Claims for Patent: RE39293

Summary for Patent: RE39293

Title:	Separation of plasma components
Abstract:	A method for separating components from plasma, the method comprising (I) separating the plasma into a first and second component, the first component comprising an albumin/.alpha.-1-antirypsin pool and the second component comprising plasma containing components having a molecular mass greater than albumin; (II) treating the second component to form an immunoglobulins concentrate containing immunoglobulins substantially free from components having a molecular mass less than immunoglobulins; (III) treating the immunoglobulins concentrate to remove components having a molecular mass greater than immunoglobulins; and (IV) separating albumin and .alpha.-1-antitrypsin from the albumin/.alpha.-1-antitrypsin pool.
Inventor(s):	Gilbert; Andrew Mark (Lane Cove, AU), Conlan; Brendon (Binya, AU), Nair; Chenicheri Hariharam (Baulkham Hills, AU)
Assignee:	Life Therapeutics Limited (Frenchs Forest, AU)
Application Number:	10/632,644
Patent Claims:	1. A method of separating components from plasma, the method comprising the steps: (a) placing the plasma in a first solvent stream, the first solvent stream being separated from a second solvent stream by a first electrophoretic separation membrane having a molecular mass cut-off less than the molecular mass of albumin and a restriction membrane having a molecular mass cut-off less than the first electrophoretic separation membrane; (b) selecting a buffer for the firm solvent stream having a pH greater than the pI of albumin; (c) applying an electric potential between the two solvent streams causing movement of albumin and .alpha.-1-antitrypsin through the first electrophoretic membrane into the second solvent stream while biomolecules having a molecular mass greater than albumin and .alpha.-1-antitrypsin are substantially retained in the first solvent stream, or if entering the first electrophoresis membrane, being substantially prevented from passing through the first electrophoresis membrane, wherein biomolecules in the plasma having a molecular mass less than albumin and .alpha.-1-antitrypsin are caused to move through the first separation membrane and the restriction membranes to a waste collection; (d) optionally, periodically stopping and reversing the electric potential to cause movement of biomolecules having a molecular mass greater than albumin and .alpha.-1-antitrypsin having entered the first electrophoresis membrane to move back into the first solvent stream, wherein substantially not causing any albumin or .alpha.-1-antitrypsin that have entered the second solvent stream to re-enter first solvent stream; (e) maintaining steps (c) and optionally (d) until the desired amount of albumin and .alpha.-1-antitrypsin have been collected as an albumin/.alpha.-1-antitrypsin pool and biomolecules having a molecular mass less than albumin and .alpha.-1-antitrypsin have been removed from the first solvent stream to form a treated plasma; (f) placing the treated plasma in a third solvent stream, the third solvent stream being separated from a fourth solvent stream by a second electrophoretic separation membrane having a molecular mass cut-off less than the molecular mass of immunoglobulins; (g) selecting a buffer for the third solvent stream having a pH above neutral; (h) applying an electric potential between the third and fourth solvent streams causing movement of biomolecules having a molecular mass less than that of immunoglobulins in the treated plasma through the second electrophoretic separation membrane into the fourth solvent stream while immunoglobulins and other biomolecules having a molecular mass greater than immunoglobulins are substantially retained in the third solvent stream, or if entering the second electrophoresis separation membrane, being substantially prevented from passing through the second electrophoresis separation membrane; (i) optionally, periodically stopping and reversing the electric potential to cause movement of immunoglobulins and other biomolecules having a molecular mass greater than immunoglobulins having entered the second electrophoresis separation membrane to move back into the third solvent stream, wherein substantially not causing any biomolecules having a molecular mass less than immunoglobulins that have entered the fourth solvent stream to re-enter third solvent stream; (j) maintaining steps (h) and optional (i) until the desired amount of biomolecules having a molecular mass less than immunoglobulins have been removed from the third upstream to form an immunoglobulins concentrate; (k) removing the biomolecules from the fourth solvent stream; (l) replacing the second electrophoretic separation membrane with a third electrophoretic separation membrane having a molecular mass cut-off greater than the molecular mass of immunoglobulins; (m) selecting a buffer for the immunoglobulins concentrate having a pH below neutral; (n) applying an electric potential between the immunoglobulins concentrate in the third solvent stream and a fresh fourth solvent stream causing movement of immunoglobulins in the immunoglobulins concentrate in the third solvent stream through the third electrophoretic separation membrane into the fresh fourth solvent stream while biomolecules having a molecular mass greater than immunoglobulins are substantially retained in the third solvent stream, or if entering the third electrophoresis separation membrane, being substantially prevented from passing through the third electrophoresis separation membrane; (o) optionally, periodically stopping and reversing the electric potential to cause movement of biomolecules having a molecular mass greater than immunoglobulins having entered the third electrophoresis membrane to move back into the treated third solvent stream, wherein substantially not causing any immunoglobulins that has entered the fresh fourth solvent stream to re-enter treated third solvent stream; (p) maintaining steps (n) and optional (o) until the desired amount of immunoglobulins have been moved to the fresh fourth downstream; (q) placing the albumin/.alpha.-1-antitrypsin concentrate in a fifth solvent stream, the fifth solvent stream being separated from a sixth solvent stream by a fourth electrophoretic separation membrane having a molecular mass cut-off less than the molecular mass of albumin; (r) selecting a buffer for the fifth solvent stream having a pH greater than neutral; (s) applying an electric potential between the fifth and sixth solvent streams causing movement of .alpha.-1-antitrypsin through the fourth electrophoresis separation membrane into the sixth solvent stream while albumin is substantially retained in the fifth solvent stream, or if entering the fourth electrophoresis separation membrane, being substantially prevented from passing through the fourth electrophoresis separation membrane; (t) optionally, periodically stopping and reversing the electric potential to cause movement of albumin having entered the fourth electrophoresis separation membrane to move back into the fifth solvent stream, wherein substantially not causing any .alpha.-1-antitrypsin that has entered the sixth solvent stream to re-enter the fifth solvent stream; and (u) maintaining steps (s) and optionally (t) until the desired amount of albumin remains in the fifth solvent stream and the desired amount of .alpha.-1-antitrypsin has have been removed to the sixth solvent stream..]. .[.2. The method according to claim 1 wherein steps (q) to (u) are carried out after steps (a) to (e)..]. .[.3. The method according to claim 1 wherein the plasma is a pooled human plasma sample..]. .[.4. The method according to claim 1 wherein the first electrophoresis separation membrane of step (a) has molecular mass cut-off of about 75 kDa and the restriction membrane has a molecular mass cut off of about 50 kDa..]. .[.5. The method according to claim 1 wherein the buffer in step (b) has a pH of 9..]. .[.6. The method according to claim 5 wherein the buffer is a Tris-borate buffer..]. .[.7. The method according to claim 1 wherein the second electrophoresis separation membrane of step (f) has a molecular mass cut-off of 200 kDa..]. .[.8. The method according to claim 1 wherein the third electrophoresis separation membrane of step (l) has a molecular mass cut-off of 500 kDa..]. .[.9. The method according to claim 1 wherein the buffer of the third solvent stream in step (g) has a pH of 9..]. .[.10. The method according to claim 1 wherein the buffer of the immunoglobulins concentrate of step (m) has a pH of less than 5..]. .[.11. The method according to claim 10 wherein buffer has a of pH 4.6..]. .[.12. The method according to claim 1 wherein the fourth electrophoresis separation membrane of step (q) has molecular mass cut-off of about 50 kDa..]. .[.13. The method according to claim 1 wherein the buffer of the fifth solvent stream in step (r) has a pH of 8.0..]. .[.14. The method according to claim 13 wherein the buffer is a Tris-borate buffer..]. .[.15. The method according to claim 1 wherein a potential of 250 volts is applied in steps (c), (h), (n) and (s)..]. .[.16. The method according to claim 1 wherein the immunoglobulins are immunoglobulin G (IgG)..]. .[.17. The method according to claim 1 wherein yields of albumin, immunoglobulins and .alpha.-1-antitrypsin from plasma are at least 70% and purity of at least 90%..]. .[.18. The method according to claim 1 wherein albumin, immunoglobulins and .alpha.-1-antitrypsin are separated from plasma in less than 1 day..]. .[.19. The method according to claim 18 wherein albumin, immunoglobulins and .alpha.-1-antitrypsin are separated from plasma in less than 12 hours..]. .[.20. The method according to claim 18 wherein albumin, immunoglobulins and .alpha.-1-antitrypsin are separated from plasma in less than 6 hours..]. .Iadd.21. A method for separating plasma components from a plasma sample containing at least albumin, .alpha.-1-antitrypsin, and immunoglobulins, by electrophoresis, comprising: (I) separating the plasma into a first and second component using a first electrophoretic separation membrane whereby the first component containing a mixture of albumin and .alpha.-1-antitrypsin resides on one side of the first electrophoretic separation membrane while the second component resides on the other side of the first electrophoretic separation membrane; (II) separating the second component into third and fourth components whereby the third component containing immunoglobulins is located on one side of a second electrophoretic separation membrane, and the fourth component resides on the other side of the second separation membrane; (III) removing material having a molecular mass greater than immunoglobulins from the third component using a third electrophoretic separation membrane whereby immunoglobulins reside on one side of the third electrophoretic separation membrane and material having a molecular mass greater than immunoglobulins reside on the other side of the third electrophoretic separation membrane; and (IV) separating albumin from .alpha.-1-antitrypsin in the first component using a fourth separation membrane whereby .alpha.-1-antitrypsin resides on one side of the fourth electrophoretic separation membrane and albumin resides on the other side of the fourth separation membrane..Iaddend. .Iadd.22. The method according to claim 21, whereby the second component contains material having a molecular mass greater than albumin..Iaddend. .Iadd.23. The method according to claim 21, whereby the first component migrates through the first electrophoretic separation membrane..Iaddend. .Iadd.24. The method according to claim 21, whereby the fourth component contains material having a molecular mass less than immunoglobulins contained in the third component..Iaddend. .Iadd.25. The method according to claim 21, whereby .alpha.-1-antitrypsin migrates through the fourth electrophoretic separation membrane..Iaddend. .Iadd.26. The method according to claim 21, whereby step (I) further comprises: (a) placing the plasma in a first solvent stream, the first solvent stream being separated from a second solvent stream by the first electrophoretic separation membrane having a molecular mass cut-off more than the molecular mass of albumin, and the second solvent stream being further bounded by a restriction membrane having a molecular mass cut off less than the first electrophoretic separation membrane; (b) selecting a buffer for the first solvent stream having a pH greater than the pI of albumin; (c) applying an electric potential between the two solvent streams whereby albumin and .alpha.-1-antitrypsin migrate through the first electrophoretic membrane into the second solvent stream while material having a molecular mass greater than albumin and .alpha.-1-antitrypsin are substantially prevented from passing through the first electrophoretic membrane; (d) optionally, periodically stopping and reversing the electric potential whereby material having a molecular mass greater than albumin and .alpha.-1-antitrypsin that have entered the first electrophoretic membrane move back into the first solvent stream, while substantially preventing albumin or .alpha.-1-antitrypsin in the second solvent stream from re-entering the first solvent stream; and (e) maintaining steps (c) and optionally (d) until the desired amount of albumin and .alpha.-1-antitrypsin migrates into the second solvent stream..Iaddend. .Iadd.27. The method according to claim 26, whereby the material having a molecular mass less than albumin and .alpha.-1-antitrypsin move through the first separation membrane and the restriction membrane..Iaddend. .Iadd.28. The method according to claim 21, whereby step (II) further comprises: (f) placing the second component in a third solvent stream, the third solvent stream being separated from a fourth solvent stream by a second electrophoretic separation membrane having a molecular mass cut-off less than the molecular mass of immunoglobulins; (g) selecting a buffer for the third solvent stream having a pH above neutral; (h) applying an electric potential between the third and fourth solvent streams whereby immunoglobulins from the second component are substantially prevented from passing through the second electrophoretic separation membrane thereby forming the third component while material having a molecular mass less that that of immunoglobulins in the second component migrate through the second electrophoretic separation membrane into the fourth solvent stream to form the fourth component; (i) optionally, periodically stopping and reversing the electric potential whereby materials from the third component that have entered the second electrophoretic separation membrane move back into the third solvent stream while preventing materials from the fourth component from re-entering the third solvent stream; and (j) maintaining steps (h) and optional (i) until the desired amount of third component has been separated from the fourth component..Iaddend. .Iadd.29. The method according to claim 28 whereby step (III) further comprises: (l) replacing the second electrophoretic separation membrane with a third electrophoretic separation membrane having a molecular mass cut-off greater than the molecular mass of immunoglobulins; (m) selecting a buffer for the third solvent stream having a pH below neutral; (n) replacing the fourth solvent stream with a fresh fourth solvent stream; (o) applying an electric potential between the third solvent stream and the fresh fourth solvent stream whereby immunoglobulins in the third component migrate through the third electrophoretic separation membrane into the fresh fourth solvent stream; (p) optionally, periodically stopping and reversing the electric potential whereby material having a molecular mass greater than immunoglobulins in the third component that have entered the third electrophoretic membrane move back into the third solvent stream while preventing immunoglobulins in the fresh fourth solvent stream from re-entering the third solvent stream; and (q) maintaining steps (o) and optional (p) until the desired amount of immunoglobulins migrate to the fresh fourth solvent stream..Iaddend. .Iadd.30. The method according to claim 21, whereby step (IV) further comprises: (r) placing the first component in a fifth solvent stream, the fifth solvent stream being separated from a sixth solvent stream by a fourth electrophoretic separation membrane having a molecular mass cut-off less than the molecular mass of albumin; (s) selecting a buffer for the fifth solvent stream having a pH greater than neutral; (t) applying an electric potential between the fifth and sixth solvent streams whereby .alpha.-1-antitrypsin migrates through the fourth electrophoretic separation membrane into the sixth solvent stream while albumin is substantially prevented from passing through the fourth electrophoretic separation membrane; (u) optionally, periodically stopping and reversing the electric potential whereby albumin that has entered the fourth electrophoretic separation membrane moves back into the fifth solvent stream while preventing .alpha.-1-antitrypsin in the sixth solvent stream from re-entering the fifth solvent stream; and (v) maintaining steps (t) and optionally (u) until the desired amounts of albumin and .alpha.-1-antitrypsin are separated on opposite sides of the fourth separation membrane..Iaddend. .Iadd.31. The method according to claim 21, further comprising using more than one electrophoretic separation apparatus..Iaddend. .Iadd.32. The method according to claim 21, further comprising using three electrophoretic separation apparatus..Iaddend. .Iadd.33. The method according to claim 6 whereby the first electrophoretic separation membrane of step (a) has molecular mass cut-off of about 75 kDa and the restriction membrane has a molecular mass cut off of about 50 kDa..Iaddend. .Iadd.34. The method according to claim 26 whereby the first electrophoretic separation membrane of step (a) has a molecular mass cut-off greater than 67 kDa..Iaddend. .Iadd.35. The method according to claim 26 whereby the buffer in step (b) has a pH of about 9..Iaddend. .Iadd.36. The method according to claim 26 whereby the buffer is a Tris-borate buffer..Iaddend. .Iadd.37. The method according to claim 26 whereby the electric potential applied in step (c) is 250 volts..Iaddend. .Iadd.38. The method according to claim 28 whereby the second electrophoretic separation membrane of step (f) has a molecular mass cut-off of about 200 kDa..Iaddend. .Iadd.39. The method according to claim 28 whereby the second electrophoretic separation membrane of step (f) has a molecular mass cut-off greater than 150 kDa..Iaddend. .Iadd.40. The method according claim 28 whereby the buffer of the third solvent stream in step (g) has a pH of about 9..Iaddend. .Iadd.41. The method according to claim 28 where the electric potential applied in step (h) is 250 volts..Iaddend. .Iadd.42. The method according to claim 29 whereby the third electrophoretic separation membrane of step (l) has a molecular mass cut-off of about 500 kDa..Iaddend. .Iadd.43. The method according to claim 29 whereby the buffer of the immunoglobulins concentrate of step (m) has a pH of less than 5..Iaddend. .Iadd.44. The method according to claim 29 whereby buffer has a pH of about 4.6..Iaddend. .Iadd.45. The method according to claim 29 whereby the electric potential applied in step (o) is 250 volts..Iaddend. .Iadd.46. The method according to claim 30 whereby the fourth electrophoretic separation membrane of step (q) has molecular mass cut-off of about 50 kDa..Iaddend. .Iadd.47. The method according to claim 30 whereby the fourth electrophoretic separation membrane of step (q) has molecular mass cut-off less than 54 kDa..Iaddend. .Iadd.48. The method according to claim 30 whereby the buffer of the fifth solvent stream in step (r) has a pH of about 8.0..Iaddend. .Iadd.49. The method according to claim 30 whereby the buffer is a Tris-borate buffer..Iaddend. .Iadd.50. The method according to claim 30 whereby the electric potential applied in step (t) is 250 volts..Iaddend. .Iadd.51. The method according to any one of claims 21-50, whereby albumin, immunoglobulin, and .alpha.-1-antitrypsin are separated from plasma..Iaddend. .Iadd.52. The method according to any one of claims 21-50 whereby the immunoglobulins are immunoglobulin G (IgG)..Iaddend. .Iadd.53. The method according to any one of claims 21-50 whereby yields of albumin, immunoglobulins and .alpha.-1-antitrypsin from plasma are at least 70% and purity of at least 90%..Iaddend. .Iadd.54. The method according to any one of claims 21-50 whereby albumin, immunoglobulins and .alpha.-1-antitrypsin are separated from plasma in less than 1 day..Iaddend. .Iadd.55. The method according to any one of claims 21-50 whereby albumin, immunoglobulins and .alpha.-1-antitrypsin are separated from plasma in less than 12 hours..Iaddend. .Iadd.56. The method according to any one of claims 21-50 whereby albumin, immunoglobulins and .alpha.-1-antitrypsin are separated from plasma in less than 6 hours..Iaddend. .Iadd.57. The method according to any one of claims 21-50 whereby the plasma is a pooled human plasma sample..Iaddend. .Iadd.58. The method according to claim 21, 26 or 30 whereby step (IV) is carried out after step (I)..Iaddend. .Iadd.59. A method for separating plasma components from a plasma sample by electrophoresis, comprising: (I) separating the plasma into a first and second component in a first electrophoretic separation apparatus using a first electrophoretic separation membrane having a molecular mass cut-off of about 200 kDa, whereby the first component containing a mixture of albumin and .alpha.-1-antitrypsin resides on one side of the first electrophoretic separation membrane while the second component resides on the other side of the first electrophoretic separation membrane; (II) removing undesired material from the first component in a second electrophoretic separation apparatus using a second electrophoretic separation membrane having a molecular mass cut-off of about 80 kDa, whereby albumin and .alpha.-1-antitrypsin reside on one side of the second electrophoretic separation membrane and the undesired material resides on the other side of the third electrophoretic separation membrane; (III) separating albumin from .alpha.-1-antitrypsin in the first component using a third separation membrane having a molecular mass cut-off of about 40 kDa in a third electrophoretic separation apparatus, whereby .alpha.-1-antitrypsin resides on one side of the third electrophoretic separation membrane and albumin resides on the other side of the third separation membrane; and (IV) separating the second component into third and fourth components whereby the third component containing immunoglobulins is located on one side of a fourth electrophoretic separation membrane, and the fourth component resides on the other side of the fourth separation membrane..Iaddend. .Iadd.60. The method according to claim 59, whereby the first, second, and third electrophoretic separation apparatus are connected together in steps (I)-(III), and the first electrophoretic separation is disconnected from the second or third electrophoretic separation apparatus in step (IV)..Iaddend.

Details for Patent RE39293

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Octapharma Pharmazeutika Produktionsges.m.b.h.	CUTAQUIG	immune globulin subcutaneous (human)-hipp	Solution	125668	12/12/2018		2040-01-28