Claims for Patent: 7,307,148

Summary for Patent: 7,307,148

Title:	Method for purification of albumin conjugates
Abstract:	The present invention relates to a method for separating albumin conjugate from unconjugated albumin in a solution comprising albumin conjugate and unconjugated albumin by loading the solution onto a hydrophobic support equilibrated in aqueous buffer having a high salt content; applying to the support a gradient of decreasing salt concentration; and collecting the eluted albumin conjugate.
Inventor(s):	Bousquet-Gagnon; Nathalie (St-Jerome, CA), Quraishi; Omar (Hudson, CA), Bridon; Dominique P. (San Francisco, CA)
Assignee:	ConjuChem Biotechnologies Inc. (Montreal (Quebec), CA)
Application Number:	11/112,277
Patent Claims:	1. A method for separating albumin conjugate from unconjugated albumin in a solution comprising albumin conjugate and unconjugated albumin, the method comprising: a) loading said solution onto a hydrophobic interaction chromatography matrix equilibrated in aqueous buffer at a salt concentration high enough to promote matrix-protein interactions; b) applying to said matrix a gradient of decreasing salt concentration; and c) collecting said albumin conjugate. 2. The method of claim 1, wherein said albumin conjugate consists of a molecule comprising a Michael acceptor covalently bonded to albumin. 3. The method of claim 2, wherein said Michael acceptor is a maleimide group. 4. The method of claim 3, wherein said maleimide group is maleimid-propionic acid. 5. The method of claim 1, wherein said albumin is selected from the group consisting of serum albumin and recombinant albumin. 6. The method of claim 1, wherein said albumin is selected from the group consisting of human albumin, rat albumin, mouse albumin, swine albumin, bovine albumin, dog albumin, and rabbit albumin. 7. The method of claim 1, wherein said albumin is human serum albumin. 8. The method of claim 1, wherein said albumin is modified with a fatty acid, a metal ion, a sugar, or a combination thereof. 9. The method of claim 8, wherein said sugar is selected from the group consisting of glucose, lactose, and mannose. 10. The method of claim 2, wherein said molecule is selected from the group consisting of a peptide, a DNA, an RNA, and a combination thereof, to which said Michael acceptor is covalently bonded, optionally through a linker. 11. The method of claim 10, wherein said molecule is a peptide having a molecular weight of at least 57 daltons. 12. The method of claim 10, wherein said peptide is selected from the group consisting of glucagon like peptide 1 (GLP-1), atrial natriuretic peptide (ANP), kringle 5 (KS), dynorphin, exendin-4, growth hormone releasing factor (GRF), insulin, natriuretic peptides, enfuvirtide (T-20), T-1249, C-34, soluble C-35 peptide BK (SC-35), peptide YY (PYY), and analogs thereof. 13. The method of claim 10, wherein said molecule is selected from the group consisting of: 3',4'-didehydro-4'-deoxy-C'-norvincaleukoblastine; 2'-deoxy-2',2'-difluorocytidine; and 5.beta.,20-epoxy-1 ,2.alpha.,4,7.beta.,10.beta.,13.alpha.-hexahydroxy-tax-11-en-9-one, 4,10-diacetate 2-benzoate, 13-ester with (2R,3S)-N-benzoyl-3-phenylisoserine. 14. The method of claim 10, wherein said molecule is covalently attached to said albumin through an acid sensitive covalent bond or a peptide sequence susceptible to proteolytic cleavage, thereby allowing the separation of said molecule and albumin and the entry of the molecule into a cell. 15. The method of claim 1, wherein said hydrophobic interaction chromatography matrix is a column containing a hydrophobic resin. 16. The method of claim 15, wherein said hydrophobic resin is a bead-formed agarose-based gel filtration matrix covalently coupled to a ligand selected from the group consisting of an octyl group, a phenyl group, and a butyl group. 17. The method of claim 15, wherein said hydrophobic resin is a bead-formed agarose-based gel filtration matrix covalently coupled to a butyl group. 18. The method of claim 1, wherein said gradient of decreasing salt concentration has a starting salt concentration of between 500 and 3000 mM. 19. The method of claim 1, wherein said salt is selected from the group consisting of ammonium phosphate, ammonium sulfate, and magnesium phosphate. 20. The method of claim 1, wherein said salt is ammonium phosphate or ammonium sulfate. 21. The method of claim 1, wherein said salt is ammonium sulfate. 22. The method of claim 2, wherein said bond is between said Michael acceptor and cysteine 34 of said albumin. 23. The method of claim 2, 3, or 4, wherein said molecule is a peptide to which said Michael acceptor is covalently bonded, optionally through a linker. 24. The method of claim 23, wherein said peptide is an analog of glucagon like peptide 1 (GLP-1), glucagon like peptide 2 (GLP-2), atrial natriuretic peptide (ANP), kringle 5 (KS), dynorphin, exendin-4, growth hormone releasing factor (GRF), insulin, natriuretic peptides, enfuvirtide (T-20), T-1249, C-34, soluble C-35 peptide EK (SC-35), or peptide YY (PYY). 25. The method of claim 2, 3, or 4, wherein said molecule is selected from the group consisting of GLP-1 (7-36) dAla.sup.8 Lys.sup.37 (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:1),GRF (1-29) dAla.sup.2 Gin.sup.8 Ala.sup.15 Leu.sup.27 Lys.sup.30 (.epsilon.-MPA) CONH.sub.2 (SEQ ID NO:2), Ac-K5 Lys.sup.8 (.epsilon.-MPA)-NH.sub.2 (SEQ ID NO:3), Insulin B1-MPA (SEQ ID NO:4), Insulin A1-MPA (SEQ ID NO:5), MPA-AEEA-C34-CONH.sub.2 (SEQ ID NO:6), C34 (1-34) Lys.sup.35 (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:7), C34 (1-34) Lys.sup.13 (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:8), GLP-1 (7-36) Lys.sup.37 (.epsilon.-MPA)-NH.sub.2 (SEQ ID NO:9), GLP-1 (7-36) dAla.sup.8 Lys.sup.37 (.epsilon.-MPA)-NH.sub.2 (SEQ ID NO:10), GLP-1 (7-36) Lys.sup.26 (.epsilon.-AEEA-AEEA-MPA) (SEQ ID NO:11), GLP-1 (7-36) Lys.sup.34 (.epsilon.-AEEA-AEEA-MPA) (SEQ ID NO:12), Exendin-4-(1-39) Lys.sup.40 (.epsilon.-MPA)-NH.sub.2 (SEQ ID NO:13), Exendin-4 (9-39) Lys.sup.40 (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:14), Dyn A (1-13) (MPA)-NH.sub.2 (SEQ ID NO:15), MPA-AEEA-ANP (99-126)-CONH.sub.2 (SEQ ID NO:16), Dyn A (7-13) Lys.sup.13 (.epsilon.-MPA)-CONH.sub.2 (SEQ ID NO:17), acetyl-Phe-His-cyclohexylstatyl-lle-Lys (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:18), GLP-1 (7-36) Lys.sup.23 (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:19), GLP-1 (7-36) Lys.sup.18 (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:20), GLP-1 (7-36) Lys.sup.26 (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:21), GLP-1 (7-37) Lys.sup.27 (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:22), GLP-1 (7-36) Lys.sup.37 (.epsilon.-AEEA-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:23), GLP-1 (7-36) Lys.sup.37 (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:24), Exendin-4-(1-39) Lys.sup.40 (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:25), GLP-1 (7-36) Lys.sup.34 (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:26), Insulin B1-OA-MPA (SEQ ID NO:27), Insulin B29-MPA (SEQ ID NO:28), GRF (1-29) Lys.sup.30 (.epsilon.-MPA)-CONH.sub.2 (SEQ ID NO:29), GRF (1-29) dAla.sup.2 Gln.sup.8 dArg.sup.11 Ala.sup.15 Leu.sup.27 Lys.sup.30 (.epsilon.-MPA)-CONH.sub.2 (SEQ ID NO:30), GRF (1-29) dAla.sup.2 Lys.sup.30 (.epsilon.-MPA)-CONH.sub.2 (SEQ ID NO:31), GLP-1 (9-36) Lys.sup.37 (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:32), Ac-T20 (1-36) Lys.sup.37 (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:33), Ac-T1249 (1-39) Lys.sup.40 (.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:34), 3',4'-didehydro-4'-deoxy-C'-norvincaleukoblastine -AEEA-MPA, C34 (1-34) Lys.sup.13 (.epsilon.-MPA)-CONH.sub.2 (SEQ ID NO:36), C34 (1-34) Lys.sup.35 (.epsilon.-MPA)-CONH.sub.2 (SEQ ID NO:37), MPA-C34 (1-34)-CONH.sub.2 (SEQ ID NO:38), Ac-C34 (1-34) Glu.sup.2 Lys.sup.6 Lys.sup.7 Glu.sup.9 Glu.sup.10 Lys.sup.13 Lys.sup.14 Glu.sup.16 Glu.sup.17 Lys.sup.20 Lys.sup.21 Glu.sup.23 Glu.sup.24 Lys.sup.27 Glu.sup.31 Lys.sup.34 Lys.sup.35 Lys.sup.36(.epsilon.-AEEA-MPA)-CONH.sub.2 (SEQ ID NO:39), MPA-AEEA-C34 (1-34) Glu.sup.2 Lys.sup.6 Lys.sup.7 Glu.sup.9 Glu.sup.10 Lys.sup.13 Lys.sup.14 Glu.sup.16 Glu.sup.17 Lys.sup.21 Glu.sup.23 Glu.sup.24 Lys.sup.27 Glu.sup.31 Lys.sup.34 Lys.sup.35 CONH.sub.2 (SEQ ID NO:40), PYY (3-36) Lys.sup.4 (.epsilon.-OA-MPA)-CONH.sub.2 (SEQ ID NO:41), MPA-OA-PYY (3-36)-CONH.sub.2 (SEQ ID NO:42), Insulin B29-AEES2-MPA (SEQ ID NO:43), Insulin B1-AEES2-MPA (SEQ ID NO:44), Insulin B29-OA-MPA (SEQ ID NO:45), MPA-PYY (3-36)-CONH.sub.2 (SEQ ID NO:46), PYY (3-36) Lys.sup.37 (.epsilon.-MPA)-CONH.sub.2 (SEQ ID NO:47), MPA-PYY (22-36)-CONH.sub.2 (SEQ ID NO:48), Acetyl-PYY (22-36) Lys.sup.37 (.epsilon.-MPA)-CONH.sub.2 (SEQ ID NO:49), MPA-ANP (99-126)-CONH.sub.2 (SEQ ID NO:50), MPA-EEEEP-ANP (99-126) (SEQ ID NO:51), and GLP-2 (1-33) Gly.sup.2 Lys.sup.34 (.epsilon.-MPA)-CONH.sub.2 (SEQ ID NO:52). 26. The method of claim 24, wherein said peptide is GLP-1 (7-36) dAla.sup.8 Lys.sup.37. 27. The method of claim 24, wherein said peptide is Exendin-4 (1-39) Lys.sup.40. 28. The method of claim 2, 3, or 4, wherein said molecule is GLP-1 (7-36) dAla.sup.8 Lys.sup.37 (.epsilon.-AEEA-MPA)-CONH.sub.2.

Details for Patent 7,307,148

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Grifols Therapeutics Llc	ALBUKED, PLASBUMIN-20, PLASBUMIN-25, PLASBUMIN-5	albumin (human)	For Injection	101138	10/21/1942		2024-04-23
Baxalta Us Inc.	BUMINATE, FLEXBUMIN	albumin (human)	Injection	101452	03/03/1954		2024-04-23
Csl Behring Ag	ALBURX	albumin (human)	Injection	102366	07/23/1976		2024-04-23
Grifols Biologicals Llc	ALBUTEIN	albumin (human)	Injection	102478	08/15/1978		2024-04-23
Instituto Grifols, S.a.	HUMAN ALBUMIN GRIFOLS	albumin (human)	Injection	103352	02/17/1995		2024-04-23
Instituto Grifols, S.a.	HUMAN ALBUMIN GRIFOLS	albumin (human)	Injection	103352	06/11/2003		2024-04-23