Claims for Patent: 6,984,772

Summary for Patent: 6,984,772

Title:	Transgenic non-human mammals producing fibrinogen in their milk
Abstract:	A transgenic, non-human mammalian animal is capable of expressing a heterologous gene for human or other recombinant physiologically functional fibrinogen holoprotein or individual subunit chain polypeptides thereof or a modified or fusion fibrinogen in mammary glands of the animals and secreting the expressed product into a body fluid. Methodology employing such a mammal yields recombinant physiologically functional fibrinogens, subunit chain polypeptides thereof, and modified or fusion fibrinogens.
Inventor(s):	Velander; William H. (Blacksburg, VA), Drohan; William N. (Springfield, VA), Lubon; Henryk (Rockville, MD), Johnson; John L. (Blacksburg, VA)
Assignee:	Virginia Tech Intellectual Properties, Inc. (Blacksburg, VA) American Red Cross (Rockville, MD) The University of North Carolina at Chapel Hill (Chapel Hill, NC)
Application Number:	08/443,184
Patent Claims:	1. A method for producing biologically active fibrinogen comprising: providing a transgenic non-human mammal whose genome comprises a first DNA segment encoding a first DNA encoding a heterologous fibrinogen A.alpha. chain, a second DNA segment encoding a second DNA encoding a heterologous fibrinogen B.beta. chain, and a third DNA segment encoding a third DNA encoding a heterologous fibrinogen .gamma. chain, wherein each chain is derived from the same species, and wherein each of said first, second and third segments is operably linked to a cis-acting, expression promoter-containing regulatory sequence required for its expression in a mammary gland of a female transgenic non-human mammal; allowing the expression of said first, second and third DNA segments and the production of milk containing biologically active fibrinogen in said female mammal; collecting milk from said female mammal; and recovering the biologically active fibrinogen from the milk. 2. The method according to claim 1, wherein said transgenic non-human mammal is selected from the group consisting of a rodent, rabbit, sheep, pig, goat and cattle. 3. The method according to claim 2, wherein said transgenic non-human mammal is a sheep. 4. The method according to claim 1, wherein said transgenic non-human mammal is a cow. 5. The method according to claim 1, wherein said promoter to which each of said first, second and third DNA segments is operably linked is selected from the group consisting of casein, .beta.-lactoglobulin, .alpha.-lactalbumin and whey acidic protein gene promoters. 6. The method according to claim 5, wherein said promoter is a .beta.-lactoglobulin promoter. 7. The method according to claim 5, wherein said promoter is a casein promoter. 8. The method according to claim 5, wherein said promoter is a whey acidic protein gene promoter. 9. The method according to claim 1, wherein said fibrinogen is human fibrinogen. 10. The method according to claim 1, wherein said first, second, and third DNA segments comprises an intron. 11. A method for producing biologically active fibrinogen comprising: providing a first DNA segment encoding a heterologous fibrinogen .alpha.a chain, a second DNA segment encoding a heterologous fibrinogen B.beta. chain; and a third DNA segment encoding a heterologous fibrinogen .gamma. chain, wherein each chain is from the same species, and wherein each of said first, second and third segments is operably linked to a promoter-containing regulatory sequence required for its expression in the mammary gland of a female transgenic non-human mammal; introducing said DNA segments into a fertilized egg of a non-human mammalian species heterologous to the species of origin of said fibrinogen chains; inserting said egg into an oviduct or uterus of a female of said mammalian species to obtain a transgenic non-human mammal whose genome comprises said DNA segments; breeding said mammal to produce female progeny that express said first, second and third DNA segments and produce milk containing biologically active fibrinogen encoded by said segments; collecting milk from said female progeny; and recovering the biologically active fibrinogen from the milk. 12. The method according to claim 11, wherein said transgenic non-human mammal is selected from the group consisting of a rodent, rabbit, sheep, pig, goat and cattle. 13. The method according to claim 11, wherein said species into which said DNA segments is introduced is a sheep. 14. The method according to claim 11, wherein said species into which said DNA segments is introduced is a cow. 15. The method according to claim 11, wherein said non-human mammal is a cow. 16. The method according to claim 11, wherein said first, second, and third DNA segments comprises an intron. 17. The method according to claim 11, wherein each of said first, second and third DNA segments is operably linked to a transcription promoter selected from the group consisting of casein, .beta.-lactoglobulin, .alpha.-lactalbumin and whey acidic protein gene promoters. 18. A method according to claim 11, wherein said first, second and third DNA segments are expressed under the control of a .beta.-lactoglobulin promoter. 19. The method according to claim 11, wherein said first, second and third DNA segments are expressed under the control of a casein promoter. 20. The method according to claim 11, wherein said first, second and third DNA segments are expressed under the control of a whey acidic protein gene promoter. 21. The method according to claim 11, wherein said introducing step comprises injecting said first, second and third DNA segments into a pronucleus of said fertilized egg. 22. The method according to claim 11, wherein said fibrinogen is human fibrinogen. 23. The method according to claim 11, wherein said introducing step comprises injecting said first, second and third DNA segments into a pronucleus of a fertilized egg and inserting said egg into an oviduct of a pseudopregnant female to produce a female transgenic non-human mammal whose genome comprises said DNA segments, wherein said egg and said pseudopregnant female are of the same species. 24. The method of claim 23, wherein said female mammal is a sheep. 25. The method of claim 23, wherein said female mammal is a cow. 26. A transgenic non-human mammal, wherein the genome of said mammal comprises: a first DNA segment encoding a heterologous fibrinogen A.alpha. chain, a second DNA segment encoding a heterologous fibrinogen B.beta. chain, a third DNA segment encoding a heterologous fibrinogen .gamma. chain, and further wherein each chain is derived from the same species and is operably linked to a promoter-containing regulatory sequence required for its expression in the mammary gland of a host female mammal, wherein expression of said DNA segments results in the production of recoverable quantities of biologically active fibrinogen from milk of a female of said non-human mammal. 27. The non-human transgenic mammal according to claim 26, wherein said non-human mammal is selected from the group consisting of a rodent, rabbit, sheep, pig, goat and cattle. 28. The non-human transgenic mammal according to claim 27, wherein said non-human mammal is a sheep. 29. The non-human transgenic mammal according to claim 26, wherein said non-human mammal is a cow. 30. The non-human transgenic mammal according to claim 26, wherein each of said first, second and third DNA segments is operably linked to a transcription promoter selected from the group consisting of casein, .beta.-lactoglobulin, .alpha.-lactalbumin and whey acidic protein gene promoters. 31. The non-human transgenic mammal according to claim 26, wherein said first, second and third DNA segments are expressed under the control of a .beta.-lactoglobulin promoter. 32. The non-human transgenic mammal according to claim 26, wherein said first, second and third DNA segments are expressed under the control of a casein promoter. 33. The non-human transgenic mammal according to claim 26, wherein said first, second and third DNA segments are expressed under the control of a whey acidic protein gene promoter. 34. The non-human mammal produced according to claim 26, wherein said mammal is female. 35. The non-human mammal produced according to claim 26, wherein said mammal is male. 36. A process for producing a transgenic non-human mammal comprising: providing a first DNA segment encoding a first DNA encoding a heterologous fibrinogen A.alpha. chain, a second DNA segment encoding a second DNA encoding a heterologous fibrinogen B.beta. chain, and a third DNA segment encoding a third DNA encoding a heterologous fibrinogen .gamma. chain, wherein each chain is derived from the same species, and wherein each of said first, second and third segments is operably linked to a promoter-containing regulatory sequence required for its expression in a mammary gland of a host female mammal; introducing said DNA segments into a fertilized egg of a non-human species heterologous to the species of origin of said fibrinogen chains; inserting said fertilized egg into an oviduct or uterus of a female of said mammalian species; and allowing said fertilized egg to develop thereby producing a transgenic non-human mammal whose genome comprises said first, second and third DNA segments, wherein female progeny of said mammal express said DNA segments in a mammary gland to produce biologically active fibrinogen in recoverable quantities in the milk of said female progeny. 37. The process according to claim 36, wherein said mammal is female. 38. The process according to claim 36, wherein said mammal is male.

Details for Patent 6,984,772

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2023-01-10
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2023-01-10
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2023-01-10
Csl Behring Gmbh	RIASTAP	fibrinogen concentrate (human)	For Injection	125317	01/16/2009		2023-01-10
Octapharma Pharmazeutika Produktionsges.m.b.h.	FIBRYGA, FIBRYNA	fibrinogen (human)	For Injection	125612	06/07/2017		2023-01-10