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EP-4741486-A1 - CULTIVATION SYSTEM AND CONTAINER ATTACHMENT FOR A CULTIVATION CONTAINER

EP4741486A1EP 4741486 A1EP4741486 A1EP 4741486A1EP-4741486-A1

Abstract

The invention relates to a container attachment which can be placed on the neck of a cultivation container to close the opening of the cultivation container, wherein the container attachment has an inner side and an outer side, the inner side facing the interior of the cultivation container and the outer side facing the exterior of the cultivation container when the container attachment is placed on the neck of the cultivation container, wherein the container attachment comprises at least one sensor unit and at least one dispensing unit, wherein the sensor unit is arranged at least partially on the inner side of the container attachment to enable parameter measurement inside the cultivation container, and wherein the dispensing unit is arranged at least partially on the inner side of the container attachment to enable liquid dispensing into the interior of the cultivation container. Further aspects of the invention relate to a cultivation system with such a container attachment and a method for cultivating biological material using such a cultivation system.

Inventors

  • OTT, CHRISTIAN
  • KOLBERG, CHRISTOPH

Assignees

  • SCHOTT AG

Dates

Publication Date
20260513
Application Date
20200626

Claims (15)

  1. Container attachment (100) for mounting on a cultivation container (50), preferably on a shake flask, wherein the cultivation container (50) has a neck (52) and an opening (54) extending through the neck, wherein the container attachment (100) comprises: an inner side (102) which faces the interior of the cultivation container (50) when the container attachment (100) is placed on the cultivation container (50) and an outer side (104) which faces the exterior of the cultivation container (50) when the container attachment (100) is placed on the cultivation container (50), characterized in that the container attachment (100) comprising at least one sensor unit (112), wherein the sensor unit (112) is arranged at least partially on the inside (102) of the container attachment (100) to enable parameter measurement inside the cultivation container (50), and at least one dispensing unit (122) with a reservoir (140) for liquid storage, wherein the dispensing unit (122) is arranged at least partially on the inside (102) of the container attachment (100) to enable liquid dispensing into the interior of the cultivation container (50).
  2. Container attachment (100) according to the preceding claim, wherein the container attachment (100) has an annular, in particular cylindrical, gap (106) for receiving a neck (52) of the cultivation container (50) and/or wherein the container attachment has an outer ring (105) which radially surrounds the neck (52) of the cultivation container (50) when the container attachment (100) is placed on the cultivation container (50) and/or wherein the container attachment (100) has an internal projection (103) which extends into the neck (52) of the cultivation container (50) when the container attachment (100) is placed on the cultivation container (50) and wherein the outer ring (105) and the inner projection (103) are preferably monolithic.
  3. Container attachment (100) according to one of the preceding claims, wherein the container attachment (100) can be placed vertically from above onto the cultivation container (50) and is preferably designed to be held on the cultivation container (50) by gravity.
  4. Container attachment (100) according to one of the preceding claims, wherein the container attachment (100) has a stop surface (108) located, in particular at the upper end of the gap (106), which comes into contact with the cultivation container (50) when the container attachment (100) is placed on the cultivation container (50) and wherein the container attachment (100) has a height measured from the stop surface of less than 10 centimeters, preferably less than 5 centimeters, more preferably less than 4 centimeters, more preferably less than 3 centimeters, particularly preferably less than 2 centimeters, and most preferably less than 14 centimeters.
  5. Container attachment (100) according to one of the preceding claims, wherein the reservoir (140) has an upper reservoir section (146) and a lower reservoir section (142), wherein the upper reservoir section (146) has a reflow opening (148) for the reflow of gas and wherein the lower reservoir section (142) is arranged on the inside (102) of the container attachment and has a discharge opening (144) to allow liquid to be discharged through the discharge opening (144) into the interior of the cultivation container (50).
  6. Container attachment (100) according to one of the preceding claims, wherein the reservoir (140) has an upper reservoir section (146) and a lower reservoir section (142), wherein the upper reservoir section (146) is wider than the lower reservoir section (142) and wherein the upper reservoir section (146) is preferably arranged on the outside (104) of the container extension (100) and is particularly preferably wider in order to to form a stop (147) when the dispensing unit (122) and/or the reservoir (140) is installed on a port of the container attachment (100).
  7. Container attachment (100) according to the preceding claim, wherein the container attachment (100) and/or the dispensing unit (122) has a connection between the inflow opening (148) of the upper reservoir section (146) and the inside of the container attachment (100) to allow gas to flow out of the interior of the cultivation container (50) when the container attachment (100) is placed on the cultivation container (50).
  8. Container attachment (100) according to one of the preceding claims, wherein the reservoir (140) has a refill opening (149) for refilling with liquid and wherein the container attachment (100) and/or the dispensing unit (122) preferably has a connection between the refill opening (149) of the reservoir (140) and a valve (164) arranged or arrangable on the outside of the container attachment (100) to allow refilling of liquid into the reservoir (140) and wherein the container attachment (100) and/or the dispensing unit (122) preferably has a holder (163) for the releasable fixing of the valve (164).
  9. Container attachment (100) according to one of the preceding claims, wherein the container attachment (100) has a connection between the inside (102) of the container attachment (100) and a valve (174) arranged or arrangable on the outside (104) of the container attachment (100) to allow addition or removal from the interior of the cultivation container (50) when the container attachment (100) is placed on the cultivation container (50) and wherein the container attachment (100) preferably has a holder (173) for the releasable fixing of the valve (174).
  10. Container attachment (100) according to one of the preceding claims, wherein at least one sensor unit (112) is designed as a biosensor unit for analyte-specific parameter measurement and/or wherein at least one or one further sensor unit (112) of the container attachment (100), which in particular is not designed as a biosensor unit, is designed as a luminophore unit for luminescence-based parameter measurement and/or wherein at least one or another sensor unit (112) of the container attachment (100) is designed as an alternating field unit for dielectric-based parameter measurement and/or wherein at least one or another sensor unit (112) of the container attachment (100) is designed as a transistor unit for field-effect-based parameter measurement and/or wherein the at least one sensor unit (112) is arranged or can be arranged on a front housing section of a sensor component (114) such that the at least one sensor unit (112) can be arranged on the inside of the container attachment (100) by installing the sensor component on a port (110) of the container attachment (100).
  11. Container attachment (100) according to one of the preceding claims, wherein the container attachment (100) comprises a plurality of sensor/dispensing units (112, 122), preferably at least one sensor unit (112) and at least two dispensing units (122).
  12. Cultivation system (10) encompassing: a container attachment (100) according to one of claims 1 to 11, as well as a cultivation container (50), in particular a handheld container, preferably designed as a shaker flask, for receiving a culture medium, wherein the cultivation container has a neck (52) and an opening (54) extending through the neck, wherein the container attachment (100) can be placed on the neck (52) of the cultivation container (50) in order to close the opening (54) of the cultivation container (50), in particular to close it sterilely, wherein the container attachment (100) has an inner side (102) and an outer side (104), wherein the inner side (102) faces the interior of the cultivation container (50) and the outer side (104) faces the exterior of the is facing the cultivation container (50) when the container attachment (100) is placed on the neck (52) of the cultivation container (50).
  13. Cultivation system (10) according to claim 12, wherein the cultivation container (50) has a flat bottom (60) for placement on a surface and/or wherein the neck (52) of the cultivation container (50) extends vertically upwards, particularly when the cultivation container (50) is placed on the ground, and/or wherein the container attachment (100) can be placed vertically from above onto the neck (52) of the cultivation container (50) and is preferably designed to be held on the neck (52) of the cultivation container (50) by gravity.
  14. Cultivation system (10) according to one of claims 12 or 13, wherein the cultivation vessel (50) has a volume of less than 2801 milliliters, preferably less than 1801 milliliters, more preferably less than 501 milliliters, again more preferably less than 251 milliliters, and particularly preferably less than 126 milliliters and/or wherein the cultivation container (50) has a diameter of less than 22 centimeters or less than 21 centimeters, preferably less than 14 centimeters or less than 12 centimeters, particularly preferably less than 9 centimeters or less than 8 centimeters and/or wherein the cultivation container (50) has a height of less than 31 centimeters, preferably less than 23 centimeters, more preferably less than 19 centimeters, particularly preferably less than 17 centimeters, and most preferably less than 14 centimeters.
  15. Method for propagating or cultivating biological material, preferably for the production of pharmaceuticals, in particular biopharmaceuticals, comprising: Providing a cultivation system (10) according to any one of claims 11 to 14, Providing a feed solution, in particular containing the substrates required for product formation, e.g. saccharides, wherein the feed solution is in the reservoir of the Delivery unit (122) is provided Providing a pH correction solution, in particular for raising the pH value, e.g. containing NaOH, wherein the pH correction solution is preferably provided in a further reservoir and/or is provided containing the feed solution, and wherein the sensor unit (112) records a parameter for process control in real time and the delivery unit (122) performs an active correction, e.g. pH correction or feed rate.

Description

The invention relates to a cultivation system with a container attachment for a cultivation container, in particular a handheld one, e.g. for a shaker flask. Bioreactors and shake flasks are used to cultivate microorganisms, animal and plant cells, thus opening up a wide range of applications for biotech production processes. There is a general need to further optimize these processes. In particular, improvements in product yield and thus profit increases are being driven for the production of biopharmaceuticals. Various approaches are available for controlling and monitoring production processes, increasing product yield, and reducing costs. Generally, processes can be controlled and monitored by determining substrate and product concentrations. However, this is time-consuming and usually requires resource-intensive offline analysis. Furthermore, the sampling process carries a significant risk of contamination. When using conventional bioreactors, process control and thus yield can be optimized by real-time process control of key parameters. In particular, in-situ monitoring of parameters such as temperature, metabolism- or product-formation-relevant substances, as well as real-time control of cultivation conditions, is advantageous for increasing product yield. In contrast, when shake flasks or other handheld cultivation containers are used, a sample draw is still required, which poses a contamination risk due to the handling involved with the cultivation unit open. Furthermore, the sample draw is time-consuming, as is the determination of substrate and product concentrations using offline analysis. A sample withdrawal can also mean that the interruption of mixing via a shaking incubator brings the mass exchange, especially of dissolved gases, to a standstill, inhibiting culture. Additionally, the multi-hour reaction time required for pH adjustments or substrate additions (feed), which is due to the duration of the off-line analysis, can be disruptive. Furthermore, the risk of contamination increases with each opening of the shake flask. Metabolism typically produces pH-lowering metabolic products. Attempts are sometimes made to counteract this by buffering the medium. Particularly during process development, pH monitoring and additions to adjust the pH may be necessary when the buffering capacity is exhausted. Medium components should not be introduced at arbitrarily high concentrations at the start of cultivation, as this can lead to undesirable substrate inhibition. To reduce the number of sample draws when using a shake flask, approaches include measuring the optical density and luminescence, e.g. for pH and pO2 , of glued-in spots through the shake flask. However, the optical quality of the shake flask wall (e.g., transmission, scattering) limits this principle. Furthermore, it must be considered that measuring cell growth via optical density requires a constant extinction coefficient. This coefficient, however, varies with the changing medium composition during cultivation. This results in a difference between viable and lysed cells. With embedded spots, the limitations of leaching and drift are further compounded. The DE202016000554U1 relates to a disposable connection device for insertion into a connection opening of a top plate of a bioreactor and/or into a connection opening of a bag bioreactor, with multiple passages and a fastening section, wherein the disposable connection device is made in one piece from plastic and the fastening section has a fastening structure on an outer circumferential surface for fastening the disposable connection device in the connection opening. The WO2014044612A1 relates to a reactor tank designed as a disposable element with a lid and/or opto-electronically readable sensor patches attached inside, a reactor comprising the reactor tank and a reactor tank receiving periphery, in turn comprising a reactor tank holder and optionally an opto-electronic measuring system for reading sensor patches, wherein the reactor tank holder is coupled to a drive unit for generating a rotary-oscillating movement of the reactor tank about its own central vertical axis, and the use of this device for cultivating cells and/or microorganisms. The EP3109314A1 relates to a cell cultivation device comprising a culture tank for receiving a culture solution containing a cell, a wave element that is at least partially arranged in the culture tank, a stirring mechanism that is supported by the wave element, is arranged in the culture tank and has at least one pair of stirring blades configured to be rotatable about the wave element, and a filter that is arranged in contact with the wave element to draw the culture solution from the culture tank and/or to supply the culture solution to the culture tank. The CN201658978U This relates to a biological experiment device comprising at least one bottle body; the bottle body is provided with a bottle opening; a bottle cork is arranged over the bottle