Tangential flow perfusion system

Patent metadata

Field	Value
Publication number	US-9663753-B2
Application number	US-201314430346-A
Country	US
Kind code	B2
Filing date	Sep 25, 2013
Priority date	Sep 27, 2012
Publication date	May 30, 2017
Grant date	May 30, 2017

Abstract

The invention discloses a system for perfusion culture of cells which comprises: a bioreactor; a filter unit with a retentate inlet end, a retentate outlet end and a permeate outlet port; a reciprocating pump fluidically connected to the retentate inlet end, with the retentate inlet end fluidically connected to the bioreactor via an inlet check valve arranged to allow flow in the direction from the bioreactor to the retentate inlet end and to block flow in the reverse direction, and where the retentate outlet end is fluidically connected to the bioreactor via an outlet check valve arranged to allow flow in the direction from the retentate outlet end to the bioreactor and to block flow in the reverse direction; and where the inlet and outlet check valves are each fluidically connected to a tubing branch point, which is further connected to the bioreactor via a length of tubing. In an alternative embodiment, each of said inlet and outlet check valves is fluidically connected to the bioreactor via separate lengths of tubing. The invention further discloses a pre-sterilized system for perfusion culture of cells which comprises: a bioreactor; a reciprocating pump and a filter unit fluidically connected to the reciprocating pump and fluidically connected to the bioreactor via at least one aseptic connector.

First claim

The invention claimed is:

1. A system for perfusion culture of cells comprising:
a) at least one bioreactor;
b) at least one filter unit comprising a retentate inlet end, a retentate outlet end that is on the opposite end of the at least one filter unit, and at least one permeate outlet port;
c) at least one reciprocating pump fluidically connected to the retentate inlet end,
wherein said retentate inlet end is fluidically connected to said at least one bioreactor via an inlet check valve arranged to allow flow in the direction from the at least one bioreactor to the retentate inlet end and to block flow in the reverse direction, and wherein said retentate outlet end is fluidically connected to said at least one bioreactor via an outlet check valve arranged to allow flow in the direction from the retentate outlet end to the at least one bioreactor and to block flow in the reverse direction; and
wherein said inlet and outlet check valves are each fluidically connected to a tubing branch point, which is further connected to the at least one bioreactor via a length of tubing.

2. The system according to claim 1 , wherein said length of tubing has an inner volume lower than or equal to 50% of the stroke volume of the at least one reciprocating pump.

3. The system according to claim 1 , further comprising an inlet control valve between said inlet check valve and said branch point.

4. The system according to claim 1 , further comprising an outlet control valve between said outlet check valve and said branch point.

5. The system according to claim 1 , further comprising an inlet pressure transducer arranged to measure the pressure at said retentate inlet end.

6. The system according to claim 1 , wherein the at least one reciprocating pump is a fluid-driven diaphragm pump and the system comprises a drive fluid pressure transducer arranged to measure a drive fluid pressure in the at least one reciprocating pump.

7. The system according to claim 5 , further comprising at least one control unit, arranged to control at least one of the at least one reciprocating pump, an inlet control valve, and an outlet control valve depending on pressure data received from said inlet pressure transducer.

8. The system according to claim 7 , wherein the at least one control unit is arranged to control at least one of the inlet control valve and the at least one reciprocating pump depending on pressure data received from said inlet pressure transducer.

9. The system according to claim 1 , further comprising a permeate pump and a permeate pressure transducer.

10. The system according to claim 1 , wherein said at least one filter unit is a hollow fiber cartridge or a flat sheet cassette.

11. The system according to claim 1 , wherein said at least one bioreactor comprises an inflatable flexible bag resting on a movable support, and wherein the length of tubing is a flexible length of tubing.

12. The system of claim 1 , wherein the at least one filter unit is fluidically connected to the bioreactor via at least one aseptic connector.

13. A method for perfusion culture of cells, comprising:
a) providing a system according to claim 1 ;
b) adding cell culture medium and cells to said at least one bioreactor;
c) cultivating cells in said at least one bioreactor under agitation;
d) operating, during cultivation, said at least one reciprocating pump to
i) withdraw fluid from said at least one bioreactor via said length of tubing, said branch point and said inlet check valve to said retentate inlet end of said at least one filter unit and to
ii) return fluid from said retentate outlet end of said at least one filter unit via said outlet check valve, said branch point and said length of tubing to said at least one bioreactor and;

e) withdrawing a permeate from at least one permeate outlet port.

14. The method according to claim 13 , further comprising adding at least one fluid to the at least one bioreactor during cultivation.

15. The method according to claim 13 , wherein the at least one filter unit comprises a microfiltration membrane and wherein the permeate comprises a target substance expressed by the cells.

16. The method according to claim 13 , wherein the at least one filter unit comprises an ultrafiltration membrane and wherein a target substance expressed by the cells is retained in the retentate and returned to the at least one bioreactor.

17. The method according to claim 13 , further comprising controlling at least one of the at least one reciprocating pump and an inlet control valve to keep the pressure reading from an inlet pressure transducer within a preset range during outward strokes of the at least one reciprocating pump.

18. The method according to claim 13 , further comprising calculating a transmembrane pressure from data provided by an inlet pressure transducer and an outlet pressure transducer; and controlling at least one of the at least one reciprocating pump and the inlet and outlet control valves to keep the transmembrane pressure between preset upper and lower limits.

19. The method according to claim 18 , wherein the trans-membrane pressure is kept within a first range during outward strokes of the at least one reciprocating pump and within a second range during inward strokes of the at least one reciprocating pump.

20. The method according to claim 13 , further comprising operating a permeate pump in a direction to produce a back-flush of a filter membrane.

21. A system for perfusion culture of cells comprising: a) at least one bioreactor; b) at least one filter unit comprising a retentate net end, a retentate outlet end that is on the opposite end of the at least one filter unit, and at least one permeate outlet port; wherein the at least one filter unit is fluidically connected to the at least one bioreactor via at least one aseptic connector, and c) at least one reciprocating pump fluidically connected to the retentate net end, wherein said retentate net end is fluidically connected to said at least one bioreactor via an net check valve arranged to allow flow in the direction from the at least one bioreactor to the retentate net end and to block flow in the reverse direction, and wherein said retentate outlet end is fluidically connected to said at least one bioreactor via an outlet check valve arranged to allow flow in the direction from the retentate outlet end to the at least one bioreactor and to block flow in the reverse direction; and wherein said net and outlet check valves are each fluidically connected to a tubing branch point, which is further connected to the at least one bioreactor via a length of tubing.

22. The system according to claim 21 , wherein said length of tubing has an inner volume lower than or equal to 50% of the stroke volume of the at least one reciprocating pump.

23. The system according to claim 21 , further comprising an inlet control valve between said inlet check valve and said branch point.

24. The system according to claim 21 , further comprising an outlet control valve between said outlet check valve and said branch point.

25. The system according to claim 21 , further comprising an inlet pressure transducer arranged to measure the pressure at said retentate inlet end.

26. The system according to claim 21 , wherein the at least one reciprocating pump is a fluid-driven diaphragm pump and the system comprises a drive fluid pressure transducer arranged to measure a dr

Assignees

• Ge Healthcare Bio Sciences Ab

Inventors

• Gebauer Klaus

Classifications

• B01D2313/243

  Pumps · CPC title

• C12M29/18

  External loop; Means for reintroduction of fermented biomass or liquid percolate (loop type reactors for chemical or physical processes B01J19/2435) · CPC title

• C12M29/02 Primary

  Percolation · CPC title

• C12M29/12

  Pulsatile flow · CPC title

• C12M29/16

  Hollow fibers (hollow fiber modules in general B01D63/02) · CPC title