WO2017126237A1

WO2017126237A1 - Device and method for treating culture product and device for purifying culture product

Info

Publication number: WO2017126237A1
Application number: PCT/JP2016/085791
Authority: WO
Inventors: 勝難波; 近藤　健之
Original assignee: 株式会社日立製作所
Priority date: 2016-01-22
Filing date: 2016-12-01
Publication date: 2017-07-27
Also published as: JP2017127283A

Abstract

Provided are a device for treating a culture product and a method for treating a culture product, whereby a virus inactivation treatment can be efficiently carried out as a substitute for a conventional low pH treatment. Also provided is a purification device for a culture product that comprises the device for treating a culture product. The device (3) according to the present invention for treating a culture product comprises: a flat channel (5) provided with an inlet port (5b) and an outlet port (5c) through which a liquid culture medium, said liquid culture medium being free from cells and containing the culture product, flows; a heating part (6) which is disposed in contact with or close to one surface of the channel (5) and which heats the liquid culture medium; and a ultraviolet irradiation part (8) which is disposed at a position facing the heating part (6) in contact with or close to the other surface of the channel (5) and which irradiates ultraviolet rays to the liquid culture medium.

Description

Cultured product processing apparatus, cultured product processing method, and cultured product purification apparatus

The present invention relates to a culture product processing apparatus, a culture product processing method, and a culture product purification apparatus.

For example, in the manufacturing process of biopharmaceuticals, beverages, and foods, a culture method for culturing living cells such as microorganisms, cells, and fungi is used. In the case of biopharmaceuticals, there is a risk of contamination of viruses derived from the medium components used for culturing living cells in addition to viruses inherent in living cells used for culturing and viruses due to accidental contamination in the manufacturing process. In the purification process of recovering culture products such as antibody proteins from the culture medium in which cells are cultured, sterilization that reliably reduces bacteria that may be mixed in the culture liquid in order to ensure the safety of the culture product There is a need for inactivation treatments that reliably reduce processing and viruses. Here, the virus inactivation process means a process for losing the infectivity of the virus.

Conventionally, an affinity chromatograph in which protein A, which is an Fc receptor derived from a microorganism, is immobilized as an affinity ligand is generally used for antibody purification. In antibody purification, the antibody is adsorbed to protein A under neutral conditions, and non-adsorbed components are washed and removed, and then the adsorbed antibody is eluted from the ligand and collected under acidic conditions of pH 3 or lower. Such low pH conditions can also be used for virus inactivation treatment. Therefore, in general, by maintaining an acidic eluent at low pH conditions, virus inactivation treatment (referred to herein as “low pH treatment”). ").

However, it is known that when a low pH treatment is performed, the higher-order structure of the antibody changes, the antibody is inactivated or aggregates are formed, and the quality of the antibody may be reduced. In recent years, in order to avoid antibody inactivation and aggregate formation due to low pH treatment, the development of functional carriers such as temperature-responsive affinity chromatographs with different adsorption / desorption characteristics depending on temperature has been promoted. When such a functional carrier is used, a virus inactivation treatment is required in place of the low pH treatment.

As such virus inactivation treatment, for example, the invention described in Patent Document 1 has been proposed. This Patent Document 1 describes a continuous method of sterilizing a fluid medium and optionally inactivating viruses by applying a combination of heat treatment and UV irradiation treatment. Further, Patent Document 1 describes that in a continuous method, heat treatment of a fluid medium is performed at a sterilization temperature of 40 to 135 ° C., and irradiation treatment is performed at an irradiation density of 5 to 300 W / m ² .

Further, this Patent Document 1 discloses a deformable, helical profile in which each of the heat treatment reactor and the UV irradiation reactor is tightly drawn into the wall of a hard, straight cylindrical support. It is described that it is made from a hollow cylinder having Patent Document 1 discloses that each of the heat treatment reactor and the UV irradiation reactor includes a sterilization and / or inactivation chamber through which a fluid medium passes, and a cylindrical support of the heat treatment reactor Made from a thermally conductive material, the cylindrical support of the UV irradiation reactor is stated to be transparent to UV irradiation.

JP 2004-321784 A

However, in the invention (for example, the apparatus) described in Patent Document 1, since each of the heat treatment reactor and the UV irradiation reactor is provided, these treatments cannot be performed simultaneously, and the virus inactivation treatment is efficiently performed. There was a risk that it could not be done.

The present invention has been made in view of the above situation, and a culture product treatment apparatus, a culture product treatment method, and culture production capable of efficiently performing virus inactivation treatment instead of conventional low pH treatment. It is an object of the present invention to provide a product purification apparatus.

An apparatus for treating a cultured product according to the present invention that has solved the above problems is a flat flow path having an inlet and an outlet through which a culture solution containing the cultured product is passed, from which cells have been removed. And a heating part that heats the culture solution and a position facing the heating part on the other surface of the flow path part. An ultraviolet irradiation unit that is provided in contact with or close to the medium and that irradiates the culture solution with ultraviolet rays.

Further, the culture product processing method according to the present invention includes a flat flow path portion provided with an inlet and an outlet through which a culture solution containing the culture product is passed, and from which cells have been removed. It is provided in contact with or in proximity to one surface of the flow channel, and in contact with or in proximity to the heating unit that heats the culture medium and the other surface of the flow channel that faces the heating unit An ultraviolet irradiation unit that irradiates the culture solution with ultraviolet rays, and the culture product processing apparatus using the culture product processing apparatus comprising: While being heated by the heating section, the ultraviolet light is irradiated from the ultraviolet irradiation section to the culture solution in the flow path section.

Furthermore, the culture product purification apparatus according to the present invention is an apparatus for purifying a culture product produced by culturing cells, wherein the cells are removed and a culture solution containing the culture product is passed through. A flat flow path portion having an inlet and an outlet, and a heating section that is provided in contact with or close to one surface of the flow path section, heating the culture solution, and An apparatus for treating a cultured product, comprising: an ultraviolet irradiation unit that irradiates ultraviolet rays together with heating of the culture solution by the heating unit, provided on the other surface in contact with or close to a position facing the heating unit And a chromatograph provided at least before and after the culture product treatment apparatus on the basis of the flow direction of the culture solution.

The culture product treatment apparatus, culture product treatment method, and culture product purification apparatus according to the present invention can efficiently perform virus inactivation treatment instead of conventional low pH treatment.

It is the schematic explaining the whole structure of the refiner | purifier which concerns on one Embodiment of this invention. It is the schematic which shows the example of formation of a flow-path guide. It is the schematic which shows the example of formation of a flow-path guide. It is the schematic which shows the modification of the processing apparatus which concerns on one Embodiment of this invention. It is the schematic which shows the modification of the processing apparatus which concerns on one Embodiment of this invention. It is the schematic which shows the other modification of the processing apparatus which concerns on one Embodiment of this invention.

DETAILED DESCRIPTION OF EMBODIMENTS Hereinafter, a culture product processing apparatus, a culture product processing method, and a culture product purification apparatus according to the present invention will be described in detail with reference to the drawings as appropriate.

[Purification equipment for cultured products]
For convenience of explanation, first, a culture product purification apparatus (sometimes simply referred to as “purification apparatus” in this specification) according to an embodiment of the present invention will be described. FIG. 1 is a schematic diagram illustrating the overall configuration of a purification apparatus 10 according to an embodiment of the present invention.

A purification apparatus 10 shown in FIG. 1 is an apparatus for purifying a culture product produced by culturing cells. This purification apparatus 10 is applied when purifying a culture product obtained by culturing microorganisms or animal and plant cells that produce substances used as main raw materials such as pharmaceuticals and health foods (beverages and foods). be able to.

Examples of cells to be cultured include animal cells, plant cells, photosynthetic bacteria, microalgae, cyanobacteria, insect cells, bacteria, yeasts, fungi, and algae. In particular, animal cells that produce proteins such as antibodies and enzymes are preferably cultured. The medium for culturing the cells is not particularly limited, and an appropriate one can be used according to the cells to be cultured, but a liquid medium is preferable.

Examples of the culture product to be a substance to be produced include, but are not limited to, proteins such as antibodies and enzymes, physiologically active substances such as low molecular compounds and high molecular compounds, and the like. In addition, carotenoids such as β-carotene and astaxanthin, pigments such as chlorophyll and bacteriochlorophyll, phycobilin proteins such as phycocyanin used for coloring foods and cosmetics, and other physiologically active substances such as fatty acids are also cultures to be produced. It can be mentioned as a product.

As shown in FIG. 1, a purification apparatus 10 according to the present embodiment includes a culture product processing apparatus (hereinafter also simply referred to as “processing apparatus”) 3 according to the present embodiment, which will be described later. And chromatograph 9. Specifically, as shown in FIG. 1, the purification apparatus 10 has a culture tank 1, a first storage tank 2a, a processing apparatus 3, a chromatograph 9, and a second storage tank 2b. is doing. These components are arranged in this order, and each component is connected by transfer piping tubes T1, T2, T3, and T4. That is, the culture tank 1 and the 1st storage tank 2a are connected by the transfer piping tube T1, and the 1st storage tank 2a and the processing apparatus 3 are connected by the transfer piping tube T2. Moreover, the processing apparatus 3 and the chromatograph 9 are connected by the transfer piping tube T3, and the chromatograph 9 and the 2nd storage tank 2b are connected by the transfer piping tube T4. Any transfer piping tubes T1, T2, T3, and T4 can be used as long as they can be sent aseptically without contaminating the culture solution. Examples of the chromatograph 9 used in the present invention include a temperature-responsive affinity chromatograph, a high-performance liquid chromatograph (High-Performance-Liquid-Chromatograph; HPLC), and an ion exchange chromatograph.

In addition, in FIG. 1, although the chromatograph 9 which wash | cleans a culture solution has illustrated a mode that it is provided after the processing apparatus 3 (between the processing apparatus 3 and the 2nd storage tank 2b), The arrangement position of the chromatograph 9 is not limited to this. The chromatograph 9 can be provided at least before or after the treatment apparatus 3 in the purification apparatus 10 with reference to the flow direction of the culture solution flowing through the purification apparatus 10. For example, it can be arranged between the culture tank 1 and the first storage tank 2a, that is, on the transfer pipe tube T1, and between the first storage tank 2a and the processing apparatus 3, that is, on the transfer pipe tube T2. Can be arranged. It goes without saying that the chromatograph 9 can be provided both before and after the processing apparatus 3 as required. That is, as shown in FIG. 1, the chromatograph 9 is disposed between the processing device 3 and the second storage tank 2b, and the chromatograph 9 is disposed on the transfer piping tube T1 and / or the transfer piping tube T2. can do. When the culture product to be produced is an antibody and the temperature-responsive affinity chromatograph 9 is employed as the chromatograph 9, it is preferably provided in front of the processing apparatus 3. In this way, the culture solution can be washed in front of the treatment apparatus 3 by the temperature-responsive affinity chromatograph 9. Therefore, it is possible to remove in advance from the culture medium substances that absorb ultraviolet rays, such as proteins and amino acids contained in some viruses, culture solutions, and the irradiation efficiency of the ultraviolet irradiation unit 8 can be improved (ultraviolet and The ultraviolet irradiation unit 8 will be described later).

The culture tank 1 is a container for culturing cells to be cultured as described above and producing a culture product. The culture tank 1 is cultured in a state where optimal temperature and agitation are maintained using a medium whose composition is prepared in advance for culturing target cells. Although not shown in FIG. 1, gas supply equipment such as air, oxygen, nitrogen and carbon dioxide, hot water / cold water supply equipment and water supply / drainage equipment, which are indispensable for the culture tank 1, and in addition to the fed-batch culture Is equipped with a supply facility for an additional medium to be added during culture.

The 1st storage tank 2a is a container which stores temporarily the culture solution cultured by the culture tank 1. FIG. The 2nd storage tank 2b is a container which stores temporarily the culture solution processed with the processing apparatus 3. FIG. The first storage tank 2a and the second storage tank 2b can be made of stainless steel, glass, or the like, but are sterilized in advance by gamma rays, ultraviolet rays, ethylene oxide gas, etc., and maintained in a sterile state, such as gas or liquid. It is also possible to use a commercially available single-use bag that is provided in a clean state with its contents almost removed.
As will be described later in detail, the processing device 3 is a device that performs virus inactivation processing.

Moreover, the purification apparatus 10 according to the present embodiment includes a cell crushing unit that crushes cells as necessary, a filtration filter that removes unnecessary materials such as cells, cell walls of crushed cells, cell membranes, proteins, and a continuous centrifuge. Can be provided (both not shown). These components are preferably provided in front of the processing apparatus 3 in the order of installation. In this way, since the culture solution can be clarified, as described later, it becomes possible to efficiently irradiate the treatment apparatus 3 with ultraviolet rays. That is, it is possible to avoid a situation in which ultraviolet rays are not sufficiently irradiated due to the turbidity of the culture solution. In addition, it is more preferable to provide these components on the transfer piping tube T1. If it does in this way, the inside of the 1st storage tank 2a can be kept clean.

[Processing equipment for cultured products]
Next, an aspect of the processing apparatus 3 according to this embodiment will be described. As already described, the processing apparatus 3 is provided in the purification apparatus 10 described above.
As shown in FIG. 1, the processing apparatus 3 includes a flow path unit 5, a heating unit 6, and an ultraviolet irradiation unit 8. The culture solution flowing through the flow path portion 5 is a cell from which cells have been removed and contains a culture product.

The flow path part 5 is formed so as to be covered by the housing part 4a and the housing part 4b. Moreover, the flow path part 5 has comprised the flat shape provided with the inflow port 5b and the outflow port 5c which flow a culture solution. Here, in this specification, the flat shape means that the dimension in the plane direction (longitudinal dimension and lateral dimension) is long, and the height direction (dimension in the thickness direction between the housing part 4a and the housing part 4b). Is a short shape. Although the dimension of the flow path part 5 in the planar direction is not limited, the dimension in the thickness direction depends on the heating of the culture solution in the flow path part 5 by the heating part 6 described later and the culture in the flow path part 5 by the ultraviolet irradiation part 8. It is preferable to set so that the liquid is appropriately irradiated with ultraviolet rays. The dimension of the flow path portion 5 in the thickness direction is preferably 0.5 to 10 cm, for example, and more preferably 1 to 5 cm.

The heating unit 6 is provided in contact with or close to one surface of the flow channel unit 5 and heats the culture solution in the flow channel unit 5. The heating unit 6 can be composed of an electric heater or the like. The heating temperature of the culture solution by the heating unit 6 is preferably set to a temperature that does not cause thermal denaturation of the culture product. The temperature that does not cause the heat denaturation of the cultured product varies depending on the cultured product, and can be appropriately set according to the cultured product. For example, it can be set to 80 ° C. or the like, but may be set to 75 ° C. or 65 ° C. You can also. When the upper limit of the heating temperature of the culture solution by the heating unit 6 is set as described above, for example, even when the culture product is an antibody protein, heat denaturation is performed while inactivating the virus in the culture solution. It is possible to avoid such a situation that the activity is impaired by. In addition, it is preferable that the minimum of the heating temperature of the culture solution by the heating part 6 shall be 60 degreeC, for example.

Here, with regard to viruses to be subjected to inactivation treatment, a literature (Nims) that investigated heat inactivation characteristics of 12 viruses belonging to non-enveloped viruses that are considered to be relatively resistant to sterilization treatment such as heating. , RW and Plavsic, M .: Intra-family and inter-family comparisons for viral susceptibility to hest inactivation: J. Microb.Biochem.Technol., 5,136-141 (2013)). In this document, viruses exhibiting an inactivation effect with an LRV (logarithmic reduction index) of 4 or more after heating at 70 ° C. for 1 minute are: Caliciviridae (Feline calicivirus, Murine norovirus), Picornaviridae (Foot and mouth disease virus) (strain OPN), Hepatitis A virus, Coxsackie B-5 (Faulker strain)), Birnaviridae (Infectious pancreatic necrosis virus, Infectious bursal disease virus) have been reported. On the other hand, in this document, viruses of the family Parvoviridae (Mouse minute virus, Bovine parvovirus, Canine parvovirus) require a heating time of 40 minutes or more at 70 ° C to obtain a virus inactivating effect of LRV = 4 or more. It has been reported. In order to obtain a virus inactivating effect with a heating time of 1 minute or less and an LRV of 4 or more, a heating temperature of about 110 ° C. is required. Therefore, the processing apparatus 3 according to the present embodiment can set the heating temperature and the heating time in consideration of such information in the virus inactivation process. Here, it cannot be overemphasized that according to the processing apparatus 3 which concerns on this embodiment, disinfection and a sterilization process can be performed simultaneously with the inactivation process of the virus in a culture solution.

In addition, it is preferable that a cooling unit 7 for cooling the culture solution is provided downstream of the heating unit 6 on the basis of the flow direction of the culture solution. The cooling temperature of the culture solution by the cooling unit 7 can be arbitrarily set according to the thermal stability of the culture product and the processing in the subsequent process. The cooling unit 7 can be formed by appropriate means. For example, the cooling section 7 is connected to the cooling block that contacts the flow path section 5 and absorbs the heat of the culture solution, a tube that is connected to the cooling block and distributes a cooling medium such as water, and the tube. A pump that circulates the cooling medium, a radiator that is connected to the tube and cools the cooling medium, and a fan that sends air to the radiator can be used. In addition, for example, the cooling unit 7 may be configured by simply providing a fan that generates an airflow. In this case, since the configuration is simple, the cost can be reduced.

The ultraviolet irradiation unit 8 is provided on the other surface of the flow channel unit 5 so as to be in contact with or close to a position facing the heating unit 6 and irradiates the culture solution in the flow channel unit 5 with ultraviolet light. In this invention, it becomes possible to perform a virus inactivation process effectively by setting it as such a structure. As shown in FIG. 1, the ultraviolet irradiation unit 8 includes an ultraviolet ray source 8a that emits ultraviolet rays (ultraviolet light), an ultraviolet ray reflection unit 8b that reflects the ultraviolet ray emitted from the ultraviolet ray source 8a and efficiently irradiates the culture solution, However, the ultraviolet reflecting portion 8b may not be provided. The wavelength of ultraviolet rays is preferably 254 nm, which is most effective for inactivating viruses, but ultraviolet rays having shorter wavelengths and longer wavelengths are used as long as they are classified as so-called ultraviolet rays. be able to. As the ultraviolet source 8a, for example, a low-pressure mercury lamp, an ultraviolet LED (Light Emitting Diode), or the like can be used. UV dose required for inactivation depends viral species, but generally in the range of 50 ~ 1000J / ^{m 2,} as a dose which does not adversely affect the protein such as an antibody, and 100 ~ 500J / ^{m 2} It is preferable to set the irradiation light intensity and irradiation time of the light source. For example, when an ultraviolet irradiation light source having an ultraviolet irradiation light intensity of 10 W / m ² is used, it is preferable to set the flow rate of the culture solution so that the irradiation time is 10 to 50 seconds. If the ultraviolet irradiation intensity is set within this range, the virus inactivation treatment can be performed more reliably.

And the processing apparatus 3 of the said structure performs the irradiation of the ultraviolet-ray to the culture solution by the ultraviolet irradiation part 8 simultaneously with the heating of the culture solution by the heating part 6. FIG. Therefore, the processing apparatus 3 can efficiently inactivate the virus in the culture solution flowing through the flow path section 5. That is, according to this processing apparatus 3, the virus inactivation process can be efficiently performed in place of the conventional low pH process.

In the present invention, since the culture solution is heated by the heating part 6, the housing part 4b in contact with or in proximity to the heating part 6 is preferably formed using a metal or quartz glass having good thermal conductivity. In addition, since the ultraviolet irradiating unit 8 irradiates the culture solution with ultraviolet rays, the housing part 4a that is in contact with or close to the ultraviolet irradiating unit 8 is formed using quartz glass, resin, or the like that is permeable to ultraviolet rays. preferable.

Further, the processing time by the processing device 3 can be arbitrarily set within a range in which the virus can be inactivated. That is, the processing time by the processing apparatus 3 can be appropriately set according to conditions such as the heating temperature, the ultraviolet irradiation time, and the irradiation intensity. In particular, it is preferable to set the heating temperature, the ultraviolet irradiation intensity, and the treatment time after evaluating the characteristics of virus inactivation for virus species that are expected to be mixed in the cell culture environment. Although not particularly limited, for example, the retention time of the culture solution from the inflow port 5b to the outflow port 5c of the flow path unit 5 is set to 30 seconds or more and 30 minutes or less as a guide for the processing time by the processing apparatus 3. Is mentioned. In this way, reliable virus inactivation processing can be performed. In addition, from the viewpoint of more reliable virus inactivation treatment and efficient treatment, the residence time is preferably 1 minute or more and 20 minutes or less.

Here, FIG. 2A and FIG. 2B are schematic views showing an example of forming the flow path guide 5a. As shown in FIG. 2A, one or more flow path guides 5a can be provided so as to be parallel to the flow direction so as to conform to the flow direction of the culture solution. In addition, as shown in FIG. 2B, one or more flow path guides 5a can be provided so as to be in a direction perpendicular to the flow direction so that the flow of the culture solution meanders. Thus, by providing the flow path guide 5 with the flow path guide 5a that guides the flow direction of the culture solution, the processing time by the processing apparatus 3 can be adjusted. Note that the attachment length and direction of the flow path guide 5a, the number of attachments, and the like are preferably set in consideration of the amount of the culture solution to be treated, the heating temperature, treatment time, ultraviolet irradiation time, and the like.

Further, the processing time of the processing apparatus 3 can be adjusted by controlling the pressure of a pump for feeding the culture solution in the purification apparatus 10 and the processing apparatus 3. The flow path guide 5a can be formed of a plate material or the like provided from the housing part 4a and / or the housing part 4b toward the flow path part 5. As a pump for feeding the culture solution, for example, a tube pump, a roller pump, a peristaltic pump or the like is used.

Next, with reference to FIG. 3A and FIG. 3B, a modified example of the processing apparatus 3 according to an embodiment of the present invention will be described. The processing apparatus 3 shown in FIGS. 3A and 3B includes the heating unit 6, the flow path unit 5, and the ultraviolet irradiation unit 8 (ultraviolet light source 8a) as one unit U, and two or more such units U are provided in parallel ( (Layered).
For example, as shown in FIG. 3A, a processing apparatus 3 in which only three units are provided in parallel can be used. According to this processing apparatus 3, since processing can be performed simultaneously with a plurality of units, it is possible to improve the processing capacity of virus inactivation processing (for example, if the number of units is two, the processing capacity is 2). Can be doubled, and if the number of units is three, the processing capacity can be tripled).

Further, in the processing apparatus 3 shown in FIG. 3B, an example in which four units U are provided is illustrated. Specifically describing the modification, the upper two units U in FIG. 3B are arranged with the

flow path portions

5 and 5 on the outer side of the ultraviolet irradiation unit 8 (ultraviolet light source 8a) and further on the outer side. The

heating units

6 and 6 are arranged. That is, the two units U are configured in such a manner that the ultraviolet irradiation unit 8 is shared between the units. And the lower two units U in FIG. 3B have the same configuration. In addition, in the center two unit U in FIG. 3B, it is comprised in the format which shares the heating part 6 arrange | positioned between the upper flow path part 5 and the lower flow path part 5. FIG. Thus, if it is the processing apparatus 3 shown to FIG. 3B, since it becomes the structure which shares the heating part 6 and the ultraviolet irradiation part 8 by several units U (two units U), size reduction of an apparatus is aimed at. be able to. Moreover, according to this processing apparatus 3, since the heating part 6 and the ultraviolet irradiation part 8 are shared by the some unit U, a heating and ultraviolet irradiation can be performed simultaneously. Therefore, according to this processing apparatus 3, not only can virus inactivation processing be performed efficiently, but also energy saving can be achieved.

Next, another modification of the processing apparatus 3 according to an embodiment of the present invention will be described with reference to FIG. The processing apparatus 3 shown in FIG. 4 is configured so that the flow path portion 5 includes a flexible bag 11 and

housing portions

4 a and 4 b that cover the bag 11. A flow path guide 5a for closing the part of the bag 11 and guiding the flow direction of the culture solution is provided inside the housing part 4a. In FIG. 4, as an example of the flow path guide 5a provided inside the housing portion 4a, the flow guide 5a is provided in the form shown in FIG. 2B, that is, the flow of the culture solution is meandering. Needless to say, it can be provided in different forms. Moreover, in this modification, in order to enable the bag 11 to be taken in and out of the

housing portions

4a and 4b, the bag 11 can be opened and closed by a hinge or the like. With such a configuration, when the bag 11 is put in the

housing parts

4a and 4b, the

housing parts

4a and 4b are closed and the bag 11 is sandwiched, the flow path guide 5a can form a flow path. .

As the flexible bag 11, for example, a single-use bag 11 for pharmaceutical packaging that is made of a multilayer film of ethylene / vinyl / acetate or ethyl / vinyl / alcohol can be used. Such a single-use bag 11 is sterilized in advance by gamma rays, ultraviolet rays, ethylene oxide gas or the like and maintained in a sterile state, and the contents such as gas and liquid are almost removed and provided in a clean state. It can use suitably for invention. In addition, when such a single-use bag 11 is used, a sterilization and washing process is not necessary as compared with the case where the stainless steel or glass flow path section 5 is adopted. Ancillary facilities such as water can be dispensed with. Therefore, the operability can be simplified and the apparatus can be simplified, and the operation cost can be reduced.

The processing apparatus 3 according to the present embodiment described above and the purification apparatus 10 including the processing apparatus 3 include the heating unit 6 that heats the culture solution and the ultraviolet irradiation unit 8 that irradiates the culture solution with ultraviolet rays. In addition, since the heat treatment and the ultraviolet irradiation treatment are simultaneously performed, the virus inactivation treatment can be efficiently performed instead of the conventional low pH treatment.

[Treatment method for cultured products]
Next, a method for treating a cultured product according to this embodiment (sometimes simply referred to as “treatment method” in the present specification) will be described. The processing method according to the present embodiment processes the culture product using the processing apparatus 3 described above. Since the flow path section 5, the heating section 6, the ultraviolet irradiation section 8, and the like, which are constituent elements of the processing apparatus 3, are as described above, description thereof is omitted. In the description of the processing method, the same components as those described in the processing device 3 are denoted by the same reference numerals, and the description thereof is omitted.

In the treatment method according to this embodiment, cells are first cultured in the culture tank 1 to obtain a culture solution containing a culture product. Next, at least cells are removed from the culture solution using a cell disruption unit, a filtration filter, a continuous centrifuge, or the like. And the culture solution from which the cell was removed and containing the culture product is stored in the 1st storage tank 2a. Next, the culture solution is transferred from the first storage tank 2a to the processing apparatus 3, and the virus inactivation process is performed by simultaneously performing the heating by the heating unit 6 and the ultraviolet irradiation by the ultraviolet irradiation unit 8. The culture solution on which the virus inactivation process has been performed is washed by the chromatograph 9 and stored in the second storage tank 2b.

Here, in the treatment method according to the present embodiment, the heating temperature of the culture solution by the heating unit 6 is preferably set to a temperature that does not cause thermal denaturation of the culture product. The temperature that does not cause the heat denaturation of the cultured product varies depending on the cultured product, and can be appropriately set according to the cultured product. For example, it can be set to 80 ° C. or the like, but may be set to 75 ° C. or 65 ° C. You can also. When the upper limit of the heating temperature of the culture solution by the heating unit 6 is set as described above, for example, even when the culture product is an antibody protein, heat denaturation is performed while inactivating the virus in the culture solution. It is possible to avoid such a situation that the activity is impaired by. In addition, it is preferable that the minimum of the heating temperature of the culture solution by the heating part 6 shall be 60 degreeC, for example.

Further, in the treatment method according to the present embodiment, it is preferable that the residence time of the culture solution from the inlet 5b to the outlet 5c of the flow path unit 5 is 30 seconds or longer and 30 minutes or shorter. In this way, reliable virus inactivation processing can be performed. In addition, from the viewpoint of more reliable virus inactivation treatment and efficient treatment, the residence time is preferably 1 minute or more and 20 minutes or less.

Furthermore, in the processing method according to the present embodiment, it is preferable that the wavelength of ultraviolet light is 254 nm. If it does in this way, inactivation of a virus can be performed effectively. Note that the wavelength of the ultraviolet light is not limited to this, and ultraviolet light having a shorter wavelength and ultraviolet light having a longer wavelength can be used as long as they fall within the so-called ultraviolet light.

As described above, the treatment method according to the present embodiment simultaneously performs the heat treatment and the ultraviolet irradiation treatment on the culture solution using the treatment apparatus 3 described above, and therefore performs virus inactivation treatment instead of the conventional low pH treatment. It can be done efficiently.

Although the culture product processing apparatus, the culture product processing method, and the culture product purification apparatus according to an embodiment of the present invention have been described in detail above, the gist of the present invention is not limited thereto. Various modifications are included. For example, the channel guide 5a may be formed in the bag 11 in advance by welding or the like.

The above-described embodiment has been described in detail for easy understanding of the present invention, and is not necessarily limited to one having all the configurations described. Further, a part of the configuration of an embodiment can be replaced with the configuration of another embodiment, and the configuration of another embodiment can be added to the configuration of an embodiment. In addition, it is possible to add, delete, and replace other configurations for a part of the configuration of each embodiment.

10 Purification equipment (culture product purification equipment)
DESCRIPTION OF SYMBOLS 1 Culture tank 2a 1st storage tank 2b 2nd storage tank 3 Processing apparatus (processing apparatus of culture product)

4a Housing part

4b Housing part 5 Flow path part 5a Flow path guide 5b Inlet 5c Outlet 6 Heating part 7 Cooling part 8 Ultraviolet irradiation part 8a Ultraviolet source 8b Ultraviolet reflection part 9 Chromatograph

Claims

A flat channel section having an inlet and an outlet through which a culture solution containing a culture product is removed and from which cells have been removed;
A heating section that is provided in contact with or close to one surface of the flow path section, and heats the culture solution;
A culture production comprising: an ultraviolet irradiation unit that irradiates the culture solution with ultraviolet rays, provided on the other surface of the flow path part in contact with or in proximity to a position facing the heating unit Material processing equipment.
The culture product treatment apparatus according to claim 1, wherein the heating temperature of the culture solution by the heating unit is equal to or lower than a temperature that does not cause thermal denaturation of the culture product.
The apparatus for treating a cultured product according to claim 2, wherein the temperature at which the cultured product does not undergo thermal denaturation is 80 ° C.
The culture product processing apparatus according to claim 1, wherein a residence time of the culture solution from the inflow port to the outflow port is 30 seconds to 30 minutes.
The apparatus for treating a cultured product according to claim 1, wherein the wavelength of the ultraviolet light is 254 nm.
The culture product processing apparatus according to claim 1, wherein a flow path guide for guiding a flow direction of the culture solution is provided in the flow path section.
The culture product processing apparatus according to claim 1, wherein the heating unit, the flow path unit, and the ultraviolet irradiation unit are provided as one unit, and two or more of the units are provided in parallel.
The flow path portion has a flexible bag, and a housing portion that covers the bag,
The culture product processing apparatus according to claim 1, wherein a flow path guide that closes a part of the bag and guides a flow direction of the culture solution is provided inside the housing portion. .
The culture product treatment apparatus according to claim 1, wherein the culture product is an antibody protein.
A flat channel section having an inlet and an outlet through which a culture solution containing a culture product is removed and from which cells have been removed;
A heating section that is provided in contact with or close to one surface of the flow path section, and heats the culture solution;
On the other surface of the flow path part, provided in contact with or close to the position facing the heating part, an ultraviolet irradiation part for irradiating the culture solution with ultraviolet light,
A method for treating the culture product using a culture product processing apparatus comprising:
A method for treating a cultured product, comprising: irradiating the culture solution in the flow channel from the ultraviolet irradiation unit while heating the culture solution in the flow channel by the heating unit.
The culture product treatment method according to claim 10, wherein the heating temperature of the culture solution by the heating unit is equal to or lower than a temperature that does not cause thermal denaturation of the culture product.
The method for treating a culture product according to claim 11, wherein the temperature at which the culture product does not cause thermal denaturation is 80 ° C.
The method for treating a cultured product according to claim 10, wherein a residence time of the culture solution from the inlet to the outlet is 30 seconds or longer and 30 minutes or shorter.
The method for treating a cultured product according to claim 10, wherein the wavelength of the ultraviolet ray is 254 nm.
An apparatus for purifying a culture product produced by culturing cells.
A flat channel portion having an inlet and an outlet through which the culture solution containing the culture product is passed and from which the cells have been removed, and one surface of the channel portion are provided in contact with or close to each other A heating unit that heats the culture solution, and is provided in contact with or in proximity to a position facing the heating unit on the other surface of the flow path unit. An apparatus for treating a cultured product, comprising: an ultraviolet irradiation unit that irradiates ultraviolet rays together with heating;
A culture product purification apparatus comprising: a chromatograph provided at least before and after the culture product processing apparatus with reference to the flow direction of the culture solution.