METHOD AND DEVICE ASSEMBLY FOR PROVIDING A LARGE AMOUNT OF CELL CULTURE MEDIUM USING DISPOSABLE COMPONENTS

Abstract

A method for providing a large amount of cell culture medium (10), in particular for cultivating animal cells, for a large reactor system (12) by using single-use components, comprises the following sequence of steps: a) preparing a concentrated media stock solution (32), b) filling the concentrated media stock solution (32) into single-use containers (40), c) transporting the single-use containers (40) filled with media stock solution (32) to the large reactor system (12), d) aseptically connecting the single-use containers (40) to the large reactor system (12) by means of a single-use hose system (26), e) processing the media stock solution (32) by supplying ultrapure water (24) and/or additives (44), and f) supplying the processed media stock solution (32) into the large reactor system (12).

Claims

1. A method for providing a large amount of cell culture medium for a large reactor system by using single-use components, comprising a sequence of steps as follows: a) preparing a concentrated media stock solution, b) filling single-use containers with the concentrated media stock solution, c) transporting the single-use containers filled with the media stock solution to the large reactor system, d) aseptically connecting the single-use containers to the large reactor system by means of a single-use hose system, e) processing the media stock solution by supplying ultrapure water and/or additives, and f) supplying the processed media stock solution into the large reactor system.

2. The method according to claim 1, characterized in that in step a) the concentrated media stock solution is matched to the volumes of subsequent mixing systems and/or to the reactor volume.

3. The method according to claim 1, characterized in that in step a) the concentrated media stock solution has a concentration of 5:1 or higher.

4. The method according to claim 1, characterized in that in step b) transportable single-use containers are filled with the concentrated media stock solution wherein the transportable single-use containers each having a volume of 1 L to 500 L.

5. The method according to claim 1, characterized in that between step a) and step b) the following steps are carried out: g) filling the concentrated media stock solution into single-use containers, and h) preparing the media stock solution by supplying ultrapure water and/or additives.

6. The method according to claim 1, characterized in that in step e) unstable chemical components are supplied to the media stock solution.

7. The method according to claim 1, characterized in that the single-use hose system comprises a static mixer which dilutes the media stock solution, to obtain a lower concentration.

8. The method according to claim 1, characterized in that the media stock solution is sterile-filtered before step f).

9. The method according to claim 1, characterized in that in the single-use hose system sensors are arranged, which are used for controlling at least one of the steps e) and f) by means of an automation unit.

10. An apparatus arrangement for providing a large amount of cell culture medium, characterized in that the apparatus arrangement is configured to carry out the method according to claim 1.

11. The apparatus arrangement according to claim 10, characterized by an electronic monitoring and control device, which is configured to adjust the concentration of the concentrated media stock solution in step a).

12. The apparatus arrangement according to claim 11, characterized by sensors for determining the pH value and/or the conductivity of the concentrated media stock solution, wherein the monitoring and control device is configured to control the adjustment of the concentration of the concentrated media stock solution on the basis of data of the sensors.

13. The apparatus arrangement according to claim 10, characterized by an automation unit for automated control of the processing of the media stock solution and/or supply of the processed media stock solution into the large reactor system.

14. The apparatus arrangement according to claim 10, characterized by a single-use hose system with aseptic connections, and a static mixer which dilutes the media stock solution.

15. The apparatus arrangement according to claim 14, characterized by a sensor for determining flow rate.

16. The apparatus arrangement according to claim 10, characterized in that the single-use containers can be sterilized by gamma radiation.

17. The apparatus arrangement according to claim 10, characterized in that the reactor system comprises a single-use bioreactor made of a suitable plastic material.

18. The apparatus arrangement according to claim 10, characterized in that the reactor system comprises a reactor made of stainless steel.

19. The method for providing a large amount of cell culture medium to a large reactor system for cultivating animal cells.

Description

[0032] Further advantages and features can be taken from the following description in conjunction with the attached drawing. The only FIGURE shows a schematic representation of the used appliances and the sequence of the method according to the invention.

[0033] The FIGURE schematically shows the apparatus arrangement according to the invention and the method according to the invention, by means of which a large amount of cell culture medium 10 in the form of a stock solution is provided in a large reactor system 12.

[0034] The large reactor system 12 comprises a reactor 14 and is part of a reactor installation 16, which furthermore comprises an automation unit 18, a sterile filtration device 20 as well as a WFI (Water for Injection) installation 22, which provides ultrapure water 24. The reactor system 12, the automation unit 18 and the WFI installation 22 are connected to each other via a single-use hose system 26 which includes a static mixer 28 and a plurality of sensors 30.

[0035] The sensors 30 in particular are provided for determining the flow rates, the pH values and the conductivity. The pressure determination also is of interest, above all with regard to a safety shut-off. Furthermore, sensors 30 can be provided for optical spectroscopic identification methods and quantitative analysis methods.

[0036] The reactor 14 is a stainless steel reactor with an operating volume of 10,000 liters, which in contrast to the single-use components employed in the method is further utilized for subsequent productions. The reactor installation 16 therefore is a hybrid installation.

[0037] In a first step of the method according to the invention a concentrated media stock solution 32 is produced outside the reactor installation 16, for example at a media manufacturer. The concentrated media stock solution 32 is obtained from powdery and/or liquid components 34 which are provided in tubular bags 36.

[0038] The concentrated media stock solution 32 can be produced in a single-use mixing system (not shown).

[0039] The concentration of the concentrated media stock solution 32 is adjusted via a SCADA (Supervisory Control and Data Acquisition) system 38 and controlled by means of sensor data of the pH value and the conductivity. The concentrated media stock solution 32 is produced with a particularly high concentration of 10:1, i.e. each unit volume of the media stock solution 32 contains only very little water.

[0040] In a next step the concentrated media stock solution 32 is filled into sterile single-use containers 40. The single-use containers 40 each hold a volume of about 200 liters and can be transported by a person using a trolley 42. Furthermore, the single-use containers 40 can be sterilized by gamma radiation, whereby they can be sterilized in a simple way before filling.

[0041] Subsequently, the single-use containers 40 filled with concentrated media stock solution 32 are temporarily stored, before they are transported to the reactor system 12.

[0042] In an optional step the concentrated media stock solution 32, in particular shortly before the transport to the reactor system 12, is diluted further and/or further additives are admixed to the concentrated media stock solution 32 in order to comply with the individual requirements of the reactor system 12 or the user of the media stock solution 32. The additives in particular can include constituents with a limited durability or useful life, which is why they are not suitable for a temporary storage and therefore are supplied shortly before the transport.

[0043] Alternatively, further additives can also be admixed to the concentrated media stock solution 32 directly during the production, in particular when the properties of the concentrated media stock solution 32 do not change disadvantageously until being supplied into the reactor system.

[0044] After this preparation of the concentrated media stock solution 32, the same is again filled into corresponding sterilized single-use containers 40.

[0045] During the production in the first step or in the optional step during the preparation of the media stock solution 32, the concentration of the concentrated media stock solution 32 is matched to the volume of the reactor 14 and of the automation unit 18 so that the required amount of cell culture medium 10 can be provided by the concentrated media stock solution 32 without any modification of the reactor installation 16.

[0046] After filling the produced or prepared concentrated media stock solution 32 and after the optional temporary storage, the single-use containers 40 filled with the media stock solution 32 are transported to the reactor system 12.

[0047] In a subsequent step, the single-use containers 40 are aseptically connected to the reactor system 12 by means of the single-use hose system 26.

[0048] The media stock solution 32 is processed in a next step. The media stock solution 32 here is diluted with ultrapure water 24 from the connected WFI installation 22 by means of the static mixer 28 to obtain a 1:1 concentration. In addition, additives 44 are metered to the diluted media stock solution 32 via the automation unit 18. These additives 44 in particular contain unstable chemical components which due to their short useful life are admixed to the media stock solution 32 only at this time in order to be able to ensure a particular concentration of these additives 44 in the reactor system 12.

[0049] Of course, in this step less critical additives 44 can also be supplied to the media stock solution 32 or additives 44 which are available only to the user of the media stock solution 32, for example due to licences or safety regulations.

[0050] Subsequently, the processed media stock solution 32 in a further step is introduced into the sterile filtration device 20 and sterile-filtered.

[0051] In the last step, the sterile-filtered processed media stock solution 32 is supplied to the reactor system 12 and now is available there for example for the cultivation of animal cells.

[0052] The automation unit 18 monitors all the aforementioned steps which take place within the reactor installation 16 and controls the entire process by means of the sensor data which are transmitted to the automation unit 18 by the sensors 30. Thus, a high process safety is ensured.

[0053] In this way, the method according to the invention allows to provide a large amount of stock solution in a large reactor system 12, which in particular is part of a hybrid installation, and at the same time to reduce the transport expenditure and the risk of a contamination of the reactor system 12. In the exemplary embodiment described above 10,000 liters of cell culture medium are provided by the method of the invention by means of fewer single-use containers each with a capacity of only 200 liters.