of the mouse monoclonal antibody FRP5, the 15 amino acid polypeptide consisting of three repetitive subunits of Gly-Gly-Gly-Gly-Ser, the light chain variable domain of the mouse monoclonal antibody FRP5, and an enzyme or a toxin, or a biologically active variant thereof.

15. A single-chain recombinant antibody designated Fv(FRP5)-phoA according to claim 14 comprising a polypeptide of the amino acid sequence 2 to 690 of SEQ ID NO:

7.

16. A single-chain recombinant antibody designated Fv(FRP5)-ETA according to claim 14 comprising a polypeptide of the amino acid sequence 2 to 606 of SEQ ID NO:

14.

17. A single-chain recombinant antibody directed to the extracellular domain of the growth factor receptor c-erbB-2 comprising of heavy chain variable domain and a light chain variable domain of a monoclonal antibody, wherein the heavy chain variable domain and the light chain variable domain are linked by a polypeptide spacer group, and further wherein the heavy chain variable domain and the light chain variable domain are derived from the mouse monoclonal antibody FWP51.

18. A single-chain recombinant antibody according to claim 17 comprising the heavy chain variable domain of the mouse monoclonal antibody FWP51, the 15 amino acid polypeptide consisting of three repetitive subunits of Gly-Gly-Gly-Gly-Ser, the light chain variable domain of the mouse monoclonal antibody FWP51, and an enzyme or a toxin, or a biologically active variant thereof.

19. A single-chain recombinant antibody designated Fv(FWP51)-ETA according to claim 18 comprising a polypeptide of the amino acid sequence 2 to 606 of SEQ ID NO:

16.