Transgenically produced fusion proteins

• US 20060026695A1
• Filed: 06/24/2005
• Published: 02/02/2006
• Est. Priority Date: 09/18/1998
• Status: Abandoned Application

• Current Assignee: Dan Pollock, Harry M. Meade, Michael D. Edge, Susanna M. Rybak, Yann Echelard
• Original Assignee: Dan Pollock, Harry M. Meade, Michael D. Edge, Susanna M. Rybak, Yann Echelard
• Inventors: Echelard, Yann, Meade, Harry M., Rybak, Susanna M., Pollock, Dan, Edge, Michael D.
• Application Number:
  US11/166,994
• Publication Number:
  US 20060026695A1
• Time in Patent Office: Days
• US Class Current: 800/4
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/00 Genetically modified animals
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/206 Animal model comprising tissue-specific expression system, e.g. tissue specific expression of transgene, of Cre recombinase
• CPC Class Codes: A01K 2227/10 Mammal
• CPC Class Codes: A01K 2227/101 Bovine
• CPC Class Codes: A01K 2227/102 Caprine
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 2267/0331 Animal model for proliferative diseases
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: C07K 16/04 from milk
• CPC Class Codes: C07K 16/2881 against CD71
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/3007 Carcino-embryonic Antigens
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/54 F(ab')2
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C07K 2319/55 : containing a fusion with a toxin, e.g. diphteria toxin
• CPC Class Codes: C07K 2319/75 : containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones
• CPC Class Codes: C12N 15/62 : DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 15/8509 : for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2830/008 : cell type or tissue specific enhancer/promoter combination

Abstract

A method of making a transgenic fusion protein. The method inlcudes providing a transgenic animal which includes a transgene which provides for the expression of the fusion protein; allowing the transgene to be expressed; and, recovering the fusion protein, from the milk of the transgenic animal.

Claims

1-14. -14. (canceled)

15. An isolated nulceic acid construct, which includes:

a) optionally, an insulator sequence;

b) a mammary epithelial specific promoter;

19. The method of claim 15, wherein the first member of the fusion protein is directly fused to the second member.

20. The method of claim 15, wherein the first member of the fusion protein is linked to the second member by a linker sequence.

21. The method of claim 15, wherein the transgenic mammal is a goat.

22. The method of claim 15, wherein the transgenic mammal is a cow.

16-17. -17. (canceled)

18. A method of making a transgenic fusion protein which includes a first member and a second member, wherein the second member is an enzyme, the method comprising:

providing a non-human transgenic mammal which includes a transgene which provides for the expression of a biologically active fusion protein in the milk of the mammal; and

allowing the transgene to be expressed, thereby providing the fusion protein in the milk of the mammal wherein the second member is in active form and the fusion protein is produced at levels of at least about 0.1 mg/mi in the milk of the mammal wherein the fusion protein includes carboxypeptidase B enzyme and wherein the fusion protein is under the control of a milk-specific protein promoter and has an integrated signal sequence related to that of the milk-specific protein promoter.

23. The transgenic mammal of claim 18, wherein the first member of the fusion protein is an immunoglobulin subunit.

24. The transgenic mammal of claim 18, wherein the first member is fused to the second member and the first member includes the subunit of a targeting molecule and the second member encodes a cell toxin.

25. The transgenic mammal of claim 18, wherein the first member of the fusion protein includes a subunit of an immunoglobulin specific for a tumor antigen.

27. The transgenic mammal of claim 18, wherein the second member of the fusion protein is an RNase.

29. The transgenic mammal of claim 18, wherein the second member of the fusion protein is angiogenin.

30. The transgenic mammal of claim 24, wherein the immunoglobulin subunit of the fusion protein is a human antibody or antigen binding portion thereof.

31. The transgenic mammal of claim 24, wherein the immunoglobulin subunit of the fusion protein is a humanized antibody or antigen binding portion thereof.

32. The transgenic mammal of claim 24, wherein the immunoglobulin subunit of the fusion protein is a chimeric antibody or antigen binding portion thereof.

33. The transgenic mammal of claim 18, wherein the mammal is a goat.

26. The transgenic mammal of claim 26, wherein the tumor antigen is from the group consisting of carcinoembryonic antigen (CEA), a transferrin receptor, TAG-72, and an epidermal growth factor.

35. The method of claim 26, wherein the tumor antigen is a transferrin receptor.

28. The transgenic mammal of claim 28, wherein the RNase is RNase A.