JP6817677B1 - Bioreactors, bioreactor manufacturing methods, and disposable bags used in bioreactors - Google Patents

Abstract

PROBLEM TO BE SOLVED: To have a problem that a conventional SUS bioreactor is expensive for sterilization equipment and a single-use bioreactor is also expensive. A bioreactor of the present invention sterilizes a disposable bag having an opening and containing contents, a reusable lid device that seals and closes the opening of the bag, and the inside of the bag. It comprises a sterilization means, and the lid equipment comprises a lid body and a bioreactor equipment provided on the lid body and inserted into the bag. Further, the sterilization means is sterilization by irradiation with ultraviolet rays. It also has a storage container for fixing the bag. The lid equipment is made of stainless steel. [Selection diagram] Fig. 1

Description

The present invention relates to a bioreactor, particularly a bioreactor for culturing microorganisms and molds.

For example, regarding a bioreactor (biological reaction device) that cultivates microorganisms and molds while stirring in a culture vessel, the bioreactor equipment that constitutes this bioreactor and the stirring blades and pipes that accompany it are , Mainly made of stainless steel (SUS) is used.

The sterilization method of this stainless steel bioreactor is generally high temperature steam sterilization (SIP) and chemical washing sterilization (CIP), and a pressure vessel is required for steam operation.

Furthermore, in order to prevent contamination, it is necessary to sterilize all SUS pipes, various utilities such as steam are required, and various equipment other than the bioreactor is required.

For this reason, the cost of the incubator is high, the operation is troublesome, and the risk of operation error is high.

Therefore, in animal cell culture and the like, a single-use bioreactor that solves these problems and can be operated even in a simple room without a utility as described above has been commercialized and has become common.

This single-use bioreactor does not have a SUS tank, but a flexible bag such as a disposable (single-use) bag made of flexible synthetic resin (plastic such as polyethylene, elastomer) made of flexible material. It is used, and a bioreactor facility is integrally provided in the bag, and the whole is disposable.

By sterilizing this bag in advance (γ sterilization) and making it disposable with a structure that keeps the inside of the bag sealed, it is not necessary to construct a complicated structure or sterilization system such as a SUS reactor.

In addition, the piping provided in the bag can be aseptically connected with a tube made of a flexible material, which reduces the risk of contamination and makes operation extremely easy.

For example, as a single-use bioreactor, there is, for example, Patent Document 1.

Japanese Patent Application Laid-Open No. 2016-212017 No. 2

However, such disposable sealed bioreactors are very expensive, and if the added value of the product is high, it is profitable, but the added value of the product is low, and microorganisms that require low cost and mass production It was difficult to operate in mold culture.

In addition, when a manufacturer newly enters the biotechnology business, a large amount of development cost is required to consider PILOT with a final production capacity of several tens of tons to 200 tons, and investment in SUS equipment has become a bottleneck.

The present invention provides a PILOT single-use bioreactor that enables microbial and fungal culture at low cost.

In order to achieve the above object, the bioreactor of the present invention includes a bag-shaped disposable bag having an opening for containing the contents, and a reusable lid facility for sealing and closing the opening of the bag. It consists of a sterilizing means for sterilizing the inside of the bag.
The disposable bag is not provided with bioreactor equipment required for processing the contents , and the lid equipment is provided on the lid body and the lid body and inserted into the bag to provide the contents. characterized by comprising further a necessary by Ru bioreactor equipment object of processing.

Further, the sterilizing unit state, and are sterilized by ultraviolet irradiation, characterized that you have provided to the lid body.

Further, a storage container having an open upper end for storing and fixing the bag is further provided , a collar portion is formed on the entire circumference of the upper end opening edge of the storage container, and the entire opening edge of the bag is formed. circumferentially, the flange portion of the container, characterized that you have flange portion is formed to be sandwiched between the lower surface of the lid body.

Further, the lid equipment is made of stainless steel.

Further, the disposable bag used in the bioreactor of the present invention comprises a bag-shaped disposable bag in which the opening is hermetically closed by a sealing member to accommodate the contents, and the disposable bag is the content. The bioreactor equipment required for the treatment of the above is not provided, and after the sealing member is removed, it is inserted into the lid body and the bag provided on the lid body, and is required for the treatment of the contents. a more becomes a lid and facilities bioreactor equipment are, opening of the bag, characterized in Rukoto closed hermetically.

Further, in the method for manufacturing a bioreactor of the present invention, the sealing member is removed from a bag-shaped disposable bag in which the contents are contained, and the sealing member is sealed and closed, and the bag is opened. A first step, a second step of sealing and fixing the reusable lid equipment to the opening of the opened bag, and a third step of sterilizing the inside of the sealed bag by sterilization means. Therefore, the disposable bag is not provided with the bioreactor equipment required for processing the contents, and the lid equipment is provided on the lid body and the lid body and inserted into the bag . characterized by comprising further a necessary by Ru bioreactor equipment processing the contents.

Further, the sterilizing unit state, and are sterilized by ultraviolet irradiation, characterized that you have provided to the lid body.

Further, in the second step of sealing and fixing the lid equipment to be reused in the opening of the opened bag, the bag is inserted into a storage container having an open upper end and stored , and the bag is opened. a flange portion formed on the entire circumference of the edge, a flange portion formed on the entire circumference of the upper end opening edge of the container, and wherein Rukoto such than to fix by sandwiching by the lower surface of the lid body To do.

Further, the lid equipment is made of stainless steel.

According to the present invention, it becomes possible to provide a bioreactor having a simple structure and a low cost.

It is explanatory drawing which shows the whole view of the bioreactor of this invention. It is explanatory vertical sectional view of the disposable bag used for the bioreactor of this invention. It is explanatory drawing of the manufacturing method of the bioreactor of this invention. It is explanatory drawing of the manufacturing method of the bioreactor of this invention. It is explanatory drawing of the manufacturing method of the bioreactor of this invention.

Examples of embodiments for carrying out the present invention are shown below.

Example 1 of the present invention will be described with reference to FIGS. 1 to 5.

As shown in FIG. 1, the bioreactor of the present invention includes a bag-shaped disposable bag (single-use bag) 1 having an open upper end, which contains contents composed of a culture such as microorganisms and molds and a culture solution. , The bag 1 is erected and stored and fixed, and the storage container 2 and the opening at the upper end of the bag 1 are sealed (airtight, liquidtight) and fixed, and reused (reused) by sterilization. It is composed of, for example, a stainless steel (SUS) lid equipment 3 which is used), and the lid equipment 3 is provided on the lid main body 4 which is hermetically fixed to the upper end of the bag 1 and the lid main body 4. It comprises a bioreactor facility 5 required for processing the contents in the bag 1 which is inserted into the bag 1.

The disposable bag 1 is sterilized before use (for example, EOG sterilized, γ sterilized), and after use, it is discarded without being washed and reused, and another disposable bag 1 is used for the next use. Is newly provided and used.

The bioreactor facility 5 includes, for example, a stainless steel stirring means 6 including a stirring shaft 6a and a stirring blade 6b that stirs the contents in the bag 1, and supplies / discharges liquid / gas inside / outside the bag 1. There are stainless steel pipes 7 for heating and cooling the contents, spudgers, baffles (not shown), and the like.

Further, the storage container 2 is made of, for example, a bottomed cylinder having an open upper end, and is placed or fixed on a floor 13 or a mounting table, etc., and a stored flexible bag 1 is fixed to the bioreactor. To be able to process.

The bag 1 is, for example, a flexible synthetic resin bottomed tubular container having an open upper end, and the upper end opening of the bag 1 is in a state before the lid equipment 3 is fixed. In, as shown in FIG. 2, a film-like sealing member such as a sterilized film 8 is formed in the bag 1 in a state in which a gas is sealed and inflated so that the inside of the bag 1 is not contaminated. It is closed, fixed, and sealed.

Then, when the bag 1 is used, the sterilizing film 8 is removed from the bag 1 so that the lid equipment 3 can be attached.

A collar portion 9 extending outward in the radial direction is formed on the entire circumference of the upper end opening edge 1a of the bag 1, and a flange portion 9 extending outward in the radial direction is formed on the entire circumference of the upper end opening edge 2a of the storage container 2. An extending collar portion 10 is formed, and the collar portion 9 of the bag 1 is sandwiched between the collar portion 10 of the storage container 2 and the lower surface of the lid body 4, so that the inside of the bag 1 is sealed. ..

Further, the capacity of the bag 1 is, for example, about 10L to 2000L, but the capacity can be as large as possible.

The lid equipment 3 is preferably made of stainless steel (SUS) so that it can be sterilized by sterilization means and reused, but there is no particular limitation as long as the lid equipment 3 is a reusable metal.

Further, the lid body 4 is provided with a sterilization means 11 for sterilizing the inside of the bag 1. As the sterilization means 11, for example, there is an ultraviolet irradiation device such as an ultraviolet lamp, and the ultraviolet irradiation device is provided so as to penetrate the lid body 4 and is formed so that the bag 1 can be irradiated with ultraviolet rays. .. The irradiated ultraviolet rays are preferably deep ultraviolet rays (wavelength: 200 nm to 300 nm), and in particular, deep infrared rays having a wavelength of around 265 nm are easily absorbed by DNA, and the effect of losing genetic information has been confirmed. preferable.

Further, the storage container 2 is provided with a sterilization means 12 for sterilizing the inside of the bag 1. As the sterilization means 12, for example, there is an ultraviolet irradiation device such as an ultraviolet lamp, and the ultraviolet irradiation device is provided on, for example, the peripheral surface or the bottom surface of the storage container 2 so that the bag 1 can be irradiated with ultraviolet rays. Form in. The irradiated ultraviolet rays are preferably deep ultraviolet rays (wavelength: 200 nm to 300 nm), and in particular, deep infrared rays having a wavelength of around 265 nm are easily absorbed by DNA, and the effect of losing genetic information has been confirmed. preferable.

In such a case, the bag 1 is made of a material through which ultraviolet rays pass, or is transparent or translucent.

Reference numeral 14 denotes a culture control device, which controls the flow of liquid or gas in the stirring means 6 or the pipe 7 to culture the contents in the bag 1. Control.

The storage container 2 may be omitted.

Further, the sterilization means 11 and the sterilization means 12 may be only one of them. Further, it may be separated from the lid main body 12 or the storage container 2.

Next, the method and effect of producing the bioreactor of the present invention will be described.

In the bioreactor of the present invention, the single-use bag 1, the storage container 2, and the lid equipment 3 are separately provided before use.

The single-use bag 1 is sterilized (for example, EOG sterilized, γ sterilized) and is inexpensive.

Further, before use, the sterilization film 8 is attached to the bag 1.

Then, in order to assemble the bioreactor, first, the sterilizing film 8 is removed from the single-use bag 1, the upper end is opened, and the bag 1 is inserted into the storage container 2, and the collar portion 9 of the bag 1 is inserted into the storage container 2. The collar part 10 of the

Then, the lid equipment 3 is inserted so that the bioreactor equipment 5 enters the bag 1, and the outer peripheral portion of the lower surface of the lid main body 4 and the flange portion 10 of the storage container 2 are used to form the bag 1. By sandwiching the collar portion 9 and fixing the collar portion 10 of the storage container 2 and the lid body 4, the bag 1 and the lid equipment 3 are hermetically fixed.

When the storage container 2 is not used, the collar portion 9 of the bag 1 and the lid body 4 are fixed.

At the time of this assembly, that is, when the sterilization film was peeled off, the bag 1 was once non-sterilized and the inside of the bag 1 which was a sterilized environment may be contaminated, but the bag 1 and the lid equipment 3 were fixed. Later, the inside of the bag 1 is irradiated with ultraviolet rays for a certain period of time by the sterilization means 11 and 12, and sterilization is performed.

Such ultraviolet irradiation is sterilized for, for example, half a day to one day.

Then, the contents are used for culturing or the like in the sterilized bag 1.

After use, the lid equipment 3 is removed from the bag 1, only the bag 1 is discarded, and the lid equipment 3 with the incidental equipment of the bioreactor device 5 is sterilized for reuse. Attach it to a new bag 1 and move on to the next process.

For example, after use, the lid equipment 3 is lifted by an elevating device or the like to remove the bag 1, the elevating device is washed, and then lowered to seal a new bag 1 with a seal or the like. ..

The bag 1 may be stored in the storage container 2 before the sterilization film is removed, or after the lid equipment 3 is attached to the bag 1.

According to the present invention, only the bag 1 in the bag-shaped portion is disposable, the lid equipment with ancillary equipment is reused, and the contamination at the time of installation of the bag 1 and the lid equipment 3 is caused by the ultraviolet irradiation. By performing in-tank sterilization, it becomes possible to provide a bioreactor having a simple structure and low cost.

In addition, the disposable bag 1 is simply a bag shape, has no connection such as a tube, is already sterilized (for example, EOG sterilization, γ sterilization), is inexpensive, and does not require sterilization, so the structure is simple and low cost bioreactor. It will be possible to provide a reactor.

Furthermore, since the culture preparation is simplified, the load on the operator is reduced.

In addition, the bioreactor can be installed anywhere and can be operated in a normal office, eliminating the need for equipment costs other than the bioreactor.

The bioreactor of the present invention is used for stirring in medical products, foods, and the like.

1 Single-use bag 1a Top opening edge 2 Storage container 2a Top opening edge 3 Lid equipment 4 Lid body 5 Bioreactor equipment 6 Stirring means 6a Stirring shaft 6b Stirring blade 7 Piping 8 Sterilization film 9 Flange 10 Flange 11 Sterilization means 12 Sterilization Means 13 Floor 14 Culture control device

Claims (9)

A bag-shaped disposable bag with an opening that can hold the contents,
Reused lid equipment that seals and closes the opening of the bag,
It consists of a sterilization means for sterilizing the inside of the bag.
The disposable bag is not equipped with the bioreactor equipment required to process the contents.
Lid facility, a lid body, provided in the lid body, said inserted into the bag, a bioreactor characterized by comprising more required and is Ru bioreactor equipment processing the contents. The sterilizing means may state, and are sterilized by UV irradiation, bioreactor according to claim 1, characterized that you have provided to the lid body. It also has a storage container with an open top for storing and fixing the bag .
A collar portion is formed on the entire circumference of the upper end opening edge of the storage container, and a collar portion sandwiched between the collar portion of the storage container and the lower surface of the lid body is formed on the entire circumference of the opening edge of the bag. the bioreactor of claim 1 or 2 is formed, characterized in Rukoto. The bioreactor according to claim 1, claim 2 or claim 3, wherein the lid equipment is made of stainless steel. It consists of a bag-shaped disposable bag that can hold the contents, with the opening sealed and closed by a sealing member.
The disposable bag is not equipped with the bioreactor equipment required to process the contents.
After the sealing member is removed, the bag is opened by a lid facility that is inserted into the lid body and the bag provided on the lid body and comprises a bioreactor facility required for processing the contents. disposable bag for use in bioreactor section is characterized Rukoto closed hermetically. The first step of removing the sealing member from the bag-shaped disposable bag containing the contents and opening the bag, in which the sealing member is sealed and closed in the opening.
A second step of sealing and fixing the reusable lid equipment to the opening of the opened bag,
It consists of a third step of sterilizing the inside of the sealed bag by sterilization means.
The disposable bag is not equipped with the bioreactor equipment required to process the contents.
The lid facility, a lid body, provided in the lid body, is inserted into the bag, a manufacturing method of the bioreactor characterized by comprising more necessary and are Ru bioreactor equipment processing the contents. The sterilizing means may state, and are sterilized by ultraviolet irradiation, the production method of the bioreactor according to claim 6, characterized that you have provided to the lid body. The second step of sealing and fixing the reusable lid equipment to the opening of the opened bag is
The bag is inserted into a storage container with an open upper end and stored .
A flange portion formed on the entire circumference of the opening edge of the bag, and a flange portion formed on the entire circumference of the upper end opening edge of the container, that Do than be fixed by sandwiching by the lower surface of the lid body The method for producing a bioreactor according to claim 6 or 7, wherein the bioreactor is manufactured. The method for manufacturing a bioreactor according to claim 6, claim 7 or claim 8, wherein the lid equipment is made of stainless steel.

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