Rapid Isolation of Osteoinductive Protein Mixtures From Mammalian Bone Tissue

• US 20100041611A1
• Filed: 10/20/2009
• Published: 02/18/2010
• Est. Priority Date: 06/26/2002
• Status: Active Grant

• Current Assignee: Kevin Thorne, Rama Akella
• Original Assignee: Kevin Thorne, Rama Akella
• Inventors: AKELLA, Rama, Thorne, Kevin
• Granted Patent: US 8,829,166 B2
• Time in Patent Office: Days
• US Class Current: 514/1.100
• CPC Class Codes: A61K 35/32 Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
• CPC Class Codes: A61K 38/1875 Bone morphogenic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: C07K 1/34 by filtration, ultrafiltration or reverse osmosis
• CPC Class Codes: C07K 1/36 by a combination of two or more processes of different types
• CPC Class Codes: C07K 14/51 Bone morphogenetic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
• CPC Class Codes: Y10S 530/84 Bones; tendons; teeth; cartilage

Abstract

A method for purifying bone-derived osteoinductive proteins including a demineralization process, a protein extraction process, a high molecular weight ultrafiltration process, a low molecular weight ultrafiltration process, and a recover process. The high and low ultrafiltration processes preferably select proteins having a nominal molecular weight between approximately 8 kilodaltons and approximately 100 kilodaltons. Processes of the present invention may be used to recover osteoinductive proteins from bone demineralization waste streams.

Claims

1-16. -16. (canceled)

17. A method of producing an osteoinductive device, comprising:

contacting bone tissue with an acidic demineralization medium to provide demineralized bone tissue and a mineralized supernatant;

separating the mineralized supernatant from the demineralized bone tissue;

recovering osteoinductive proteins from the mineralized supernatant; and

combining the osteoinductive proteins with a carrier.

18. The method of claim 17, wherein the carrier comprises a solid.

19. The method of claim 18, wherein the solid carrier is selected from the group consisting of collagen, a bone derivative, hydroxyapatite or a composite device.

20. The method of claim 18, wherein the solid carrier comprises a bone derivative selected from the group consisting of bone chips or bone matrix.

21. The method of claim 18, wherein the solid carrier comprises bone matrix.

22. The method of claim 21, wherein the bone matrix comprises demineralized and devitalized bone matrix (DVBM).

23. The method of claim 17, wherein the carrier comprises a liquid.

24. The method of claim 23, wherein the liquid carrier is selected from the group consisting of saline, a blood-derivative or bone marrow aspirate.

25. The method of claim 23, wherein the liquid carrier comprises a blood derivative selected from the group consisting of whole blood, serum, plasma or platelet rich plasma (PRP).

26. The method of claim 18, wherein the liquid carrier comprises serum.

27. The method of claim 18, wherein the liquid carrier comprises saline.

28. The method of claim 17, wherein the carrier comprises a slurry.

29. The method of claim 17, further comprising storing the osteoinductive proteins as a solution or a solid.

30. The method of claim 29, wherein the osteoinductive proteins are stored sterile.

31. The method of claim 29, wherein the osteoinductive proteins are stored under vacuum or an inert gas atmosphere.

32. The method of claim 31, wherein the inert gas is selected from the group consisting of nitrogen, hydrogen, helium, argon or mixtures thereof.

33. The method of claim 29, wherein the osteoinductive proteins are stored as a solid.

34. The method of claim 33, wherein the osteoinductive proteins are lyophilized.

35. The method of claim 17, further comprising removing at least some mineral from the mineralized supernatant.

36. The method of claim 35, wherein removing comprises performing tangential flow filtration of the mineralized supernatant.

37. The method of claim 35, wherein removing comprises contacting the mineralized supernatant with mineral precipitation agent.