KR100824357B1 - Miniaturized bioreactor - Google Patents

Abstract

A miniaturized bio-reactor is provided to maintain reaction homeostasis with the optimum and have a non-complicated structure with minimized contamination possibility. A bio-reactor for culturing bio-materials comprises: an interface chip(2) including a culture medium supplying micro-pump(2a), a culture medium extraction micro-pump(2b), a mixture gas flow path, a gas-permeable porous plate(2d), a thermo-sensor(2e) and a pH sensor(2f) and equipped at the lower part of the bio-reactor; and a cultivation tank(1) which is installed at the upper part of the interface chip and includes a culturing chamber(1a) for culturing the bio-materials, an inhalation chamber(1b) for extracting used culture medium, and an outlet(1c) connecting the culturing chamber and the inhalation chamber having a cell isolation filter(1d), wherein the thermo-sensor and the pH sensor are connected to a controller installed at the outside. The bio-reactor further comprises a thermoelectric element(4) receiving a signal from the controller to control the temperature. Further, the porous plate is a porous ceramic plate.

Description

Miniaturized Bioreactor

1 is an overall configuration diagram of a small bioreactor according to the present invention,

2 is a detailed configuration diagram of the culture unit of the configuration of Figure 1,

3 is an exploded view of one embodiment of a small bioreactor culture unit according to the present invention.

(Explanation of symbols for the main parts of the drawing)

10: culture unit

1: culture tank 1a: culture chamber

1b: suction chamber 1c: outlet

1d: cell separation filter 2: bioreactor interface chip

2a: Medium supply micro pump 2b: Medium extraction micro pump

2c: mixed gas flow path 2d: gas permeable porous plate

2e: temperature sensor 2f: pH sensor

3: heat transfer plate 4: thermoelectric element

5: heat sink 6: cover

7: push button

20: controller

30: gas mixer

40: filter

50: gas heat exchanger

60: mixed gas reservoir

70: discharge tank

80: wastewater tank

90 filter

The present invention relates to a bioreactor for small-scale cultivation of various biological materials such as various cells, enzymes, proteins, including animal cells, and an interface chip constituting a part thereof. More specifically, the present invention relates to an interface chip constituting the lower portion of the culture tank in a small bioreactor and a bioreactor including the same, the interface chip is a medium supply micro-pump for supplying the medium to the culture tank, the medium used from the culture tank A medium extraction micropump to extract, a mixed gas flow path providing a path of the mixed gas supplied to the culture tank, and a gas permeable porous plate, a temperature sensor, and a pH sensor provided on the mixed gas flow path to supply gas into the culture tank. Include.

Bioreactors are reaction systems or devices that perform biochemical reactions using bioreactors. Especially, in small bioreactors, they are small in volume, which means that they do not respond to rapid changes in the environment, such as temperature, pH, or composition. Since the ability to absorb subsequent shocks is weak compared to large incubators, the presence or absence of a control system that maintains homeostasis in the system depends on the success or failure of the desired bioreaction.

In general, small-scale bioreactions, especially in cell culturing, have a significant effect on the growth of cells, including the concentration of gas in the medium, such as oxygen, carbon, and nitrogen, the pH of the medium, the temperature of the medium, and the supply of fresh medium. The removal of waste products such as spent media, and how to finely control these elements is a challenge for all bioreactors as well as small bioreactors.

In these bioreactions, the elements that require control are primarily sensors that monitor the state of each element in the culture, such as temperature sensors, pH sensors, CO ₂ sensors, dissolved oxygen measurement sensors, etc. When installed and the signal is transmitted to the controller, the controller with on / off and purge control functions according to the difference between the measured value and the reference value to transfer or cool the heat close to the reference value, or to adjust the pH control liquid, oxygen or carbon dioxide. It is carried out in such a way that to deliver into the incubator.

Bioreactors are designed to meet optimal conditions for cultivating biomaterials, so growth of other organisms or contamination of other materials other than the desired biomaterial is always a problem. In particular, bioreactors have various components and structures. The potential for contamination is increased if the product contains parts that are complicated or not easy to clean. In the conventionally used bioreactors, the problem of contamination was often serious due to the repeated reuse of various sensors installed separately and the culture tank itself. Once pollution occurs, not only does the bioreaction itself operating fail, but also the enormous cost and effort required to remove it afterwards. In particular, the cultivation of antibodies or expensive cells in the pharmaceutical field can be enormous.

Therefore, there is an urgent need for a bioreactor having a control system capable of maintaining homeostasis close to a living body while having a simple structure in which pollutants can be blocked as much as possible.

The present invention has been made to solve the problems of the prior art as described above, the object of the present invention is to provide a control system that can optimally maintain the reaction homeostasis of small scale bioreactions, while not structurally complex and minimize the possibility of contamination The present invention provides a small bioreactor having a structure and an interface chip used therein.

The inventors of the present invention have made an amazing discovery that the use of an interface chip manufactured by a MEMS (MicroElectroMechanical Systems) production process, which means a micro system or a micro machine, is very advantageous for increasing the efficiency of a small bioreactor and maintaining the stability of the reaction. Came to complete.

In order to achieve the above object, a small bioreactor according to the present invention includes a bioreactor for culturing a biomaterial, the interface chip provided under the bioreactor; And a culture tank installed at an upper portion of the interface chip and configured to connect a culture chamber for culturing a biological material, an intake chamber for extracting a used medium, and a culture chamber and an intake chamber, and an outlet having a filter. And the temperature sensor and the pH sensor are configured as a bioreactor, characterized in that connected to an externally installed controller.

In addition, the interface chip is an interface chip constituting the lower part of the culture tank in a small bioreactor, a medium supply micropump for supplying a medium to the culture tank, a medium extraction micropump for extracting the medium used from the culture tank, the culture A mixed gas flow path providing a path of the mixed gas supplied to the tank, a gas permeable porous plate provided on the mixed gas flow path to supply gas into the culture tank, a temperature sensor for measuring a temperature inside the culture tank, and Characterized in that it comprises a pH sensor for measuring the pH inside the culture tank.

In one embodiment of the invention the porous plate is characterized in that the porous ceramic plate.

In another aspect of the present invention, the lower portion of the interface chip is characterized in that it further comprises a thermoelectric element capable of temperature control by receiving a signal from the external controller.

In another embodiment of the present invention, the gas supplied to the mixed gas flow path is supplied by mixing at least two gases selected from oxygen, nitrogen, ammonia, and carbon dioxide from a gas mixer installed outside, between the gas mixer and the mixed gas flow path. The gas heat exchanger and the mixed gas reservoir connected to the controller is installed is characterized in that the temperature of the mixed gas can be adjusted.

In another aspect of the present invention, a filter for disinfection is provided between the gas mixer and the mixed gas flow path.

In another aspect of the invention, the culture chamber is characterized in that the baffle is provided.

Hereinafter, with reference to the accompanying drawings, an embodiment of the interface chip constituting the lower part of the culture tank in the small bioreactor and the bioreactor of the present invention will be described in more detail.

(Structure of Bioreactor and Interface Chip)

1 is an overall configuration diagram of a small bioreactor according to the present invention, Figure 2 is a detailed configuration diagram of the culture unit of the configuration of Figure 1, Figure 3 is an exploded view of an embodiment of a small bioreactor culture unit according to the present invention .

As shown in Figures 1 to 3, the small bioreactor culture unit 10 according to the present invention is a culture tank (1) constituting the upper portion of the bioreactor, the interface chip (2) constituting the lower portion, the lower portion In the heat transfer plate 3, the thermoelectric element 4 and the heat dissipation plate 5 for adjusting the temperature of the culture tank. The thermoelectric element 4 serves to maintain the culture vessel and the culture vessel component parts at a uniform temperature through the heat transfer plate 3.

The interface chip (2) constituting the lower part of the culture tank is a medium supply micropump (2a) for supplying the medium to the culture tank (1), a medium extraction micropump (2b) for extracting the medium used from the culture tank , A mixed gas flow path 2c providing a path of the mixed gas supplied to the culture tank 2, a gas permeable porous plate 2d provided on the mixed gas flow path to supply gas into the culture chamber, and It consists of a temperature sensor 2e for measuring the temperature inside the culture chamber 2a and a pH sensor 2f for measuring the pH inside the culture chamber 2a.

The bioreactor of the present invention including the interface chip 2 is installed on the interface chip 2 provided on the lower portion of the bioreactor, and the interface chip, and a culture chamber 1a for culturing a biomaterial. ), A culture tank 1 composed of an inlet chamber 1b for extracting the used medium and an outlet port 1c having a cell separation filter 1d, which serves to connect the culture chamber and the suction chamber.

The temperature sensor 2e and the pH sensor 2f are configured to be connected to the controller 20 installed outside.

The medium supply micropump 2a is configured to pump from the medium tank 70 into the culture chamber 1a, and the medium extraction micropump 2b is configured to pump from the suction chamber 1b to the wastewater tank 80. do.

The gas supplied to the mixed gas flow path 2c is supplied by mixing two or more kinds of gases selected from oxygen, nitrogen, ammonia and carbon dioxide from the gas mixer 30 installed outside, and the gas mixer 30 and the mixed gas. The gas heat exchanger 50 and the mixed gas reservoir 60 connected to the controller 20 are installed between the flow paths 2c.

A filter 40 for disinfection is installed between the gas mixer 30 and the mixed gas flow path 2c.

A filter 90 for disinfection may be installed at the outlet of the mixed gas flow path.

The operation of the small bioreactor and the interface chip according to the present invention having such a configuration will be described in more detail with reference to the accompanying drawings.

(Operation of small bioreactors)

In the bioreactor of the present invention as shown in FIG. 1, various biomaterials including various cells, enzymes, antibodies and other proteins in the culture chamber 1a are cultured on a medium supplied from the medium tank 70. It is suitable for small scale cultures of less than 100ml. The medium used in the culture chamber is supplied by a medium supply micro pump 2a on the interface chip 2, which pumps the medium from the medium tank.

The culture chamber (1a) is connected to the suction chamber (1b) and the outlet (1c) for the extraction of the used medium, the outlet to prevent the culture material, such as cells to flow out when the discharge of the used medium To this end, a cell separation filter 1d having a pore size through which the culture material cannot pass, preferably having a pore size of about 0.2 μm, is installed so that the medium used through the filter is transferred to the wastewater tank 80 through the suction chamber. Is moved. At this time, the power for discharging is provided by the medium extraction micropump 2b installed in the interface chip.

The mixed gas is supplied into the culture tank 2 from the mixed gas flow path 2c, which is provided on the mixed gas flow path through a gas permeable porous plate 2d for supplying gas into the culture tank. The mixed gas is determined from the type and amount of gas required by the controller 20 installed outside the culture unit, and is supplied from the gas mixer 30. The gas permeable porous plate may preferably be a porous ceramic plate.

On the other hand, the waste gas from the medium in the culture chamber diffuses into the outside air through the gas permeable porous plate 2d. Unused mixed gas and waste gas are discharged to the atmosphere through a gas outlet (not shown) of the interface chip 2.

Here, the controller processes the temperature information and the pH information from the temperature sensor 2e and the pH sensor 2f installed facing the culture tank on the interface chip to control the temperature with the thermoelectric element 4, or the gas mixer. At 30, it is operated to supply a required amount of gas such as CO ₂ or NH ₃ for pH adjustment.

Cultivation conditions of the culture material are measured not only with the sensor but also with a dissolved oxygen measurement sensor (not shown) installed so that the amount of O _{2 that} must be supplied from the gas mixer is also driven through the controller.

In addition, the gas mixer 30 supplies the respiratory gas, such as O ₂ , N ₂ , into the culture tank through the mixed gas flow path 2c according to the control of the controller. At this time, the mixed gas flow path is configured in a labyrinth, it is possible to more efficiently supply the mixed gas into the culture chamber (1a).

In the case of the small-volume culture tank, the temperature of the culture tank is drastically changed by the gas introduced into the culture tank, so that the supply gas is also supplied under the control of the controller 20 by controlling the temperature with the heat exchanger 50 using the thermoelectric element. In particular, a reservoir tank 60 is installed at the end of the heat exchanger to equalize the heat flow of the feed gas.

On the other hand, the upper part of the culture tank (1) can check the culture state with the naked eye and a cover 6 for sealing the culture tank is installed, the transparent cover, preferably a glass cover to visually check the state in the culture tank Can be installed. In order to seal the culture tank by the cover, a pair of presses 7 for assembling the culture tank 1, the interface chip 2, and the lower heat transfer plate 3 may be installed.

The lower part of the heat transfer plate 3 is assembled with a thermoelectric element, a heat sink and a heat radiating fan (not shown). Here, the cover and the culture tank can be manufactured for single use, and the interface chip produced by the MEMS (MicroElectroMechanical Systems) production process, which means a micro system or a micro machine, can be reused.

A baffle (not shown in the figure) may be provided in the culture chamber 1a, and the culture liquid gradually moves to the suction chamber 1b side through the baffles. The baffle can take the place of agitation of the culture in small reactors by increasing the path of the medium and prevents local retention of fluid in the culture chamber.

The small fermenter according to the present invention having the configuration as described above is a medium supply micropump and a medium extraction micropump closely controlled by a controller, a gas permeable porous plate and a temperature and pH sensor provided on the mixed gas flow path and the flow path. By the action of the interface chip according to the present invention including a fast temperature and gas control and supply is possible, there is an effect that the problems of the prior art, such as stability and contamination of the bioreactor is improved.

In addition, the structure in which a small gas reservoir whose temperature is controlled by the thermoelectric element is installed before the gas supply is directly supplied from the gas mixer has an effect of avoiding a sudden temperature change affecting the bioreaction.

Claims (8)

In the interface chip constituting the lower part of the culture tank in a small bioreactor, A medium supply micropump for supplying a medium to the culture tank; A medium extraction micropump for extracting the used medium from the culture tank; A mixed gas flow path providing a path of the mixed gas supplied to the culture tank; A gas permeable porous plate provided on the mixed gas flow path for supplying gas into the culture tank; A temperature sensor for measuring a temperature in the culture tank; And PH sensor for measuring the pH of the culture tank; Interface chip of the bioreactor comprising a. The interface chip of a bioreactor according to claim 1, wherein the porous plate is a porous ceramic plate. In the bioreactor for the cultivation of biological material, An interface chip of claim 1 or 2 provided below the bioreactor; And The culture tank is installed on top of the interface chip, the culture chamber for culturing the biological material, the suction chamber for the extraction of the used medium and the connection between the culture chamber and the suction chamber, consisting of an outlet having a cell separation filter The bioreactor comprising a, wherein the temperature sensor and the pH sensor is connected to an externally installed controller. The bioreactor of claim 3, further comprising a thermoelectric element at a lower portion of the interface chip that is capable of temperature control by receiving a signal from the external controller. The gas supplied to the mixed gas flow path is supplied by mixing at least two kinds of gases selected from oxygen, nitrogen, ammonia, and carbon dioxide from a gas mixer installed outside, between the gas mixer and the mixed gas flow path. Bioreactor characterized in that the gas heat exchanger (50) and the mixed gas reservoir (60) connected to the controller is installed to control the temperature of the mixed gas. 4. The bioreactor of claim 3, wherein a filter for disinfection is provided between the gas mixer and the mixed gas flow path. The bioreactor of claim 3, wherein the culture chamber is provided with a baffle. The bioreactor of claim 3, further comprising a cover that is visible at the upper portion of the culture tank, wherein the culture tank and the cover are made disposable.

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