Micro alternating tangential flow perfusion filter, micro bioreactor, and methods of use thereof

Patent metadata

Field	Value
Publication number	US-11946033-B2
Application number	US-202217729022-A
Country	US
Kind code	B2
Filing date	Apr 26, 2022
Priority date	Oct 26, 2017
Publication date	Apr 2, 2024
Grant date	Apr 2, 2024

Abstract

A micro alternating tangential flow (microATF) perfusion filter includes a hollow cylinder having a proximal end and a distal end. The proximal end is connected in series to a permeate chamber, followed by a retentate chamber. The proximal end either (i) terminates in or at the permeate chamber, or (ii) terminates in or at the retentate chamber. The portion of the proximal end within the permeate chamber, in a case of (ii), possesses at least one opening allowing fluid communication between an inside of the hollow cylinder and the permeate chamber. The microATF perfusion filter further includes an inlet, positioned over the retentate chamber, for communication with a source of positive or negative pressure, and an outlet, positioned in a wall of the permeate chamber, which can be connected to a check valve, which, in turn, can be connected to a hydrophobic fluid vent filter.

First claim

What is claimed is:

1. A micro alternating tangential flow perfusion device (microATF device) having dimensions suitable for an exchange volume of from 0.5-5 ml comprising
a hollow cylinder having a proximal end and a distal end, said proximal end being connected in series to a permeate chamber, followed by a retentate chamber, wherein the proximal end either (i) terminates in or at the permeate chamber, or (ii) terminates in or at the retentate chamber, wherein a portion of the proximal end is located within the permeate chamber, and in a case of (ii), possesses at least one opening allowing fluid communication between an inside of the hollow cylinder and the permeate chamber;
an inlet, positioned over the retentate chamber, for communication with a source of positive or negative pressure;
an outlet, positioned in a wall of the permeate chamber, which can be connected to a check valve, which, in turn, can be connected to a hydrophobic fluid vent filter.

2. The microATF device of claim 1 in which the outlet is capped with a check valve that allows gas to escape from the permeate chamber while allowing no gas to enter.

3. The microATF device of claim 1 in which the proximal end terminates in or at the permeate chamber.

4. The microATF device of claim 1 , which further comprises one or more hollow fiber filters each having a first end and a second end, the one or more hollow fiber filters arranged longitudinally inside the hollow cylinder with the first end extending in a direction of the distal end of the hollow cylinder and the second end terminating in or at the retentate chamber.

5. The microATF device of claim 4 in which the first end of the one or more hollow fiber filters extends down a length of the hollow cylinder and terminates at or near the distal end of the hollow cylinder.

6. The microATF device of claim 4 in which the one or more hollow fiber filters have an inside diameter ranging from about 0.5 mm to about 1 mm.

7. The microATF device of claim 4 in which the one or more hollow fiber filters comprises hollow fiber filter media having an average pore size ranging from about 0.2 μm to about 2.0 μm.

8. The microATF device of claim 4 in which the one or more hollow fiber filters have an average pore size of about 0.65 μm.

9. The microATF device of claim 4 in which the one or more hollow fiber filters have an average pore size that does not permit passage of molecules having a molecular weight ranging from about 20 kDa to about 70 kDa.

10. The microATF device of claim 9 in which the one or more hollow fiber filters have an average pore size that does not permit passage of molecules having a molecular weight of about 30 kDa or more.

11. A system comprising a housing having a volume of no more than about 15 ml equipped with a micro alternating tangential perfusion device (microATF device) having dimensions suitable for an exchange volume of from 0.5-5 ml comprising:
(a) a hollow cylinder having a proximal end and a distal end, said proximal end being connected in series to (i) a permeate chamber and (ii) a retentate chamber separated from the permeate chamber by a filter element, wherein the proximal end either (1) terminates in or at the permeate chamber, or (2) terminates in or at the retentate chamber, wherein a portion of the proximal end is located within the permeate chamber, and in a case of (2), possesses at least one opening allowing fluid communication between an inside of the hollow cylinder and the permeate chamber;
(b) an inlet, positioned over the retentate chamber, for communication with a source of alternating positive or negative pressure; and
(c) an outlet, positioned in a wall of the permeate chamber, which can be connected to a check valve, which, in turn, can be connected to a hydrophobic fluid vent filter;
and a fluid conduit connecting at least one well of the housing with the proximal end of the microATF device.

12. The system of claim 11 in which the housing is a multi-well plate with a capacity of 0.2 ml, 0.5 mL, 1.0 mL, 2.0 mL, 5.0 mL, 10 mL, or 15 mL.

13. The system of claim 11 in which the outlet is capped with a check valve that allows fluid to flow through the filter into the permeate chamber while preventing backflow through the filter into the retentate chamber.

14. The system of claim 11 , wherein the filter element comprises a tubular membrane or at least one hollow fiber membrane.

15. The system of claim 14 , where the filter element is characterized by a filter flux of 50-100 μL per cycle.

16. The system of claim 11 in which the housing holds about 10 mL, about 5 mL or less of a volume of cells in cell culture media.

17. The system of claim 13 in which the check valve is connected to a hydrophobic fluid vent filter.

18. The system of claim 12 , comprising a plurality of fluid conduits connecting a plurality of wells of the multi-well plate with a plurality of microATF devices.

19. The system of claim 11 in which the source of positive or negative pressure is a pipette.

20. The system of claim 11 in which the volume of cell culture media deposited into the permeate chamber ranges from about 50 μL to about 500 μL or from about 100 μL to about 300 μL.

Assignees

• Repligen Corp

Inventors

• Bransby Michael
• Yuen Philip

Classifications

• C12M29/10 Primary

  Perfusion · CPC title

• B01D61/147

  Microfiltration · CPC title

• B01D61/18

  Apparatus therefor · CPC title

• C12M29/04

  Filters; Permeable or porous membranes or plates, e.g. dialysis · CPC title

• C12M33/14

  with filters, sieves or membranes · CPC title