Claims for Patent: 7,585,943

Summary for Patent: 7,585,943

Title:	Compositions and methods for fusion protein separation
Abstract:	The present invention relates to compositions and methods for fusion protein separation utilizing a peptide linker comprising a novel thrombin cleavage site.
Inventor(s):	Kim; Sujeong (Seoul, KR), Kim; Jong-Mook (Seoul, KR), Xu; Song Shan (Beijing, CN)
Assignee:	ViroMed Co., Ltd. (Seoul, KR)
Application Number:	11/407,336
Patent Claims:	1. A chimeric protein comprising a protein of interest, a fusion partner, and a peptide linker interposed there between, wherein said protein of interest is linked to the N-terminus of said peptide linker and said fusion partner is linked to the C-terminus of said peptide linker or said protein of interest is linked to the C-terminus of said peptide linker and said fusion partner is linked to the N-terminus of said peptide linker, wherein the peptide linker consists of the sequence Pro-Arg-Gly-Ser-Pro-Arg-Ala-Ser (SEQ ID NO:7) or the sequence Leu-Val-Pro-Arg-Gly-Ser-Pro-Arg-Ala-Ser (SEQ ID NO:8), and wherein said fusion partner is an affinity peptide. 2. The chimeric protein of claim 1, wherein the peptide linker consists of the sequence Pro-Arg-Gly-Ser-Pro-Arg-Ala-Ser (SEQ ID NO:7). 3. The chimeric protein of claim 1, wherein the peptide linker consists of the sequence Leu-Val-Pro-Arg-Gly-Ser-Pro-Arg-Ala-Ser (SEQ ID NO:8). 4. A chimeric protein comprising a protein of interest, a fusion partner, and a peptide linker interposed there between, wherein said protein of interest is linked to the N-terminus of said peptide linker and said fusion partner is linked to the C-terminus of said peptide linker or said protein of interest is linked to the C-terminus of said peptide linker and said fusion partner is linked to the N-terminus of said peptide linker; wherein the peptide linker consists of the sequence Pro-Arg-Gly-Ser-Pro-Arg-Ala-Ser(SEQ ID NO:7) or the sequence Leu-Val-Pro-Arg-Gly-Ser-Pro-Arg-Ala-Ser (SEQ ID NO:8) wherein said protein of interest is selected from the group consisting of human interleukin (IL)-11, thymosin .beta.4, thymosin .alpha.1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL10, IL-13, IL-15, IL-18, Protease-activated receptor 1 (PAR1), PAR3, PAR4, RANTES, stromal cell-derived factor-1.alpha., monocyte chemotactic protein, stem cell factor, FLT-3L, parathyroid hormone, thrombopoictin, epidermal growth factor, basic fibroblast growth factor, insulin-like growth factor, granulocyte-macrophage colony stimulating factor, granulocyte colony stimulating factor, macrophage colony stimulating factor, platelet-derived growth factor, transforming growth factor (TGF)-.beta.1, tumor necrosis factor (TNF)-.alpha., interferon (IFN)-.alpha., IFN-f.beta., IFN-.gamma., hepatocyte growth factor, vascular endothelial growth factor and immunoglobulin heavy chain, wherein said chimeric protein is non-naturally occurring. 5. The chimeric protein of claim 4, wherein said protein of interest is selected from the group consisting of human IL11, thymosin .beta.4, IL-6 and PAR4. 6. The chimeric protein of claim 4, wherein said protein of interest is human IL11. 7. The chimeric protein of claim 4, wherein said fusion partner is an affinity peptide. 8. The chimeric protein of claim 7, wherein said affinity peptide is selected from the group consisting of glutathione-S-transferase (GST), maltose binding protein (MBP), hexahistidine, T7 peptide, ubiquitin, Flag peptide, c-myc peptide, polyarginine, polycysteine, polyphenylalanine, BTag, galactose binding domain, cellulose binding domain (CBD), thioredoxin, staphylococcal protein A, streptococcal protein G, calmodulin, beta-galactosidase, chloramphenicol acetyltransferase, S-peptide, streptavidin, His-tag, and Strep-tag. 9. The chimeric protein of claim 8, wherein said affinity peptide is selected from the group consisting of GST, MBP and His-tag. 10. The chimeric protein of claim 5, wherein said protein of interest is thymosin .beta.4. 11. The chimeric protein of claim 5, wherein said protein of interest is IL-6. 12. The chimeric protein of claim 5, wherein said protein of interest is PAR4. 13. The chimeric protein of claim 4, wherein the peptide linker consists of the sequence Pro-Arg-Gly-Ser-Pro-Arg-Ala-Ser (SEQ ID NO:7). 14. The chimeric protein of claim 4, wherein the peptide linker consists of the sequence Leu-Val-Pro-Arg-Gly-Ser-Pro-Arg-Ala-Ser (SEQ ID NO:8). 15. The chimeric protein of claim 4, wherein said protein of interest is linked to the N-terminus of said peptide linker and said fusion partner is linked to the C-terminus of said peptide linker. 16. The chimeric protein of claim 4, wherein said protein of interest is linked to the C-terminus of said peptide linker and said fusion partner is linked to the N-terminus of said peptide linker. 17. The chimeric protein of claim 9, wherein said affinity peptide is GST. 18. A chimeric protein comprising a protein of interest, a fusion partner, and a peptide linker interposed there between, wherein said protein of interest is linked to the N-terminus of said peptide linker and said fusion partner is linked to the C-terminus of said peptide linker or said protein of interest is linked to the C-terminus of said peptide linker and said fusion partner is linked to the N-terminus of said peptide linker; said peptide linker consisting of the sequence: Leu-Val-Pro-Arg-Gly-Ser-Pro-Arg-Ala-Ser (SEQ ID NO: 8). 19. The chimeric protein of claim 18, wherein said protein of interest is linked to the N-terminus of said peptide linker and said fusion partner is linked to the C-terminus of said peptide linker. 20. The chimeric protein of claim 18, wherein said protein of interest is linked to the C-terminus of said peptide linker and said fusion partner is linked to the N-terminus of said peptide linker. 21. The chimeric protein of claim 18, wherein said fusion partner is an affinity peptide selected from the group consisting of glutathione-S-transferase (GST), maltose binding protein (MBP), hexahistidine, T7 peptide, ubiquitin, Flag peptide, c-myc peptide, polyarginine, polycysteine, polyphenylalanine, BTag, galactose binding domain, cellulose binding domain (CBD), thioredoxin, staphylococcal protein A, streptococcal protein G, calmodulin, beta-galactosidase, chioramphenicol acetyltransferase, S-peptide, streptavidin, His-tag, and Strep-tag. 22. The chimeric protein of claim 21, wherein said affinity peptide is selected from the group consisting of GST, MBP, and His-tag. 23. The chimeric protein of claim 22, wherein said affinity peptide is GST. 24. The chimeric protein of claim 18, wherein said protein of interest is selected from the group consisting of human IL11, thymosin .beta.4, IL-6 and PAR4. 25. The chimeric protein of claim 1, wherein said affinity peptide is selected from the group consisting of glutathione-S-transferase (GST), maltose binding protein (MBP), hexahistidine, T7 peptide, ubiquitin, Flag peptide, c-myc peptide, polyarginine, polycysteine, polyphenylalanine, BTag, galactose binding domain, cellulose binding domain (CBD), thioredoxin, staphylococcal protein A, streptococcal protein G, calmodulin, beta-galactosidase, chloramphenicol acetyltransferase, S-peptide, streptavidin, His-tag, and Strep-tag. 26. The chimeric protein of claim 25, wherein said affinity peptide is selected from the group consisting of GST, MBP, and His-tag. 27. The chimeric protein of claim 26, wherein said affinity peptide is GST. 28. The chimeric protein of claim 1, wherein said protein of interest is selected from the group consisting of interleukin (IL)-11, thymosin .beta.4, thymosin .alpha.1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-13, IL-15, IL-18, Protease-activated receptor 1 (PAR1), PAR3, PAR4, RANTES, stromal cell-derived factor-1.alpha., monocyte chemotactic protein, stem cell factor, FLT-3L, parathyroid hormone, thrombopoietin, epidermal growth factor, basic fibroblast growth factor, insulin-like growth factor, granulocyte-macrophage colony stimulating factor, granulocyte colony stimulating factor, macrophage colony stimulating factor, platelet-derived growth factor, transforming growth factor (TGF)-.beta.1, tumor necrosis factor (TNF)-.alpha., interferon (IFN)-.alpha., IFN-.beta., IFN-.gamma., hepatocyte growth factor, vascular endothelial growth factor and immunoglobulin heavy chain. 29. The chimeric protein of claim 28, wherein said protein of interest is selected from the group consisting of human IL11, thymosin .beta.4, IL-6 and PAR4. 30. The chimeric protein of claim 1, wherein said protein of interest is linked to the N-terminus of said peptide linker and said fusion partner is linked to the C-tenninus of said peptide linker. 31. The chimeric protein of claim 1, wherein said protein of interest is linked to the C-terminus of said peptide linker and said fusion partner is linked to the N-terminus of said peptide linker.

Details for Patent 7,585,943

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Nps Pharmaceuticals, Inc.	NATPARA	parathyroid hormone	For Injection	125511	01/23/2015		2039-03-29