Single use bioreactor

Patent metadata

Field	Value
Publication number	US-10801003-B2
Application number	US-201715613954-A
Country	US
Kind code	B2
Filing date	Jun 5, 2017
Priority date	Jun 3, 2016
Publication date	Oct 13, 2020
Grant date	Oct 13, 2020

Abstract

A single-use bioreactor is provided. The single-use bioreactor may include a bioprocess container, a shell, at least one agitator, at least one sparger, at least one gas filter inlet port for the sparger(s) and headspace overlay, at least one fill port, at least one harvest port, at least one sample port, and at least one probe. In examples, at least one controller may monitor and control one or more parameters associated with the single-use bioreactor A method to cultivate and propagate mammalian cells is also provided. The method may include cultivating under suitable conditions and in a suitable culture medium in a first single-use bioreactor, transferring the medium containing the cells obtained by propagation from the at least one mammalian cell is into a second single-use bioreactor, transferring the medium containing the cells obtained by propagation from the at least one mammalian cell is into a third single-use bioreactor, and cultivating the cells in the third bioreactor.

First claim

The invention claimed is:

1. A bioreactor comprising:
a bioprocess container made from a liquid impermeable and flexible shape-conforming material, the bioprocess container having a bottom and at least one side wall, the bioprocess container defining a hollow enclosure for receiving a culture media, the bioprocess container including an interior surface configured to contact a culture media contained in the hollow enclosure and an opposite exterior surface, wherein the flexible shape-conforming material comprises a polymer film, the polymer film having a modified surface that promotes homogeneous mixing of a culture media contained in the hollow enclosure, the modified surface defining the interior surface of the hollow enclosure, the hollow enclosure having a volume of from about 2 L to about 50,000 L;
a rigid shell defining an interior volume and having a shape configured to receive the bioprocess container therein, the rigid shell having a bottom portion defining an interior surface, the bioprocess container conforming to the interior surface of the bottom portion of the rigid shell; and
a mixing device comprising a rotatable shaft coupled to at least one impeller that extends into the hollow enclosure of the bioprocess container, the at least one impeller being made from a polymer material that has been modified to form a hydrophilic surface.

2. A bioreactor as defined in claim 1 , wherein the interior surface of the polymer film comprises a modified polyolefin.

3. A bioreactor as defined in claim 1 , wherein the interior surface of the polymer film comprises a modified low density polyethylene.

4. A bioreactor as defined in claim 1 , wherein the interior surface of the polymer film comprises acrylamide grafted onto a polyethylene, an oxidized polyethylene, or a polyethylene blend containing poly(2-hydroxyethyl methacrylate), poly(2,3-dihydroxypropyl methacrylate), or mixtures thereof.

5. A bioreactor as defined in claim 1 , wherein the interior surface of the polymer film comprises a polymer material that has been modified by grafting to the polymer material hydrophilic components using gamma, beta, or ultraviolet irradiation.

6. A bioreactor as defined in claim 1 , wherein the interior surface of the polymer film comprises a polymer material that has been modified by chemical oxidation.

7. A bioreactor as defined in claim 4 , wherein the low density polyethylene has been modified by being subjected to irradiation, photo induction, or oxidation.

8. A bioreactor as defined in claim 1 , wherein the impeller to bioprocess container diameter ratio is from about 0.35 to about 0.55.

9. A bioreactor as defined in claim 1 , wherein the bioreactor includes a top impeller and a bottom impeller and wherein the flow number (N q ) of the top impeller and the bottom impeller is from about 0.4 to about 0.9.

10. A bioreactor as defined in claim 1 , further comprising at least one baffle being configured to extend adjacent to the side wall of the bioprocess container in a longitudinal direction, the baffle having a shape that extends radially inward from the side wall an amount sufficient to affect fluid flow in the hollow enclosure during mixing of a culture media by the mixing device, the baffle being positioned between the interior surface of the rigid shell and the exterior surface of the bioprocess container.

11. A bioreactor as defined in claim 1 , wherein the bioreactor further comprises at least one sparger and wherein the sparger comprises a ballast sparger, the ballast sparger comprising a gas tube that has a longitudinal portion and a lateral portion, the longitudinal portion extending vertically into the hollow enclosure of the bioprocess container, the lateral portion being located at an end of the longitudinal portion below the impeller, the lateral portion defining a plurality of holes for releasing a gas into a culture media contained within the bioprocess container.

12. A bioreactor as defined in claim 1 , wherein the bioprocess container is in fluid communication with a drain line located at the bottom of the bioprocess container, and wherein a fluid collecting device is positioned between the hollow enclosure of the bioprocess container and the drain line, the fluid collecting device having a shape configured to induce a vortex flow of fluids from the bioprocess container into the drain line.

13. A bioreactor as defined in claim 1 , further comprising a controller in communication with at least one sensor, the controller being configured to receive information from the at least one sensor and, based on the information, to control a fluid supply for varying a flow rate of a fluid from the fluid supply into the hollow enclosure of the bioprocess container for maintaining at least one parameter of a culture media contained within the hollow enclosure within preset limits.

14. A bioreactor as defined in claim 13 , wherein the fluid supply comprises a carbon dioxide gas supply in fluid communication with the bioprocess container and a liquid alkali supply also in fluid communication with the bioprocess container, the at least one sensor comprising a pH sensor and wherein the controller is configured to regulate pH levels of a culture media within preset limits by adding amounts of carbon dioxide gas from the carbon dioxide gas supply for selectively lowering the pH or by adding amounts of an alkali from the liquid alkali supply for selectively increasing the pH.

15. A bioreactor as defined in claim 13 , wherein the fluid supply comprises an oxygen gas supply and wherein the at least one sensor comprises a dissolved oxygen sensor and wherein the controller regulates dissolved oxygen levels within a culture media within present limits by periodically adding amounts of oxygen gas from the oxygen gas supply to a culture media within the hollow enclosure of the bioprocess container based on information received from the dissolved oxygen sensor.

16. A bioreactor as defined in claim 13 , wherein the fluid supply comprises a carbon dioxide gas supply and wherein the at least one sensor comprises a dissolved carbon dioxide sensor and wherein the controller regulates dissolved carbon dioxide levels within a culture media within present limits by periodically adding amounts of carbon dioxide gas from the carbon dioxide gas supply to a culture media within the hollow enclosure of the bioprocess container based on information received from the dissolved carbon dioxide sensor.

17. A bioreactor as defined in claim 13 , further comprising a thermal jacket surrounding the bioprocess container, the thermal jacket being in fluid communication with at least one of a heated fluid or a chilled fluid, the bioreactor further comprising a temperature sensor for sensing a temperature of a culture media contained within the bioprocess container, the temperature sensor being in communication with the controller, and wherein the controller is configured to receive information from the temperature sensor and, based on the information, control flow of a fluid into the thermal jacket for increasing or decreasing the temperature of a culture media contained in the bioprocess container for maintaining a culture media within preset temperature limits.

18. A bioreactor as defined in claim 13 , wherein the at least one sensor comprises a pH sensor and a dissolved oxygen sensor that are both in communication with the controller and wherein the controller receives information from the pH sensor and the dissolved oxygen sensor and controls a flow of different fluids into the bioprocess container for maintaining pH

Assignees

• Lonza Ag

Inventors

• Jaques Colin Mark
• Khan Mohsan Waseem
• De Barros Costa Rita D'Ornelas P
• Beaney Anthony
• Valentine David

Classifications

• B01F27/86

  co-operating with deflectors or baffles fixed to the receptacle · CPC title

• B01F27/1134

  the impeller being of hydrofoil type · CPC title

• B01F35/513

  Flexible receptacles, e.g. bags supported by rigid containers · CPC title

• B01F27/053 Primary

  characterised by their materials · CPC title

• B01F23/231

  by bubbling (mixers with gas or liquid agitation, e.g. with air supply tubes B01F33/40) · CPC title