Glycosylation in avians

• US 20090178147A1
• Filed: 01/07/2009
• Published: 07/09/2009
• Est. Priority Date: 01/07/2008
• Status: Active Grant

• Current Assignee: Alexion Pharmaceuticals Incorporated (Astrazeneca PLC)
• Original Assignee: Synageva BioPharma Corp (Astrazeneca PLC)
• Inventors: Harvey, Alex J.
• Granted Patent: US 8,431,770 B2
• Time in Patent Office: Days
• US Class Current: 800/4
• CPC Class Codes: A01K 2217/052 inducing gain of function
• CPC Class Codes: A01K 2227/30 Bird
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2799/027 where the vector is derived from a retrovirus
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination
• CPC Class Codes: C12N 9/1048 Glycosyltransferases (2.4)
• CPC Class Codes: C12N 9/1051 Hexosyltransferases (2.4.1)

Abstract

Transgenic avians which produce proteins in their oviduct tissue having modified oligosaccharide structures and methods of making such avians are disclosed herein. The invention also includes the modified proteins produced in the transgenic birds.

Claims

1. A transgenic avian containing a transgene comprising a glycosyltransferase coding sequence wherein oviduct tissue of the transgenic avian produces protein having an N-linked oligosaccharide comprising at least one saccharide wherein the oligosaccharide would not comprise the saccharide in the absence of the transgene.

2. The transgenic avian of claim 1 wherein the glycosyltransferase is a N-acetylglucosaminyltransferase.

3. The transgenic avian of claim 1 wherein the glycosyltransferase is a N-acetylglucosaminyltransferase 3.

4. The transgenic avian of claim 1 wherein the oviduct tissue is magnum tissue.

5. The transgenic avian of claim 1 wherein the oviduct tissue comprises tubular gland cells.

6. The transgenic avian of claim 1 wherein cells of the oviduct tissue secrete the protein in the presence of egg white.

7. The transgenic avian of claim 1 wherein the transgene comprises an LTR.

8. The transgenic avian of claim 1 wherein the transgene comprises an oviduct specific promoter.

9. A method comprising isolating the protein produced by the transgenic avian of claim 1.

10. The transgenic avian of claim 1 wherein the protein is exogenous to the avian.

11. A method comprising isolating the exogenous protein produced by the transgenic avian of claim 10.

12. The transgenic avian of claim 1 wherein the protein is a therapeutic protein.

13. The method of claim 1 wherein the avian is a chicken.

14. A transgenic avian containing a transgene comprising a galactosyltransferase coding sequence wherein oviduct tissue of the transgenic avian produces protein having an N-linked oligosaccharide comprising at least one galactose wherein the oligosaccharide would not comprise the galactose in the absence of the transgene.

15. The transgenic avian of claim 14 wherein the glycosyltransferase is galactosyltransferase type 1.

16. The transgenic avian of claim 14 wherein the protein is produced in tubular gland cells.

17. The transgenic avian of claim 14 wherein cells of the oviduct tissue secrete the protein in the presence of egg white.

18. The transgenic avian of claim 14 wherein the oligosaccharide comprises 1 to 5 galactoses.

19. The transgenic avian of claim 14 wherein the transgene comprises an oviduct specific promoter.

20. A method comprising isolating the protein produced by the transgenic avian of claim 14.

21. The transgenic avian of claim 14 wherein the protein is an exogenous protein.

22. A method comprising isolating the exogenous protein produced by the transgenic avian of claim 21.

23. The method of claim 22 wherein the protein is a therapeutic protein.

24. The method of claim 22 wherein the avian is a chicken.

25. A transgenic avian containing a transgene comprising a sialyltransferase coding sequence wherein oviduct tissue of the transgenic avian produces protein having an N-linked oligosaccharide comprising at least one sialic acid wherein the oligosaccharide would not comprise the sialic acid in the absence of the transgene.

26. The transgenic avian of claim 25 wherein the glycosyltransferase is galactosyltransferase type 1.

27. The transgenic avian of claim 25 wherein the protein is produced in tubular gland cells.

28. The transgenic avian of claim 25 wherein cells of the oviduct tissue secrete the protein in the presence of egg white.

29. The transgenic avian of claim 25 wherein the oligosaccharide comprises 1 to 5 sialic acids.

30. The transgenic avian of claim 25 wherein the transgene comprises an oviduct specific promoter.

31. A method comprising isolating the protein produced by the transgenic avian of claim 25.

32. The transgenic avian of claim 25 wherein the protein is an exogenous protein.

33. A method comprising isolating the exogenous protein produced by the transgenic avian of claim 32.

34. The method of claim 33 wherein the protein is a therapeutic protein.

35. The method of claim 33 wherein the avian is a chicken.

36. A method of producing a protein in an avian wherein the protein is exogenous to the avian, the method comprising producing a transgenic avian containing a transgene encoding a glycosyltransferase wherein oviduct tissue of the avian produces an exogenous protein encoded by a second transgene and having an N-linked oligosaccharide comprising at least one of a galactose and a sialic acid wherein the oligosaccharide would not comprise the galactose and/or sialic acid in the absence of the transgene encoding a glycosyltransferase.