SYSTEMS AND METHODS FOR EXCHANGE OF BUFFER SOLUTIONS

• US 20170096448A1
• Filed: 05/21/2015
• Published: 04/06/2017
• Est. Priority Date: 05/21/2014
• Status: Active Grant

• Current Assignee: Unchained Labs Inc.
• Original Assignee: Unchained Labs Inc.
• Inventors: VARNI, John, LAMBERT, Stephen, BUSACCA, Robert, BURGE, Russell
• Granted Patent: US 10,640,531 B2
• Time in Patent Office: Days
• CPC Class Codes: B01D 2311/10 Temperature control
• CPC Class Codes: B01D 2311/14 Pressure control
• CPC Class Codes: B01D 2321/18 Use of gases
• CPC Class Codes: B01D 2321/2058 by vibration of the membrane, e.g. with an actuator
• CPC Class Codes: B01D 61/145 Ultrafiltration
• CPC Class Codes: B01D 61/18 Apparatus therefor
• CPC Class Codes: B01D 65/08 Prevention of membrane fouling or of concentration polarisation
• CPC Class Codes: C07K 1/34 by filtration, ultrafiltration or reverse osmosis
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/065 Purification, fragmentation
• CPC Class Codes: G01N 1/34 Purifying; Cleaning processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: G01N 1/40 Concentrating samples
• CPC Class Codes: G01N 2001/4088 filtration
• CPC Class Codes: G01N 2035/00485 combined with sample carriers
• CPC Class Codes: G01N 2035/1025 Fluid level sensing
• CPC Class Codes: G01N 35/1009 Characterised by arrangements for controlling the aspiration or dispense of liquids
• CPC Class Codes: G01N 35/1016 Control of the volume dispensed or introduced

Abstract

Systems and methods for exchanging buffer solutions are disclosed. In accordance with some embodiments, the methods and systems for buffer exchange may be automated and/or the methods and systems may include mixing during filtering operations.

Claims

1. An automated method for exchange of buffer solutions from admixtures comprising a buffer solution and a biological component, the method comprising:

providing a plurality of individual reservoirs containing an admixture comprising a biological component and a first buffer solution, the reservoirs containing a semi-permeable membrane;

pressurizing the reservoirs to force the first buffer solution through the semi-permeable membrane to produce a buffer-depleted residue;

detecting amounts of first buffer that was removed from the individual reservoirs; and

adding a second buffer to the reservoirs, an amount of second buffer added to the individual reservoirs being determined by the detected amount of first buffer that was removed the reservoir.

2. The method as set forth in claim 1 further comprising mixing the admixture to reduce fouling of the membrane.

3. The method as set forth in claim 2 wherein mixing is performed simultaneously with pressurizing the reservoirs to reduce fouling of the membrane while forcing the first buffer solution through the semi-permeable membrane to produce a buffer-depleted residue.

4. The method as set forth in claim 2 or claim 3 wherein mixing comprises vortexing the admixture.

5. The method as set forth in claim 4 wherein vortexing occurs in a direction normal to a flow direction to reduce fouling.

6. The method as set forth in any one of claims 1 to 5 comprising adding the second buffer in two or more cycles until a target exchange of first buffer is achieved.

7. The method as set forth in claim 3 wherein vortexing is controlled to maintain a constant flow rate through the semi-permeable membrane.

8. The method as set forth in claim 3 wherein vortexing is controlled to maintain a minimum flow rate through the semi-permeable membrane.

9. The method as set forth in any one of claims 1 to 8 wherein the amounts of first buffer that were removed from the individual reservoirs are detected by sensing a level of the admixture in the reservoir.

10. The method as set forth in any one of claims 1 to 9 wherein the amounts of first buffer that were removed from the individual reservoirs are detected by acoustic sensing, capacitance, light reflectance, pressurization time or weight measurement.

11. The method as set forth in any one of claims 1 to 10 wherein the detected amount of first buffer that was removed from the individual reservoirs is a detected volume.

12. The method as set forth in any one of claims 1 to 11 wherein the biological component is selected from the group consisting of proteins, peptides, antigens, antibodies, enzymes, microorganisms DNA and RNA.

13. The method as set forth in any one of claims 1 to 12 wherein the biological component is a protein and has a molecular weight of greater than about 20 kDa.

14. The method as set forth in any one of claims 1 to 13 further comprising sending a signal relating to the detected amount of first buffer that was removed from the individual reservoirs to a control system operable to control a dispenser system for dispensing the second buffer solution into the reservoirs.

15. The method as set forth in claim 14 wherein the second buffer solution is added the reservoirs while the first buffer solution is withdrawn the reservoirs.

16. The method as set forth in any one of claims 1 to 15 wherein the semi-permeable membrane is characterized by a molecular weight cut off of about 100 kDa or less.

17. The method as set forth in any one of claims 1 to 15 wherein the semi-permeable membrane is characterized by a molecular weight cut off of about 10 kDa or less.

18. The method as set forth in any one of claims 1 to 17 wherein each reservoir has a working volume of about 75 ml or less, about 25 ml or less, about 16 ml or less, about 8 ml or less, about 4 ml or less, about 1 ml or less, about 750 μ

l or less, about 500 μ

l or less or about 250 μ

l or less.

19. The method as set forth in any one of claims 1 to 18 wherein at least 3, at least 5, at least 10, at least 12, at least 16, at least 48 or at least 96 reservoirs are simultaneously buffer exchanged.

20. The method as set forth in any one of claims 1 to 19 comprising controlling the temperature of the admixture during buffer exchange.

21. The method as set forth in any one of claims 1 to 20 wherein the buffer solution further comprises an excipient.

22. The method as set forth in any one of claims 1 to 21 wherein the reservoirs are mounted on a single substrate.

23. The method as set forth in any one of claims 1 to 22 wherein the volumetric ratio of the first buffer removed from each reservoir and the second buffer added to the reservoirs is about 1:

1.

24. The method as set forth in any one of claims 1 to 22 wherein the volumetric ratio of the first buffer removed from each reservoir and the second buffer added to the reservoirs is less than 1:

1 to dilute the biological component or greater than 1;

1 to concentration the biological component.

25. A system for automated exchange of buffer solutions from admixtures comprising a buffer solution and a biological component, the system comprising:

a pressure chamber for receiving a plurality of reservoirs having a semi-permeable membrane and for creating a pressure difference across the membrane to force a first buffer solution through the membrane and produce a first buffer-depleted residue in the reservoir;

a sensor for detecting the level of fluid in the reservoirs; and

a dispensing system for adding a second buffer solution to the reservoirs, the dispensing system being configured to add an amount of second buffer to the reservoirs based on the sensed level of the buffer-depleted residue in the reservoirs.

26. The system as set forth in claim 25 further comprising a mixer for mixing the admixture while removing first buffer solution from the admixture.

27. The system as set forth in claim 26 wherein the mixer is a vortex mixer.

28. The system as set forth in any one of claims 25 to 27 further comprising a waste container for collecting filtrate forced through the semi-permeable membranes.

29. The system as set forth in any one of claims 25 to 28 further comprising a temperature control device for controlling the temperature of the admixtures.

30. The system as set forth in any one of claims 25 to 29 further comprising a plurality of buffer source containers.

31. The system as set forth in any one of claims 25 to 30 further comprising a control system operable to control a dispenser for dispensing the second buffer solution into the reservoirs based on a signal from the sensor.

32. The system as set forth in any one of claims 25 to 31 wherein the sensor is operable for acoustic sensing, capacitance sensing, light reflectance, time for pressurization, or weight measurement.

33. The system as set forth in any one of claims 25 to 32 in combination with the reservoirs, the reservoirs comprising a semi-permeable membrane characterized by a molecular weight cut off of about 100 kDa or less.

34. The system as set forth in any one of claims 25 to 32 in combination with the reservoirs, the reservoirs comprising a semi-permeable membrane characterized by a molecular weight cut off of about 10 kDa or less.

35. The system as set forth in claim 33 or claim 34 wherein each reservoir has a volume of about 75 ml or less, about 25 ml or less, about 16 ml or less, about 8 ml or less, about 4 ml or less, about 1 ml or less, about 750 μ

l or less, about 500 μ

l or less or about 250 μ

l or less.

36. The system as set forth in any one of claims 33 to 35 comprising at least 2, at least 3, at least 5, at least 10, at least 12, at least 16, at least 48 or at least about 96 reservoirs.

37. The system as set forth in any one of claims 25 to 36 further comprising a receiver plate having at least 2, at least 3, at least 5, at least 10, at least 12, at least 16, at least 48 or at least about 96 openings to receive reservoirs.

38. A method for removal of a low molecular weight carrier from an admixture comprising a high molecular weight component or microorganism and the low molecular weight carrier, the method comprising:

providing a plurality of reservoirs containing an admixture comprising a high molecular weight component and low molecular weight carrier, the reservoirs containing a semi-permeable membrane;

pressurizing the reservoirs to force the low molecular weight carrier through the semi-permeable membrane to produce a carrier-depleted residue; and

mixing the admixture while pressurizing the reservoirs to remove build-up of residue at a surface of the semi-permeable membrane.

39. The method as set forth in claim 38 wherein the admixture is vortexed while pressurizing the reservoirs to remove build-up of residue at a surface of the semi-permeable membrane.

40. The method as set forth in claim 39 wherein vortexing occurs in a direction normal to a flow direction to reduce fouling.

41. The method as set forth in any one of claims 38 to 40 wherein the high-molecular weight component is selected from the group consisting of proteins, peptides, antigens, antibodies, enzymes, microorganisms DNA and RNA.

42. The method as set forth in any one of claims 38 to 41 wherein the semi-permeable membrane is characterized by a molecular weight cut off of about 100 kDa or less.

43. The method as set forth in any one of claims 38 to 41 wherein the semi-permeable membrane is characterized by a molecular weight cut off of about 10 kDa or less.

44. The method as set forth in any one of claims 38 to 43 wherein each reservoir has a working volume of about 75 ml or less, about 25 ml or less, about 16 ml or less, about 8 ml or less, about 4 ml or less, about 1 ml or less, about 750 μ

l or less, about 500 μ

l or less or about 250 μ

l or less.

45. The method as set forth in any one of claims 38 to 44 wherein at least 3, at least 5, at least 10, at least 12, at least 16, at least 48 or at least 96 reservoirs are simultaneously buffer exchanged.

46. The method as set forth in any one of claims 38 to 45 wherein the reservoirs are mounted on a single substrate.

47. A system for removing a low molecular weight carrier from an admixture comprising a high molecular weight component or microorganism and the low molecular weight carrier, the system comprising:

a pressure chamber for receiving a plurality of reservoirs having a semi-permeable membrane and for creating a pressure difference across the membrane to force the low molecular weight carrier through the membrane and produce a carrier-depleted residue in the reservoir; and

a mixer for mixing the admixtures while removing carrier from the admixture.

48. The system as set forth in claim 47 wherein the mixer is a vortex mixer for vortexing the admixtures while removing carrier from the admixture.

49. The system as set forth in claim 47 or claim 48 further comprising a waste container for collecting filtrate forced through the semi-permeable membranes.

50. The system as set forth in any one of claims 47 to 49 in combination with the reservoirs, the reservoirs comprising a semi-permeable membrane characterized by a molecular weight cut off of about 100 kDa or less.

51. The system as set forth in any one of claims 47 to 49 in combination with the reservoirs, the reservoirs comprising a semi-permeable membrane characterized by a molecular weight cut off of about 10 kDa or less.

52. The system as set forth in claim 50 or claim 51 wherein each reservoir has a volume of about 75 ml or less, about 25 ml or less, about 16 ml or less, about 8 ml or less, about 4 ml or less, about 1 ml or less, about 750 μ

l or less, about 500 μ

l or less or about 250 μ

l or less.

53. The system as set forth in any one of claims 50 to 52 comprising at least 2, at least 3, at least 5, at least 10, at least 12, at least 16, at least 48 or at least about 96 reservoirs.

54. The system as set forth in any one of claims 47 to 53 further comprising a receiver plate having at least 2, at least 3, at least 5, at least 10, at least 12, at least 16, at least 48 or at least about 96 openings to receive reservoirs.