WO2022106006A1

WO2022106006A1 - Fumigation of an apparatus for handling cell cultures

Info

Publication number: WO2022106006A1
Application number: PCT/EP2020/082725
Authority: WO
Inventors: Patrick Clemente GILLIGAN; Kenneth B. K. Teo; David Anthony BULLINARIA; Neil David WALCUCH
Original assignee: Aixinno Ltd.
Priority date: 2020-11-19
Filing date: 2020-11-19
Publication date: 2022-05-27

Abstract

Apparatus for handling cell cultures comprising a chamber (2) being arranged in a container (1) and means for providing a flow of a fumigant through at least one region of said chamber (2), wherein said means comprising a fumigant generator (14) being connected with an inlet conduit (4) to an inlet section (7) of said chamber (2), wherein at least one first gas flow generator (3) is provided for generating a laminar flow of said fumigant or of air through said at least one region. A second gas flow generator (6) is provided for generating a turbulent gas flow in the at least one region. The invention further relates to a method for fumigation of at least one region of such a chamber. The laminar gas flow can be disturbed or a second turbular gas flow can be generated.

Description

Fumigation of an apparatus for handling cell cultures

Technical Field

[0001] The invention relates to an apparatus for handling cell cultures. Such an apparatus comprises at least one chamber, wherein the chamber may be part of a module, which is sealed from an external environment. Modules can be store modules for storing cell cultures or a lab ware. The chamber is arranged inside of a container. The apparatus comprises means for providing a flow of a fumigant through at least one region of said chamber. The fumigant is generated by a fumigant generator connected with an inlet conduit to an inlet section of the chamber. [0002] The invention further relates to a method for fumigation of at least one region of a chamber preferably of an apparatus as mentioned above.

Prior Art

[0003] CN 104498359 A discloses an integrated cell culture machine comprising means for sterilisation of at least parts of a chamber of the device.

[0004] US 8,741,228 B2 discloses an apparatus for decontamination of a region of a chamber, wherein a sterilisation gas is fed into the chamber forming a first flow through the chamber and leaving the chamber through an outlet section.

[0005] DE 19954 550 Al discloses a device for sterilisation of a chamber of a container. A flow generator generates a turbular gas flow inside the chamber. [0006] US 7,132,083 B2 discloses a mobile device for removing a sterilant from a room containing a sterilant atmosphere.

[0007] WO 2009/ 037231 Al discloses hydrogen peroxide for sterilisation of a cabinet.

[0008] WO 2020/098960 Al discloses a system for processing biological material comprising several modules, wherein one of the modules is a process module, and other modules forming storage devices for cell cultures, liquids, microplates or lab ware. The modules are sealed from the external environment and from adjacent modules. Lab ware or other material can be passed from one module to another without being exposed to contamination from the external atmosphere. The process module contains equipment, such as a liquid handling apparatus, pipettes, delidders, grippers and decaper. Typically, material for a workflow is loaded from one of the stores into the process module to be processed. After processing, the material is transported to a storage module. The process module is ready for the next process. The process module may have a working volume of about half a cubic meter.

[0009] It is desirable to avoid that one material is contaminated by a different material after processing different cell cultures. In some cases, it is desirable to strictly avoid that the material from one process contaminates the material from the next process. It is to be avoided that material of a therapeutic for one patient, contaminates material of a therapeutic for a second patient. Decontamination between two different materials may be achieved by fumigation with hydrogen peroxide. Hydrogen peroxide can kill cells and destroy biomolecules, such as lipids, proteins, DNA and RNA. The mechanism presumably results from oxidative damage, as hydrogen peroxide is highly oxidising. [0010] A typical time for a state-of-the-art process for fumigation of at least one region of such a chamber can be 5 to 10 minutes. Usually, the cycle time for hydrogen peroxide fumigation is around 8 hours (although it can be as short as 30 to 40 minutes). Even at 30 to 40 minute cycle time, the process module could spend much more time on fumigation than on productive work, generally limiting throughput.

[0011] Devices of the state-of-the-art may use a laminar flow of fumigant. A laminar flow generates boundary layer at surfaces. The fumigant has to diffuse through such a boundary layer. A laminar flow further comprises the disadvantage of producing stationary vortices trapping gas or not reaching each part of the volume.

[0012] Similar problems arise during purge step, wherein air is flowed through the volume.

Summary of the Invention

[0013] An object of the invention is to accelerate a fumigation of a chamber of a container. A further object of the invention is to improve an apparatus for handling cell cultures or a method for handling cell cultures or a method for fumigation of a chamber.

[0014] The object is attained with inventions specified in the claims, wherein each claim basically represents an independent solution of the above defined object.

[0015] An essential feature of the invention is providing a second flow of fumigant or air through the at least one region of the chamber of the container. The apparatus of the invention comprises a first gas flow generator, wherein the first gas flow generator generates a first gas flow inside the chamber. The first gas flow is preferably a laminar gas flow through the chamber from an inlet section to an outlet section. The outlet section may form a plenum.

[0016] There is a fumigant generator, which may be a gas generator providing the fumigant which may be hydrogen peroxide. There may be an inlet conduit for flowing the fumigant from the fumigant generator to the inlet section of the chamber. The flow generator being realised by a fan may be located in the inlet section of the chamber. The flow generator produces the first flow which is a laminar flow. A filter like a HEPA filter may be located downstream of the flow generator. One or more fans of the first flow generator drive a gas flow of a mixture of sterilisation gas e.g. hydrogen peroxide and a carrier gas, for example air, through the filter. The filter may have a function of a diffusor forming a gas flow with a homogeneous flow velocity. The second flow generator comprising at least one fan generates a second gas flow. During a purge step air is flowed through the volume of the chamber.

[0017] The second flow generator can modify the laminar gas flow or disturb the laminar gas flow to generate a turbulent gas flow. Preferably, the second flow generator generates a second gas flow, which is an independent gas flow with respect to the first gas flow. The second flow generator produces a gas flow, which may be perpendicular to the first gas flow. The angle between first gas flow direction and second gas flow direction may be between 60 and 120°. The first laminar gas flow may define an axis, wherein the axis runs between an inlet section or an upper section of the chamber to an outlet section or a lower section of the chamber. The axis can be a vertical axis. The second flow generator generates a flow around the axis. The flow generated by the second flow generator may be a circular flow or a vortex. The first laminar gas flow may be a slow flow. The flow velocity of the laminar gas flow may have values less than 0.3 to 0.5 m/s. The flow velocity of the turbulent flow may be at least two times, five times or ten times as high. There may be a heat generator for heating the fumigant. The heat generator may be located downstream of the fumigant generator and upstream of the inlet section or in the inlet conduit. The heat generator forms a heater for heating the fumigant, which is preferably a mixture of a sterilisation gas with air to an elevated temperature. The outlet section of the chamber is connected to an outlet conduit. The outlet section may be connected via a conduit to the inlet section to generate a circular flow of the fumigant. The circular flow may pass the heater and an optional pump or fan to convey the gas mixture.

[0018] The before mentioned two flow generators are used to generate a laminar and a turbulent fumigant flow. The flow generators may further be used for purging the chamber after a fumigation step. During the purge step a laminar flow of air and a turbulent flow of air is generated.

[0019] A further feature of the invention is alternating the flow pattern inside the chamber or at least inside one region of the chamber during the fumigation step and/ or during the purge step. In a first step the fumigant and preferably the heated fumigant is flowed in a laminar manner through the chamber or said region of the chamber. The first step may have a duration of 5 to 30 seconds. In a second step which preferably immediately follows the first step the fumigant and preferably the heated fumigant is flowed in a turbulent manner through the chamber or said region of the chamber. The second step may have a duration of 5 to 30 seconds. A sequence each of a first step and a second step may be repeated for several times. The sequence can be repeated for the time required for the fumigation, for example 5 to 10 minutes. After completing the fumigation the fumigant can be passed through a catalytic scrubber. The catalytic scrubber degenerates the sterilisation gas. Hydrogen peroxide react into oxygen and water. Ultraviolet light can be used to degenerate the fumigation gas. There may be a sensor downstream of the scrubber to detect residual sterilisation gas. In a similar manner the volume of the container is purged after the fumigation step. In a sequence a laminar flow and a turbulent flow of air can be generated and flowed through the chamber.

[0020] An apparatus or a method of the invention comprises especially one of the following features:

[0021] In the chamber, fumigation is accelerated by a combination of heating, and alternating between laminar flow for efficient gas exchange, and turbulent mixing.

[0022] Heating the chamber, or the incoming gas mixture, which contains the H2O2 vapour, will accelerate H2O2 destruction or neutralisation of biological material in the process module.

[0023] The chamber is provided with fans for driving air flow, an inlet HEPA filter, and a plenum for receiving the air. The chamber may further be provided with a duct to recirculate air from the plenum to the inlet HEPA filter. If the airflow driven by the fans is relatively slow (less than 0.3 to 0.5 meters per second (m/ s)), then the air flow can be laminar, which gives efficient air exchange.

[0024] The chamber is further provided with an H2O2 vapour generator, and e.g. an outlet or catalytic scrubber for H2O2. UV LEDs may further be provided to further accelerate H2O2 destruction. There is an H2O2 sensor provided to monitor H2O2 concentration in the outlet, and a valve provided to isolate the sensor from the initial, high concentrations of H2O2, which may be damaging to the sensor. Valves are provided as necessary to efficiently control the flow of gas.

[0025] Laminar flow enables efficient gas exchange, however, during laminar flow, a 'boundary layer⁷ of air tends to get trapped on surfaces (e.g. of equipment), and the exposure of the surfaces, e.g., to H2O2 during disinfection or air during purging, therefore relies on diffusion through the boundary layer, which is relatively slow. Also, while "dead spots" can be minimised in a laminar flow regime, air or fumigant may be trapped in cavities in and around equipment (similar to parasitic drag on an aircraft), and this air will also be exchanged slowly in laminar flow regimes.

[0026] There is also provided at least a first fan, which can be switched on intermittently, to create a fast turbulent flow, orthogonal to the laminar flow. For instance, the fan may be positioned to create a vortex about the vertical axis of the process module. The fast, turbulent flow will favor mixing, and will tend to disrupt the boundary layer of air or fumigant on various surfaces, thereby greatly increasing the exposure of those surfaces to H2O2 vapour or air. Furthermore, since the direction of fast, turbulent airflow is orthogonal to the direction of the laminar flow, it will tend to mix pockets of air or fumigant that are shielded from the laminar flow.

[0027] When it is desired to fumigate the chamber, H2O2 vapour is heated and pumped into the chamber. The H2O2 vapour can be recirculated in order to increase the efficiency of gas exchange, and also to fumigate the HEPA filter, plenum (outlet section), and recirculation duct. In order to increase the efficiency, the air flow is alternated between laminar flow, and fast turbulent mixing. For instance, the air may be circulated in a laminar flow regime, less than 0.3 - 0.5 m/s, using the large fans above the HEPA filter. This is alternated with fast, turbulent flow, by turning on the other fan, in the chamber. For instance, the flow may be maintained as laminar for 30 seconds, then switched to fast turbulent mixing, for 10 seconds. This is repeated for the time required for fumigation, e.g. 5 to 10 minutes.

[0028] Once the time required for fumigation is finished, H2O2 is purged from the chamber. The H2O2 may be purged via an external vent, or may be purged by recirculating it through e.g. a catalytic scrubber. While the air is being recirculated, the airflow is once again alternated between laminar flow (e.g. for 5 to 30 seconds per cycle) and fast turbulent mixing (e.g. for 10 seconds per cycle), until the H2O2 is purged down to a desired limit. The H2O2 concentration may be monitored on the outlet. The H2O2 sensor may be sensitive to damage resulting from high concentrations of H2O2, so the sensor may be protected by a valve, until the H2O2 concentration has reduced far enough not to damage the sensor. Once the H2O2 concentration has fallen to the predetermined limit in the outlet, the purge may be completed, and the next process begun.

[0029] A second fan for generating the turbulent flow of fumigant or air may be located inside the chamber or may be located outside the chamber in a separate housing. There is a flow connection between the separate housing and the chamber wall, which may form a nozzle or a baffle. The fan can be used to create a slight negative pressure in the chamber to drag air or fumigant out of the chamber. Valves can be provided in a duct between the chamber and the fan. Additionally, a filter, in particular an HEPA filter, can be located in flow direction upstream of an outlet fan. In a different embodiment of the invention the second fan generates a gas flow through a channel, which is connected to a chamber by a number of nozzles. There may be several nozzles displaced from each other in a vertical direction feeding air or fumigant at different levels into the chamber to form a turbulent flow pattern in the chamber. It is further possi- ble to realise the fumigant generator and/ or the scrubber by a separate module, which can be connected with a detachable pipe connection to a gas supply system, if needed.

Brief description of the Drawings

[0030] These and other features of the present invention will now be described in further detail, purely by way of example, with reference to the accompanying drawings, in which:

Fig. 1 schematically show a first embodiment in a longitudinal section,

Fig. 2 schematically section along line II-II in Figure 1 Fig. 3 schematically a second embodiment in longitudinal section,

Fig. 4 a perspective view demonstrate a circular flow pattern 10,

Fig. 5 schematically an embodiment in a longitudinal section,

Fig. 6 schematically a fourth embodiment in a longitudinal section,

Fig. 7 to Fig. 10 schematically a device of the invention wherein closed valves are represented by an unfilled double triangle, and open valves are represented by a filled double triangle to demonstrate the method, Fig. 11 a device of any embodiment of the invention as part of a system of a number of modules.

Description of the Figures

[0031] The figures schematically show a container 1 which may be a module disclosed in WO 2020/098960 Al or especially a process module disclosed in the latter discloser. The container 1 comprises a chamber 2 which is sealed to the environment. The chamber 2 comprises an upper section forming an inlet section 7 and a lower section forming an outlet section 8 or plenum. There may be an intermediate permeable wall 12 through the section of the chamber 2 immediately upstream of the outlet section 8. Inlet conduit 4 is connected with the inlet section 7. The inlet section 7 comprises a first gas flow generator 3 which may comprise one or more fans, nozzles or baffles. Downstream with respect to the inlet conduit 4 for a filter 5 is located. The filter 5 may be an HEPA-filter and may fill the full section of the chamber 2. The filter 5 may act as a diffuser.

[0032] The first gas flow generator 3 generates a laminar gas flow through the chamber 2. The flow pattern is indicated as 9. An axis A is defined by the flow direction of the laminar gas flow.

[0033] A second gas flow generator 6 comprising at least one fan and optionally comprising nozzles or baffles is located inside or outside the chamber 2. The second gas flow generator 6 may be located inside the chamber 2 in that way that the second gas flow generator 6 can deviate or disturb the laminar gas flow (figure 1). The laminar gas flow may have flow velocities less than 0.3 to 0.5 m/s. The second gas flow generator 6 accelerates the gas flow and changes the direction of the gas flow. The second flow generator 6 can generate a gas flow with a flow direction having an angle between 60 and 120° to the first flow direction or the direction of the axis A. [0034] The second gas flow generator 6 is preferably located beside the chamber as shown in figures 3 to 6. The second gas flow generator generates an independent gas flow with respect to the first gas flow. The second gas flow generator may have a fan to accelerate an airflow into the chamber 2.

[0035] The drawings show a second flow generator 6, which produces a gas flow being perpendicular to the axis A. The second flow generator 6 produces a circular flow pattern 10. The second flow generator 6 produces a turbulent flow pattern like a vortex.

[0036] A fumigant generator 14 generates a fumigant, which is a sterilisation gas, preferably hydrogen peroxide. The fumigant is heated by a heater 13 being located downstream of the fumigant generator 14. Pump or fan 17 conveys the fumigant through the inlet conduit 4 into the inlet section 7.

[0037] The fumigant passes through the chamber 2 alternating as a laminar flow or as a turbulent flow. First steps, wherein a laminar flow is provided, change with second steps, wherein a turbulent flow is provided.

[0038] The fumigant leaves the outlet section 8 through an outlet conduit 11. The fumigant may pass through a conduit 19 to the heater 13 and back to the inlet conduit 4 to be fed into the inlet section 7 again.

[0039] After finishing the fumigation the fumigant passes into a scrubber 15, where the fumigant is decomposed. There may be an additional pump or fan 18 to convey the gas. The scrubber 15 may be a catalytic scrubber.

[0040] There may be a sensor 16 sensing residual sterilisation gas in the gas flow exiting the scrubber 15. Additionally, the drawings show several valves for switching a flow path. A controller 20 is provided for controlling the fans of the gas flow generators 3, 6, 18, 31 as well as the heater 13, the fumigant generator 14 and the valves. The controller 20 may have a control program for alternating the flow inside the chamber 2 between a laminar flow and turbulent flow and for heating the fumigant.

[0041] Figure 3 shows a process module of a cell culture system as shown in figure 11. The device shown in figure 3 differs from the device shown in figure 1 by a different arrangement of the second gas flow generator 6 which is a fan generating a gas flow through a wall of the container 1. An opening of the chamber wall forms the nozzle 34, which forms a flow connection between a housing in which the fan 6 is located and the chamber 2. The opening 34 or the fan 6 is located close to a corner of the chamber wall for generating a circular flow around the axis A as shown in figure 4 schematically.

[0042] An outer fan 31 which may be inside a separate housing together with an additional HEPA-filter 32 is connected via a conduit and two valves 23, 24 with the housing containing the fan 6. The outer fan 31 can generate a negative pressure to suck air or fumigant out of the chamber 2.

[0043] A mobile device 30, comprising a fumigant generator 14 and a scrubber 15, can be connected to the pipe system of the module via detachable pipe connections 29. The fumigant generated by the fumigant generator flows via a conduit and through valve 23 into the housing of the fan 6 to generate a turbulent flow of fumigant inside the chamber 2.

[0044] A fan 18 is located downstream of the outlet conduit 11 to accelerate an outlet gas flow via the detachable pipe connection 29 into the scrubber 15 via valve 27. Alternatively, the outlet gas flow can flow through a valve 21 and a valve 22 to the inlet section 7 or through a valve 23 to the chamber comprising the fan 6.

[0045] The outlet conduit 11 has a flow connection to a sensor 16 in which a valve 28 is arranged.

[0046] The arrangement shown in figure 3 comprises a process module, and H2O2 source and H2O2 sink, and a pneumatic circuit required for fumigation. Suitable apparatuses are known for the H2O2 source and sink, such as a product of the company Bioquell.

[0047] An air inlet 37 is connected via a valve 25 with the inlet conduit 4 for feeding air into the chamber 2. The air is distributed by the filter 5 homogeneously into the chamber 2 with a relatively slow velocity of 0.5 m/s or below. The chamber 2 may have an array of appropriated holes. Such an array 12 can be located upstream of the outlet section being formed by a plenum 8. The laminar flow can be established in the chamber 2 which allows efficient gas exchange. The chamber 2 may be further be provided with a duct to recirculate air from the outlet section 8.

[0048] The fan 18 is in line between the outlet section 8 and the fan 3. The fan 18 conveys the gas from a plenum which is formed by the outlet section 8 and which has the same footprint as the chamber 2 above the array and the footprint of the filter 5.

[0049] The device shown in figure 5 differs from the device shown in figure 3 in the way that the fumigant generator 14 and the scrubber 15 are fixedly joined with the device. [0050] The device shown in figure 6 differs from the device shown in figure 3 or 5 by a flow channel 35 distributing the gas flow generated by fan 6 into three flow paths which may be formed by nozzles 34. The gas, in particular air, flowed through the nozzles 34 into the chamber 2 forms turbulent flow pattern 10 as shown in figure 4.

[0051] Laminar flow enables efficient gas exchange, however, during laminar flow, a "boundary layer" of air tends to get trapped on surfaces (e.g. of equipment), and exposure of the surfaces, e.g., to H2O2, or removal of H2O2 from the surfaces, therefore relies on diffusion through the boundary layer, which is relatively slow. Also, while "dead spots" can be minimised in a laminar flow regime, air may be trapped in cavities in and around equipment, and this air will also be exchanged slowly in laminar flow regimes.

[0052] Figure 5, showing air flowing through nozzles 34 or apertures, e.g. spaced down one or two corners, so as to generate a flow that rotates around the central vertical axis A of the process module chamber.

[0053] There is provided a means for generating faster, turbulent, airflow, preferably an orthogonal flow, to mix air out of dead spots, and disrupt boundary layers. In a preferred implementation, there is provided an apparatus to generate a second airflow, which is orthogonal to the laminar air flow, to favor sweeping air from pockets that are dead spots with respect to the laminar flow, and is also faster, so as to favor turbulent mixing. The means may include e.g. a series of holes or nozzles 34 in a duct that is routed up one corner of the chamber 2, arranged to drive an airflow rotationally about the central vertical axis A of the chamber 2. There is further a means for driving airflow, such as a fan or pump. In an alternative implementation, the fans above the HEP A filter may be driven faster, or reversed, or ramped up and down in speed, to generate turbu- lent airflow. In another alternative embodiment, there could be exposed fans in the chamber, rather than ducts and nozzles.

[0054] There is an H2O2 sensor 16 provided, to monitor H2O2 concentration on the outlet, and a valve 28 provided to isolate the sensor 16 from the initial, high concentrations of H2O2, which may be damaging to the sensor.

[0055] In order to increase efficiency and convenience, some means may be provided to direct airflow through the ducts, such as valves 21 to 28, but other means are known in the art, and other pneumatic circuits are possible.

[0056] Other mechanisms, such as heat and UV LEDs may further be provided to further accelerate H2O2 destruction.

[0057] The method is shown in figures 7 to 10. In these figures some of the circuit is omitted for clarity (i.e., the HEP A filtered air outlet, and the air inlet).

Closed valves are represented by an unfilled double triangle, and open valves by a filled double triangle.

[0058] Typically, the system will be connected, e.g. by means of tubing and fittings, such as camlock fittings, to an apparatus that can generate H2O2, and after fumigation is complete, can destroy residual H2O2.

[0059] Figure 7 shows the start of the fumigation. Firstly, H2O2 vapour is injected, by opening valves 26 and 22, and running fans 3 (and optionally fan 18). This results in air from the system being recirculated through the H2O2 generator 14, which injects H2O2 vapour. Initially, it is desired to quickly and efficiently exchange the air in the system, with air that contains H2O2 vapour, so airflow is driven through the HEPA filter, by opening valve 22 and running fans 3. The HEPA filter 5 acts as a diffuser, and if the air flow in the chamber 2 is below about 0.5m/s, then conditions are generally favourable for laminar flow. Injection of H2O2 vapour under laminar flow may be conveniently continued for 5 to 30 seconds, or longer, to allow time for stable laminar flow to develop, and air exchange, but in typical conditions, e.g. at 0.5 m/s, with less than two linear meters height for the air to travel in the chamber 2, the system air should be exchanged in seconds.

[0060] Under laminar flow, as mentioned previously, equipment in the chamber 2 is likely to disturb the laminar flow 9, resulting in dead pockets of air, and usually, there will be a boundary layer of air on surfaces that is not rapidly exchanged with air containing H2O2 vapour. Therefore, during fumigation, it is advantageous to alternate laminar flow with orthogonal turbulent flow 10. The orthogonal turbulent flow 10 (see figure 8) can be generated by closing valve 22 (and stopping fans 3), and opening valve 23 and running fan 6. If valve 26 is open, then H2O2 vapour can still be injected during the orthogonal turbulent flow. As before, fan 23 can optionally be run.

[0061] The conditions required to achieve turbulent flow will depend to some extent on the chamber 2, and on the size and shape of the nozzles or apertures. Flowing air through an aperture or nozzle, rather than across the whole area of a wall, will tend to create a jet, and the shear region between the air jet and surrounding air can generate turbulence, depending of the relative velocities. Importantly, further turbulence will be generated as the moving air passes by obstructions, such as equipment. Constricting the airflow at the nozzle will speed the airflow up, and the shape of the nozzles will also affect how well surrounding air is entrained. Suitable conditions are usually found by injecting smoke into the airflow, and increasing the airspeed until suitable conditions are found. However, airflows above 0.5 to 1 m/s, from nozzles, will typically start to pro- duce turbulent flow, and the effectiveness of the turbulent mixing will increase as the air speed increases. Turbulent mixing will be established in seconds, so turbulent flow may conveniently be run for 5 to 30 seconds or more.

[0062] Therefore, it is convenient to fumigate under alternate laminar and turbulent flow, with laminar flow at less than 0.5 m/s, for 5 to 30 seconds, and turbulent mixing in excess of 0.5 to 1 m/s, for 5 to 30 seconds. This is repeated for the time required for fumigation, e.g. 5 to 10 minutes.

[0063] If the length of time for fumigation exceeds the time during which H2O2 is injected, the air containing H2O2 vapour can be recirculated through the system. For recirculation under laminar flow, valves 21 and 22 are opened, and fans 3 (and optionally Fan 18) are run. For recirculation under turbulent flow, Valves 21 and 23 are opened, and fan 6 (and optionally fan 18) are run.

[0064] As is known in the art, fumigation may further be accelerated by other means, such as heating the vapour.

[0065] Once fumigation is complete, the H2O2 must be purged from the chamber 2, before the next biological process can be run.

[0066] To purge the chamber, (see figure 9) valve 27 is opened to allow recirculation through the catalytic scrubber 15, or some other means to remove the H2O2. Alternatively, the air containing the H2O2 vapour may be exhausted through a vent. To recirculate air from the chamber 2 through the scrubber 15, valve 22 is opened, and fans 3 are run. It may be advantageous to also run fan 18 and close valve 21. As described previously, when fans 3 are run slowly enough for airflow in the chamber to be below ~0.5 m/s, this will usually result in substantially laminar flow, which may leave dead pockets of air, and leave boundary layers of air undisturbed on surfaces.

[0067] Therefore, the laminar flow is alternated with fast, orthogonal turbulent flow, by closing valve 22, turning off fans 3, opening valve 23, and running fan 6. Airflow should be above 1 m/s (see figure 10).

[0068] In this way, purging is achieved by recirculating the air from the system through the catalytic scrubber, while alternating between laminar flow and turbulent, mixing in the chamber with laminar flow for 5 to 30 seconds or more at less than 0.5 m/s, and turbulent mixing flow for 5 to 30 seconds or more at 0.5 to 1 m/s or greater. Multiple cycles of alternating laminar flow and turbulent mixing are run, while recirculating air from the system through the catalytic scrubber 15. Once the H2O2 levels are predicted to be low enough not to harm the H2O2 sensor, valve 28 can be opened to expose the H2O2 sensor to levels of H2O2 vapour coming from the system. This allows the purging to continue until measured H2O2 vapour levels reach, or are below, a pre-defined threshold level.

[0069] As is known in the art, purging may further be accelerated by other means, such as heating the vapour, and exposing the vapour to UV light, e.g. from UV LEDs.

[0070] Figure 11 schematically shows a system comprising several modules of a cell culture system for handling, maintaining or storing biological cells in microplates. A process module 1 may be realised as shown in figures 1 to 5. The processor module contains equipment as mentioned above. Typically, material for a workflow is loaded from storage modules 36 into the process module, the process is run, and material is unloaded from the process module, leaving the process module ready for the next process. [0071] The above explanations serve to explain the inventions covered by the application as a whole, which further develop the state-of-the-art at least through the following combinations of features, each of which may also be dependent, whereby two, several or all of these combinations of features may also be combined, mainly:

[0072] An apparatus, characterised by at least one second gas flow generator 6 for generating a turbulent gas flow in said at least one region.

[0073] An apparatus, characterised in that a heater 13 is provided for heating the fumigant or air prior entering said region.

[0074] An apparatus, characterised by at least one filter 5 being arranged in the flow path of the fumigant or air prior entering said region, wherein the filter 5 is preferably a HEPA filter.

[0075] An apparatus, characterised in that the first gas flow generator 3 produces a gas flow with a first flow direction and the second gas flow generator 6 is located inside said chamber or outside said chamber and produces a gas flow in a second flow direction being different from the first flow direction.

[0076] An apparatus, characterised in that the second flow direction comprises an angle between 60 and 120° to the first flow direction and preferably an angle of 90°.

[0077] An apparatus, characterised in that the second gas flow generator 6 deviates the gas flow produced by the first gas flow generator 3 or generates a gas flow independent from said laminar gas flow. [0078] An apparatus, characterised in that a conduit 11 connects an outlet section 8 of said chamber 2 with a scrubber in particular 15 with a catalytic scrubber 15.

[0079] An apparatus, characterised by a controller 20 for alternately switching between laminar flow condition and turbular flow condition inside said at least one region.

[0080] An apparatus, characterised by a sensor 16 in particular located downstream of a scrubber 15 being sensitive to the fumigant.

[0081] An apparatus, characterised in that the first and/ or second gas flow generator 3, 6 comprises a fan, a nozzle 34 and/ or a baffle.

[0082] An apparatus, characterised in that said chamber 2 is coupled to at least one storage module 36 for storing microplates, cell cultures, lab ware and/ or handling devices.

[0083] An apparatus, characterised in that the inlet section 7 is located in an upper section of the container 1 and/ or that the outlet section 8 is located in a lower section of the container 1.

[0084] An apparatus, characterised in that the second gas flow generator 6 is located outside the chamber 2 and is in a flow connection to the chamber 2 by an opening 34 in a side wall of the chamber 2.

[0085] An apparatus, characterised by a number of openings 34 in a side wall of the chamber 2 being located in different distances from the inlet section 7, wherein each of said openings 34 is in a flow connection with a flow channel 35 for distributing a gas flow generated by the send gas flow generator to the openings 34.

[0086] A method, characterised by switching alternately between said laminar flow and a turbular flow at least inside said region.

[0087] A method, characterised in that the laminar flow is generated by at least one first flow generator 3 and that the turbular flow is generated by at least one second flow generator 6.

[0088] A method, characterised in that the first gas flow generator 3 generates a laminar flow in a first flow direction and the second gas flow generator disturbs the laminar gas flow or generates an independent gas flow in a second flow direction being different from the first flow direction.

[0089] A method, characterised in that the second flow direction comprises an angle between 60 and 120° to the first flow direction and preferably an angle of 90°.

[0090] A method, characterised in that the fumigant comprises a sterilisation gas, in particular H2O2.

[0091] A method, characterised in that the chamber 2 is purged with a purge gas, in particular air, after applying the fumigant to the chamber 2.

[0092] A method, characterised in that during purging the chamber 2 a purge gas flow is altered between a laminar and a turbulent flow. [0093] A method, characterised in that UV-light is used for a degradation of the fumigant after exhausting the fumigant out of the chamber.

[0094] A method, characterised in that a gas flow comprising the fumigant is recirculated through the chamber 2 by using a conduit 19 connecting the outlet section 8 with the inlet section 7.

[0095] A method, characterised in that a first step with a laminar flow has a duration of 5 to 30 seconds and in that a second step immediately following the first step with turbular flow has a duration of 5 to 30 seconds.

[0096] A method, characterised in that sequences of first and second step are repeated for 5 to 10 minutes.

[0097] A method, characterised in that the flow velocity of the laminar gas flow inside the chamber 2 is less than 0.3 to 0.5 m/s and/ or that the second gas flow generator 6 produces flow velocities at least two times higher than the velocity of the laminar gas flow.

[0098] A method, characterised in that sensing of the sterilisation gas downstream of a scrubber 15 by using a sensor 16.

[0099] All disclosed features are (individually, but also in combination with each other) essential to the invention. Hereby, the disclosure of the application also includes the disclosure content of the associated/ attached priority documents (copy of the prior application) in full, also for the purpose of including features of these documents into claims of the present application. The subclaims characterise, even without the feature of a referred claim, with their features independent inventive further developments of the state-of-the-art, in par- ticular in order to make divisional applications on the basis of these claims. The invention indicated in each claim may additionally have one or more of these features indicated in the above description, in particular those provided with the reference numbers and/ or indicated in the list of reference numbers. The invention also concerns forms of design, in which some of the features mentioned in the above description are not realized, in particular insofar as they are recognizably unnecessary for the respective purpose of use or can be replaced by other means having the same technical effect.

Reference Numerals

1 container 27 valve

2 chamber 28 valve

3 first gas flow generator, fan 29 detachable pipe connection

4 inlet conduit 30 desinfection unit, movable

5 filter, HEPA filter 31 outlet fan

6 second gas flow generator, fan 32 outlet filter

7 inlet section 33 air inlet

8 outlet section 34 nozzle, baffle

9 laminar flow pattern 35 flow channel

10 tubular flow pattern, vortex 36 module

11 outlet conduit 37 air inlet

12 intermediate permeable wall

13 heater

14 fumigant generator, sterilisation gas generator A Axis

15 scrubber, catalytic scrubber

16 sensor

17 pump, fan

18 pump, fan

19 conduit

20 controller

21 valve

22 valve

23 valve

24 valve

25 valve

26 valve

Claims

25

Claims

1. Apparatus for handling cell cultures comprising a chamber (2) being arranged in a container (1) and means for providing a flow of a fumigant through at least one region of said chamber (2), wherein said means comprising a fumigant generator (14) being connected with an inlet conduit (4) to an inlet section (7) of said chamber (2), wherein at least one first gas flow generator (3) is provided for generating a laminar flow of said fumigant or of air through said at least one region, characterised by at least one second gas flow generator (6) for generating a turbulent gas flow in said at least one region.

2. Apparatus according to claim 1, characterised in that a heater (13) is provided for heating the fumigant or air prior entering said region.

3. Apparatus according to one of the preceding claims, characterised by at least one filter (5) being arranged in the flow path of the fumigant or air prior entering said region, wherein the filter (5) is preferably a HEPA filter.

4. Apparatus according to one of the preceding claims, characterised in that the first gas flow generator (3) produces a gas flow with a first flow direction and the second gas flow generator (6) is located inside said chamber or outside said chamber and produces a gas flow in a second flow direction being different from the first flow direction.

5. Apparatus according to claim 4, characterised in that the second flow direction comprises an angle between 60 and 120° to the first flow direction and preferably an angle of 90°. Apparatus according to one of the preceding claims, characterised in that the second gas flow generator (6) deviates the gas flow produced by the first gas flow generator (3) or generates a gas flow independent from said laminar gas flow. Apparatus according to one of the preceding claims, characterised in that a conduit (11) connects an outlet section (8) of said chamber (2) with a scrubber in particular (15) with a catalytic scrubber (15). Apparatus according to one of the preceding claims, characterised by a controller (20) for alternately switching between laminar flow condition and turbulent flow condition inside said at least one region. Apparatus according to one of the preceding claims, characterised by a sensor (16) in particular located downstream of a scrubber (15) being sensitive to the fumigant. Apparatus according to one of the preceding claims, characterised in that the first and/ or second gas flow generator (3, 6) comprises a fan, a nozzle (34) and/ or a baffle. Apparatus according to one of the preceding claims, characterised in that said chamber (2) is coupled to at least one storage module (36) for storing microplates, cell cultures, lab ware and/ or handling devices. Apparatus according to one of the preceding claims, characterised in that the inlet section (7) is located in an upper section of the container (1) and/ or that the outlet section (8) is located in a lower section of the container (1).

13. Apparatus according to one of the preceding claims, characterised in that the second gas flow generator (6) is located outside the chamber (2) and is in a flow connection to the chamber (2) by an opening (34) in a side wall of the chamber (2).

14. Apparatus according to one of the preceding claims, characterised by a number of openings (34) in a side wall of the chamber (2) being located in different distances from the inlet section (7), wherein each of said openings (34) is in a flow connection with a flow channel (35) for distributing a gas flow generated by the said gas flow generator to the openings (34).

15. Method for fumigation of at least one region of a chamber (2) located in a container (1) especially of an apparatus according to one of the preceding claims, by feeding a fumigant being provided by a fumigant generator (14) or air into said chamber (2) and generating a laminar flow of said fumigant or air through at least said region, characterised by switching alternately between said laminar flow and a turbulent flow at least inside said region.

16. Method according to claim 15, characterised in that the laminar flow is generated by at least one first flow generator (3) and that the turbular flow is generated by at least one second flow generator (6).

17. Method according to one of the claims 15 or 16, characterised in that the first gas flow generator (3) generates a laminar flow in a first flow direction and the second gas flow generator disturbs the laminar gas flow or generates an independent gas flow in a second flow direction being different from the first flow direction. 28 Method according to one of the claims 15 to 17, characterised in that the second flow direction comprises an angle between 60 and 120° to the first flow direction and preferably an angle of 90°. Method according to one of the claims 15 to 18, characterised in that the fumigant comprises a sterilisation gas, in particular H2O2. Method according to one of the claims 15 to 19, characterised in that the chamber (2) is purged with a purge gas, in particular air, after applying the fumigant to the chamber (2). Method according to claim 20, characterised in that during purging the chamber (2) a purge gas flow is altered between a laminar and a turbulent flow. Method according to one of the claims 15 to 21, characterised in that UV- light is used for a degradation of the fumigant after exhausting the fumigant out of the chamber. Method according to one of the claims 15 to 22, characterised in that a gas flow comprising the fumigant is recirculated through the chamber (2) by using a conduit (19) connecting the outlet section (8) with the inlet section (7). Method according to one of the claims 15 to 23, characterised in that a first step with a laminar flow has a duration of 5 to 30 seconds and in that a second step immediately following the first step with turbulent flow has a duration of 5 to 30 seconds. 29

25. Method according to claim 24, characterised in that sequences of first and second step are repeated for 5 to 10 minutes.

26. Method according to one of the claims 15 to 25, characterised in that the flow velocity of the laminar gas flow inside the chamber (2) is less than 0.3 to 0.5 m/s and/ or that the second gas flow generator (6) produces flow velocities at least two times higher than the velocity of the laminar gas flow.

27. Method according to one of the claims 15 to 26, characterised in sensing of the sterilisation gas downstream of a scrubber (15) by using a sensor (16). 28. Apparatus or method, characterised by one or more of the features of one of the preceding claims.