JP2003047457A - Installation for producing vector and method for producing vector - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an installation for producing a vector, satisfying the physical containment level P2 or P3 specified in the recombinant DNA experimental standard, and the safety standard specified by GMP, and having excellent productivity, and further to provide a method for producing the vector. SOLUTION: This installation for producing the vector has an anteroom 3, a first pre-airlock room 5, a vector-producing room 2, a post-airlock room 7 and a sterilization treatment room 8. Doors are installed between respective room so as to form the line of flow restricted so that an operator may enter the anteroom 3, may move the first pre-airlock room 5, the vector-producing room 2 and the post-airlock room 7 in order and may go out through the sterilization treatment room 8 to the exterior. The installation has a room pressure- regulating means for regulating the pressure in each of the rooms.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a vector production facility and a vector production method used for gene therapy. More specifically, the physical containment levels P3 and GMP (Good Ma
nufacturing Practice) The present invention relates to a vector production facility and a vector production method that satisfy the containment level defined in the standard.

[0002]

2. Description of the Related Art In recent years, research and clinical development of disease treatment using therapeutic genes have been advanced, and great expectations are placed on the results. For this gene therapy, a gene recombinant called a vector that plays a role of carrying a therapeutic gene is used.

Vectors are often produced by introducing a therapeutic gene into a virus. Originally, a virus has the property of releasing a gene inside the virus when it infects a cell. Therefore, when a vector in which a therapeutic gene is introduced into a virus is used, the therapeutic gene introduced into the virus is released when the vector virus infects a cell to be treated.
Therefore, the effect of gene therapy by the vector can be exerted.

However, since the virus originally has pathogenicity, a treatment for removing pathogenicity is necessary in order to use the virus as a vector. Therefore, when a virus is used as a vector, processing for removing the gene body in the virus or processing for removing the gene that activates the gene body in the virus is performed.

[0005] Such a virus-based vector can be prepared by removing the pathogenicity of the virus, preparing the therapeutic gene, introducing the therapeutic gene into the virus from which the pathogenicity has been removed, and treating the virus. It is necessary to culture the vector into which the gene for use has been introduced. In addition to this, quality control such as storage of materials used for vector production, storage of produced vector and confirmation of performance is also required.

[0006] A particularly important control item in preparing a vector is ensuring safety. For the production of vectors, it is necessary to confirm that dangerous viruses are handled, that a new type of product called a genetically modified product is generated, and that these materials and products, the vectors, are not visible to the naked eye. There is a danger that it will not be easy. Therefore, unless measures are taken against these risks, vector preparation and experiments cannot be performed. In other words, it is important to prevent the transmission of dangerous substances to the creators and experimental workers and the diffusion to the outside world, and the transmission and diffusion of the genetically modified substance (vector) to the environment.

Regarding the assurance of safety in recombinant DNA experiments, there is "Recombinant DNA Experiment Guideline in Universities" announced by the Ministry of Education. In addition, GMP (Good Manufact
uring Practice). In creating vectors, at least the physical containment criteria specified in these guidelines and criteria must be met.

Regarding the above physical containment criteria,
In the "Recombinant DNA Experiment Guideline for Universities", four levels from P1 to P4 are shown for facilities and work. The main criteria are, for example, at the P2 level, when using a device that tends to generate a large amount of aerosol,
Make sure that contaminated aerosols do not leak outside, keep the laboratory in a building equipped with a high-pressure steam sterilizer, sterilize all waste before disposal, and wear lab coat in the laboratory. , Take off when you leave. At the P3 level, in addition to the P2 level, a safety cabinet should be installed, an experimental area separated by an antechamber to control entrance and exit should be provided, and the front and rear doors should not be opened at the same time. , It is regulated that the air flow is from the antechamber to the experimental area.

The safety standards in GMP are that the building must meet at least the P2 level mentioned above, that the workers before the vector preparation and the raw materials / equipment necessary for the vector preparation and the workers after the vector preparation are contaminated. It is stipulated that the waste does not come into contact with the product vector.

As described above, facilities and operations in experiments for gene recombination and vector preparation require sufficient safety measures and measures to prevent contamination of foreign substances, and some proposals have been made as such measures. Has been done. For example, Japanese Unexamined Patent Publication No. 6-82024 discloses a method for enhancing the safety of exhaust treatment. In this method, measures are taken to ventilate the exhaust gas from the facility through a filter and then heat (burn) the exhaust gas. In addition, JP-A-6-241
Japanese Patent No. 519 discloses an alarm device for a safety cabinet that can prevent occurrence of biohazard by notifying an experimenter of abnormality of the safety cabinet by a pressure switch and an alarm device.

The measures disclosed in these publications are safety measures for some devices such as laboratories, and these techniques are described in the above-mentioned "Recombinant D in Universities".
It only contributes to a small part for constructing a vector production facility that satisfies the safety standards stipulated in “NA experimental guidelines” and GMP.

[0012]

DISCLOSURE OF THE INVENTION The present invention provides a recombinant DN
Physical containment level P2 specified in A Experiment Guideline
Or P3 and GMP (Good Manufacturing Practic
It is an object of the present invention to provide a vector production facility and a vector production method which are advantageous in satisfying the safety standard for containment defined in e) and have excellent productivity.

[0013]

The gist of the present invention lies in the following vector production facility and vector production method.

The vector production facility according to the present invention comprises a preparation room, a first pre-airlock room, a vector preparation room, a post-airlock room, and a sterilization room. 1 air lock pre-chamber, the vector preparation chamber and the air lock post-chamber are moved in order to form a restricted flow line of going out through the sterilization chamber. A door is provided, and an indoor pressure adjusting unit for adjusting the indoor pressure of each room is further provided.

In the above-mentioned vector preparation facility, a preparation room is provided with a first preparation room, a second preparation room, and a second pre-airlock provided between the first preparation room and the second preparation room. The first preparation room, the second pre-airlock room,
It is desirable that a door be provided between the rooms so as to form a restricted flow line of moving in the order of the second preparation room.

Further, these vector production facilities preferably have a changing room outside the entrance door of the preparation room or the first preparation room, and a dressing room outside the exit door of the sterilization room. Is more desirable.

The room pressure of each room in the above-mentioned vector production facility is lower or higher than that in the preparation room and the vector production room in the first airlock chamber, and is lower than that in the vector production room and the sterilization chamber after the airlock. It is desirable that the temperature is higher than or equal to or higher than the atmospheric pressure and that the vector production chamber and the sterilization chamber are maintained lower than the atmospheric pressure.

Further, the room pressure in each room is equal to the second room pressure.
The airlock front chamber is lower or higher than the first preparation chamber and the second preparation chamber, the first pre-airlock chamber is lower or higher than the second preparation chamber and the vector preparation chamber, and the airlock rear chamber is Is lower or higher than the vector preparation room and the sterilization room, and it is desirable that the vector preparation room and the sterilization room are maintained below atmospheric pressure.

Further, it is desirable to provide a vector storage room in which the installation position and the position of the door are determined so that the worker can enter and exit without going through the preparation room or the first preparation room. It is desirable to provide a quality control room in which the installation position and the position of the door are determined so that the person can enter and exit without going through the first preparation room.

The vector production method of the present invention comprises a preparation room, a first pre-airlock room, a vector preparation room, a post-airlock room, and a sterilization room, where an operator prepares the preparatory room, the first room.
To move each chamber in the order of the airlock front chamber, the vector preparation chamber and the airlock rear chamber so as to form a restricted flow line of going out of the facility through the sterilization chamber,
A door is provided between the rooms, and the indoor pressure is controlled by the vector preparation facility equipped with the indoor pressure adjusting means for adjusting the indoor pressure of each room. Lower or higher, the post-airlock chamber is lower or higher than the vector preparation room and the sterilization room, and the vector preparation room and the sterilization room are maintained at a pressure lower than atmospheric pressure. Characterize.

Another method for producing a vector of the present invention is
A first preparation room, a second pre-airlock room, a second preparation room, a first pre-airlock room, a vector preparation room, a post-airlock room and a sterilization processing room It is said that the preparation room, the second pre-airlock room, the second preparatory room, the first pre-airlock room, the vector preparation room, and the post-airlock room are moved in that order, and the room goes out through the sterilization room. A door is provided between the rooms so as to form a restricted flow line, and the indoor pressure is controlled by the vector production facility equipped with an indoor pressure adjusting means for adjusting the indoor pressure of each room. The airlock front chamber is lower or higher than the first preparation chamber and the second preparation chamber, the first airlock front chamber is lower or higher than the second preparation chamber and the vector preparation chamber, and the airlock rear chamber is Lower or higher than vector production room and sterilization room, and Over produced chamber and sterilization chamber while maintaining lower than the atmospheric pressure, and performing vector production work.

The "door" provided between the rooms in the present invention means a door such as a sliding door or a hinged door that can be partitioned. Further, the indoor pressure adjusting means is mainly a device such as a blower capable of supplying or exhausting air, a device for adjusting the amount of supplied air or the amount of exhausted air, an indoor pressure configured by an air supply or exhaust duct and a pressure detector. Means a means capable of adjusting.

According to the vector production facility or vector production method of the present invention described above, since the flow lines of the operator, the raw materials and the tools necessary for vector production are limited in one direction, it is possible to prepare before and after vector production. There is no contact with subsequent workers. In addition, there is no risk that the air in the vector production room leaks to the outside, and all wastes and exports can be sterilized by a high-pressure steam sterilizer before being exported to the outside. As described above, since contaminated air and contaminants do not leak outside from the vector production facility, the physical containment level P2 or P3 and GMP specified in the recombinant DNA experimental guideline are defined. It is advantageous in fully satisfying the safety standards regarding containment. Furthermore, it has the advantage of being highly productive because the flow line in the facility is limited to one direction and each room can be rationally arranged.

[0024]

BEST MODE FOR CARRYING OUT THE INVENTION FIG. 1 shows an arrangement example of each room in a vector production facility according to an embodiment of the present invention. Figure 1
As shown in FIG. 2, the vector production facility according to the embodiment of the present invention includes a preparation room 3, a first pre-airlock room 5,
A vector preparation room 2, an airlock post-chamber 7, and a sterilization processing room 8 are provided. Each of these chambers is a room that is indispensable for the production of a vector, and in addition, a changing room 10 is provided on the inlet side of the preparation room 3, a dressing room 11 is provided on the outlet side of the sterilization chamber 8, a sterilization chamber 8 and a partition wall. It is desirable to provide a vector storage room 12 with a space between them, a quality control room 13 with a partition wall from the vector storage room, and a control room 14 for monitoring and controlling the entire facility. It is desirable that each of these rooms be housed in one building.

When a vector is produced in this facility, the operator, the raw materials, and the equipment necessary for producing the vector enter the preparation room 3, and the first pre-airlock chamber 5, the vector production room 2 and The airlock post-chamber 7 is moved in that order, and a restricted flow line is formed so as to flow in one direction through the sterilization chamber 8 to the dressing room. Also,
It is desirable for each room to be able to adjust the room pressure individually. Therefore, doors for partitioning each room are required on the entrance side and the exit side of each room, and as shown in FIG. 1, doors D1, D2, D3, D6, D7, D8, etc. Is provided. In addition, it is preferable that the entrance-side door and the exit-side door in each room are provided in the partition walls facing each other in each room or in the vicinity thereof.

Further, this facility is provided with the indoor pressure adjusting means for adjusting the indoor pressure of each room as described above. The indoor pressure adjusting means is composed of a blower, a duct, an air supply port, an exhaust port, an air pressure detector, and the like. It is desirable that the air supply port and the air exhaust port are provided in each chamber in order to accurately control the indoor pressure. However, as described below,
It does not necessarily have to be provided in all rooms.

The vector production facility and the vector production method according to the present invention having the above-mentioned configuration will be described in detail below.

The vector production facility shown in FIG. 1 is a facility according to a preferred embodiment of the present invention. The vector production facility shown in FIG. 1 is designed so that preparation for vector production, vector production, vector storage, quality control, etc. can all be performed within one building 1.
Furthermore, the workers and raw materials / products are designed to flow in one direction.

There is a changing room 10 inside the entrance 16. To the changing room 10, there is an entrance door D11 and an entrance door D9 of the changing room 10.
It is supposed to enter through. A preparation room 3 is provided next to the changing room 10 via a partition wall, and a door D1 is provided on the partition wall so as to be movable from the changing room 10 to the preparation room 3. Furthermore, the vector preparation chamber 2 is arranged apart from the preparation chamber 3 by a first airlock front chamber. Figure 1
2 shows the case where two vector production chambers 2 (2-1, 2-2) are provided, the number of vector production chambers may be one, or three or more.

Further, in FIG. 1, a first airlock front chamber 5-1 and a vector preparation chamber 2 for the vector preparation chamber 2-1 are shown.
2 shows an example in which the first airlock prechamber 5 is provided independently of the vector preparation chamber 2 such as the first airlock prechamber 5-2 for -2.
However, the first airlock front chamber is defined as one chamber and one first
From the airlock front room to the vector production room 2-1 and 2-2
It may be configured so that it can be moved to both chambers. In addition,
From the preparation room 3 to the vector production room 2, the door D2 (D2-
1, D2-2), first airlock anteroom and door D3
It is possible to move via (D3-1, D3-2).

Here, the preparation room is a room for storing raw materials and instruments necessary for vector production, preparation for vector production, pretreatment, pre-process and the like. Also,
The first airlock anteroom is a room for preventing the air in the vector preparation room where dangerous substances and contaminants may be suspended from leaking to the preparation room.

A post-airlock chamber 7 and a sterilization chamber 8 sandwiching the post-airlock chamber 7 are disposed downstream of the vector preparation chamber 2 in the process. The partition wall between the airlock rear chamber 7 and the vector production chamber 2 is provided with a door (D6) so as to be movable from the vector production chamber to the airlock rear chamber.
-1, D6-2) are provided. Further, a door D7 is provided between the airlock post-chamber 7 and the sterilization chamber 8 so that the door D7 can be moved between the chambers.

The sterilization chamber 8 is equipped with a high-pressure steam sterilizer 9 for performing a sterilization process before carrying it out so that dangerous substances and contaminants do not come out. The high-pressure steam sterilizer 9 has a structure in which a sterilizer can be loaded from the sterilization chamber 8 side and a sterilized product can be taken out of the sterilization chamber 8 such as the undressing chamber 11. .

A vector storage chamber 12 is preferably arranged outside the sterilization chamber 8 via a partition wall. Further, the partition wall is preferably provided with a product delivery port 15 for transferring the vector from the sterilization chamber 8 directly to the vector storage chamber 12. Further, it is desirable that a dressing room 11 is arranged outside the door D8 on the exit side of the sterilization processing room 8.

It is desirable that a quality control room 13 is disposed adjacent to the vector storage room 12, and a product delivery port is provided in a part of the partition wall between the two rooms. It is desirable that the vector storage room 12 and the quality control room 13 have their installation positions and door positions determined so that an operator can enter and exit without going through the preparation room.

FIG. 2 shows an arrangement example of each room in a vector production facility according to another embodiment of the present invention. The difference from the vector production facility shown in FIG. 1 is that the preparation room 3 is divided into a first preparation room 3-1 and a second preparation room 3-1. Between the second preparation room 3-2 and the second
The airlock front chamber 5 is arranged. First
From the preparation room 3-1 to the second preparation room 3-2, the door D2,
Moving through the second airlock anteroom and door D3,
From the preparation room 3-2 to the vector production room 2
1, through the second airlock front chamber 6-1 and the door D5-1 to the vector production chamber 2-1, or through the door D4-2, the second airlock front chamber 6-2 and the door D5-2 It is configured so that it can be moved to the vector production room 2-2.

In this way, the airlock front chamber is divided into the first and second chambers.
By adopting the two stages, it is possible to greatly suppress the possibility that a dangerous substance or the like leaks from the vector preparation room 2 to the changing room 10 side and further improve the safety. When two preparation chambers, a first preparation chamber and a second preparation chamber, are provided, the first preparation chamber is for storing raw materials and instruments, and the second preparation chamber is for vector production. Good to use as a preparation room.

The functions of the above-mentioned rooms are as follows. The changing room 10 is a room for changing into clothes for performing vector production work. When the first preparation room 3-1 and the second preparation room 3-2 are provided as the preparation room, the first preparation room 3-1 includes a virus and a gene for recombination necessary for producing a vector. The second preparation room 3-2 is a room where simple pretreatment is carried out. In the vector preparation room, which is a sterile room, it is necessary to minimize complicated processing.
The preparation room 3-2 is suitable as a processing room therefor. When the preparation room is not divided into the first preparation room and the second preparation room, one preparation room has both the functions of the first preparation room and the second preparation room. .

The vector preparation room 2 is a room for literally preparing a vector, in which processing such as elimination of virus pathogenicity, extraction of therapeutic gene, recombination with viral gene and the like are carried out. Since these processes usually need to be performed in a safety cabinet, the safety cabinet 15 is installed in the vector production chamber 2 as shown in FIG.
Is equipped with. This safety cabinet 15
The internal pressure can be controlled so as to be a negative pressure compared to the inside of the room, and it is considered that the air inside the safety cabinet 15 does not leak to the vector production room 2. Further, a filter having an action of capturing microorganisms is attached to an air exhaust system inside the cabinet, and a HEPA filter is suitable as a type of filter.

The production work of the vector can be carried out by putting the raw materials and instruments in the safety cabinet 15 and through the rubber gloves which can be reached from the vector production room side. Therefore, the risk of virus or vector leaking into the vector production room during the work is low. However, the vector production room is the most dangerous place because virus handling and vector culture are performed in the vector production room before and after the work. Measures for avoiding this risk, that is, measures for ensuring safety will be collectively described later.

Sterilization chamber The chamber 8 is a room for sterilizing the instruments used for vector production work and waste that may be contaminated. A virus or vector can be killed by exposing it to a temperature of usually about 0 ° C, and thus can be sterilized even by heat treatment at such a temperature. The autoclave treatment by the high-pressure steam sterilizer 9 is a treatment for further increasing the certainty of the sterilization treatment.
It is preferable to perform the treatment under the conditions of 2 atm and 121 ° C.

FIG. 1 shows a first pre-airlock chamber 5 and a vector production chamber 2 between the preparation chamber 3 and the vector production chamber 2.
After the airlock between the sterilization processing chamber 8 and the sterilization processing chamber 8, in FIG. 2, the preparation chamber 3 is replaced with the first preparation chamber 3-1 and the second preparation chamber 3-2, as compared with the case of FIG. An example is shown in which the second airlock front chamber 5 is arranged between the first preparation chamber 3-1 and the second preparation chamber 3-2 (hereinafter, referred to as an airlock front chamber and an airlock). The rear room is collectively referred to as the airlock room).

Each of these airlock chambers has a role as a barrier against the chambers before and after it. That is, the vector produced in the vector production room 2 and contaminants that may float in the air are transferred from the vector production room 2 to the preparation room 3 or the second preparation room 3-2, and the first preparation room 3-2. It plays a role of preventing leakage to the preparation room 3-1 or from the vector preparation room 2 toward the sterilization processing room 9. If there is a possibility that dangerous substances or contaminants may move from the vector preparation room 2 to the preparation room 3 or the first preparation room 3-1, the second preparation room 3-2, or the sterilization processing room 8. Even if a situation occurs, the air lock front room or the air lock back room
It is a room that works to block the flow.

The undressing room 11 is a room in which work clothes worn during work are changed. In addition, the vector storage room 12
The quality control room 13 is a room for temporarily storing a vector, which is a product, and is a room for performing a quality inspection of the vector and quality control in the manufacturing process. The control room 14 monitors the equipment of the entire vector production facility and the air pressure in each room,
It is a room for controlling so as to maintain a normal state, and serves as a central control room.

The vector production facility according to the present invention thus constructed has physical containment levels P2 and P3 and GMP (Good) defined in the recombinant DNA experimental guidelines.
Since it satisfies the safety standards for containment defined in (Manufacturing Practice), according to this facility,
The vector can be produced safely and with high productivity.
The security will be specifically described below.

The vector production facility according to the present invention employs three characteristic measures in order to ensure safety. As already mentioned, the first measure is that the flow lines of workers, raw materials, instruments and vectors such as products flow in one direction from the changing room on the entrance side to the changing room on the exit side. It is configured to. The second measure is that a barrier can be formed by providing a single or double airlock front chamber between the changing room and the vector preparation room and an airlock rear chamber between the vector preparation room and the undressing room. It is designed to. The third measure is configured so that the room pressure in each room from the changing room to the undressing room can be appropriately controlled so that dangerous substances do not leak out from the most dangerous vector production room. It is that you are. Safety is ensured by combining these three measures. Further, there is an advantage that the productivity is improved because there is no waste in the flow of goods and people, the quality is kept constant because the quality control is easy, and the yield is improved.

Regarding the first measure, which is the regulation of the flow lines of workers, raw materials, instruments, vectors, etc., the workers, raw materials, instruments, vectors, etc. enter through the doors on the entrance side of each room and the exit side. The flow of people and objects is automatically regulated in one direction by protecting the movement of moving out of the door to the next room. In this way, the flow line is in one direction, with the workers before the vector production and the raw materials and equipment necessary for vector production,
Since there is no contact with the workers who have created the vector or the contaminated wastes and the vector that is the product, products such as dangerous vectors and wastes from the vector production room are upstream from the process. There is no fear of moving to the changing room. Further, in the direction of the undressing room, which is the downstream side of the process from the vector production room, contaminants such as waste are configured to go out after being sterilized in the sterilization room. There is no risk of dangerous contaminants leaking out.

The above-mentioned second and third countermeasures will be described with reference to FIGS. 3A to 3C and 4A to 4C.

FIGS. 3A to 3C are diagrams showing the internal pressure of each room and the flow of air between the rooms in the vector production facility according to the embodiment of the present invention shown in FIG. In FIGS. 3A to 3C, the relationship between the room pressures of the chambers and the air flow between the chambers are indicated by arrows. In all of FIGS. 3A, 3B, and 3C, the air pressure in the vector production chamber is less than atmospheric pressure, and the chambers adjacent to each other between the vector production chamber and the changing room and between the vector production room and the undressing chamber are adjacent to each other. It is shown that there is at least one of the chamber for sucking air and discharging it to the outside of the chamber and the chamber for sucking air from the outside and blowing air to the adjacent chamber. Therefore, the air in the vector production room does not flow into the changing room or the undressing room.

For example, in the case of FIG. 3A, the room pressure of the first pre-airlock room between the vector preparation room and the changing room is lower than that of the adjacent preparation room and vector preparation room. Therefore, the air in the preparation room and the vector production room is
It is sucked into the air lock front chamber and is discharged to the outside through the HEPA filter. Therefore, the first pre-airlock chamber functions as a barrier that blocks the flow of air from the vector preparation chamber to the preparation chamber. Similarly, since the airlock rear chamber also has a lower air pressure than the adjacent vector preparation chamber and sterilization chamber, it also functions as a barrier that blocks the flow of air from the vector preparation chamber to the sterilization chamber.

In the case of FIG. 3B, the air pressures of the first air-lock front chamber and the first air-lock rear chamber are set higher than the air pressures of the two adjacent chambers. In this case, the air sucked from the outside only flows from the first pre-airlock chamber or the post-airlock chamber to the adjacent chambers on both sides, and the air in the vector preparation chamber is changed to the pre-first airlock chamber. It does not flow past the chamber to the preparation chamber or after the airlock to the chamber and into the sterilization chamber. Therefore, in this case as well, the first airlock front chamber and the first airlock rear chamber function as barriers that block the flow of air between the chambers on both sides thereof.

As described above, it is understood that the first airlock front chamber and the first airlock rear chamber exhibit the function as a barrier regardless of whether the room pressure is higher or lower than the adjacent chambers on both sides. Therefore, as shown in FIG. 3C,
In the same vector production facility, a first pre-airlock chamber whose room pressure is higher than both chambers and a first post-airlock chamber whose room pressure is lower than both chambers, or vice versa. It does not matter even if the airlock and the rear room are mixed.

FIGS. 4A to 4C are diagrams showing the internal pressure of each room and the flow of air between the rooms in the vector production facility according to the embodiment of the present invention shown in FIG. In this case, the preparation room 3 shown in FIG. 1 is divided into a first preparation room 3-1 and a second preparation room 3-2, and the first preparation room 3-1 and the second preparation room 3-1. A second airlock front chamber 5 is provided between the chamber 3-2 and the chamber 3-2.

FIG. 4A shows that when the room pressures of the first airlock front chamber, the second airlock front chamber and the airlock rear chamber are all lower than the room pressures on both sides, FIG.
When the room pressures of the chambers are both higher than the room pressures on both sides, FIG. 4C shows that these three chambers have a room pressure higher than the chambers on both sides (the first pre-airlock chamber in FIG. 4C) and The case where the chambers whose pressure is lower than the chambers on both sides (the second pre-airlock chamber and the second post-airlock chamber in FIG. 4C) coexist is shown. As described above, these three chambers each function as a barrier.

When a changing room and a changing room are provided, the room pressure of these rooms may be atmospheric pressure, but positive pressure is preferable. As is clear from the air flows shown in FIGS. 3A to 3C and 4A to 4C, the room pressure in the changing room and the changing room is set to a positive pressure higher than that in the preparation room, the first preparation room, or the sterilization room. This is because by setting it, the function as a barrier can be exhibited in the changing room and the changing room. Further, it is desirable that the sterilization chamber has a negative pressure lower than atmospheric pressure.

The adjustment of the air pressure in each chamber as shown in FIGS. 3A to 3C and FIGS. 4A to 4C can be performed in accordance with the relationship between the amount of air supplied to each chamber and the amount of exhaust air from each chamber. it can. A sirocco fan, a propeller-type fan, or the like can be used as the air supply / exhaust device. The indoor pressure adjusting means is an air flow rate adjusting means such as a damper attached to a part of a duct connecting these fans and the air supply port or the exhaust port provided in each room with the fan, and the indoor pressure adjusting means. It may be composed of a detector or the like. Although the air supply port or the exhaust port is preferably provided in all the rooms, it is not always necessary to provide them in all the rooms. As shown in FIG. 3A, FIG. 3C and FIG. 4A, FIG. 4C, when the preparation chamber, the first preparation chamber, the second preparation chamber, the sterilization chamber, etc. have neither an air supply port nor an exhaust port. However, it is possible to control the flow of air.

The chamber internal pressure of each chamber when performing the vector preparation work is such that the air flow is as shown in FIG. 3A to FIG. 3C or FIG. 4A to FIG.
As shown in C, the room pressure may be such that it does not flow outside from the vector preparation room. Therefore, it is not necessary to strictly control the absolute value of the indoor pressure of each room. However, when considering the balance of the air pressure of the entire vector production facility, the ease of controlling the indoor pressure, the workability, etc., Table 1 shows examples of preferable indoor pressure and preferable range (unit: Pa) of each room. is there.

[0058]

[Table 1]

The air lock chambers such as the first air lock front chamber, the second air lock front chamber and the air lock rear chamber are
It must act to block the flow of air between the chambers on both sides of the air lock chamber.
Therefore, the room pressure of the airlock chamber is as shown in FIGS. 3A to 3C.
And as shown in FIGS. 4A-4C, it must be either higher or lower than the chambers on either side. In that case, the difference in the room pressure is preferably 15 Pa or more.

The chamber pressures in the preparation chambers other than the air lock chamber, the first and second preparation chambers, the sterilization chamber, etc. are as shown in FIGS.
As described with reference to C or FIGS. 4A to 4C, an air pressure capable of controlling the air flow between the chambers may be selected. That is, it is preferable to select a pressure suitable for each vector production facility with reference to the indoor pressure of the vector production room, airlock room, changing room, undressing room and the like. In addition, the air pressure inside the safety cabinet must be kept lower than the outside pressure of the safety cabinet, that is, the room pressure inside the vector production chamber, as a matter of course.

It is desirable that the air in each room is clean, and in accordance with JIS B 9920 (1989), the class 4 is provided between the preparation room or the second preparation room and the room after the air lock.
The degree of cleanliness is about 5 for the changing rooms, the first preparation room, the sterilization room, the undressing room, etc. For other rooms, the quality control room is for class 4, the vector storage room and the control room are for class 5 It is desirable to maintain.

In order to maintain the cleanliness of the air, the structure and air supply system of a normally used clean room may be adopted. In the case of the vector production facility according to the present invention, the structure of the clean room may be, for example, a non-laminar flow type or a turbulent flow type. In addition, a clean air is supplied to the room by using a filter such as a HEPA filter for the air supply system.

A HEPA filter is installed in the exhaust system so as to prevent contaminated suspended solids and dangerous suspended solids from leaking out of the facility. In particular, it is necessary to reliably trap the air discharged from the vector production chamber. Therefore, a HEPA filter is used for the exhaust system of the vector production room and the safety cabinet installed in the vector production room.

In the case of the vector production facility according to the present invention, rather than circulating air between the exhaust system and the air supply system, the air in the exhaust system is discharged outside the facility and new air is supplied to the air supply system. It is advisable to use a total ventilation system that supplies The total ventilation system is desirable because it prevents any contaminated or dangerous substances floating in the facility air from returning to the facility.

When carrying out the work of producing the vector, the worker changes into a work clothes such as dust-free clothes which does not easily generate dust in the changing room. After that, the preparation room and the first airlock front room,
Alternatively, the vector preparation chamber is entered according to the flow line of the first preparation chamber, the second pre-airlock chamber, the second preparatory chamber and the first pre-airlock chamber. During this period, the raw materials, tools, etc. necessary for vector production are prepared. An interlock system is adopted so that the door on the entrance side and the door on the exit side do not open at the same time when passing through the first and second airlock front chambers.

When a vector is prepared using a virus in the vector preparation room, the work is always carried out in a safety cabinet. Even if a virus is not used, it is better to work in a safety cabinet when producing a gene recombinant. In the vector production room, the produced vector is cultured and propagated.
Therefore, in the vector production room, a lot of wastes such as dangerous products and disposable containers used for production and contaminated are generated. In addition, there is a possibility that dangerous substances may adhere to work clothes worn by the worker.

These dangerous substances must be moved toward the downstream side of the process, that is, the sterilization chamber side. That is, it moves from the vector preparation room to the sterilization room through the airlock post room. At this time, the door in the rear room of the airlock is opened and closed by an interlock system so that the door on the entrance side and the door on the exit side cannot be opened at the same time. In the sterilization room, take measures to prevent contaminated waste, etc., from leaking outside. Waste and potentially contaminated material should be passed through the autoclave and out of the sterilization room.

When the prepared vector is transferred to the adjacent vector storage room, it is preferable to use the product transfer port provided in a part of the partition wall. The product delivery port is preferably in the shape of a box having an opening / closing door on the side of the sterilization chamber and the side of the vector storage chamber, and has a structure capable of keeping the inside airtight. By using such a product delivery port, an operator can safely and easily transport a vector, which is a product, to the vector storage chamber without passing through a preparation room or the like.

The worker goes through the sterilization chamber to the dressing room, and changes clothes there. The work clothes used should be sterilized separately.

By adopting such a vector production method, it is possible to efficiently produce a vector while sufficiently ensuring safety.

[0071]

EFFECTS OF THE INVENTION According to the vector production facility or the vector production method of the present invention, the flow lines of the operator, the raw materials, and the tools necessary for the vector production are limited in one direction. There is no contact with subsequent workers. In addition, there is no risk that the air in the vector production room leaks to the outside, and all wastes and exports can be sterilized by a high-pressure steam sterilizer before being exported to the outside. As described above, since contaminated air and contaminants do not leak outside from the vector production facility, the physical containment level P2 or P3 and GMP specified in the recombinant DNA experimental guideline are defined. It is advantageous in fully satisfying the safety standards regarding containment.
In addition, since the flow line in the facility is limited to one direction, there is no waste in the flow of goods and people, the quality control is easy because the quality control is easy and the yield is improved. Since it can be placed, it has the advantage of being excellent in productivity.

[Brief description of drawings]

FIG. 1 is a plan view showing an arrangement example of each room in a vector production facility according to an embodiment of the present invention.

FIG. 2 is a plan view showing an arrangement example of each room in a vector production facility according to another embodiment of the present invention.

FIG. 3A is a diagram showing an aspect of the indoor pressure of each room and the flow of air between the rooms in the vector production facility according to the embodiment of the present invention shown in FIG. 1.

FIG. 3B is a diagram showing another aspect of the indoor pressure of each room and the flow of air between the rooms in the vector production facility according to the embodiment of the present invention shown in FIG. 1.

FIG. 3C is a diagram showing still another aspect of the indoor pressure of each room and the flow of air between the rooms in the vector production facility according to the embodiment of the present invention shown in FIG. 1.

FIG. 4A is a diagram showing an aspect of the indoor pressure of each room and the flow of air between the rooms in the vector production facility according to another embodiment of the present invention shown in FIG.

FIG. 4B is a diagram showing another mode of the indoor pressure of each room and the flow of air between the rooms in the vector production facility according to another embodiment of the present invention shown in FIG.

FIG. 4C is a diagram showing still another aspect of the indoor pressure of each room and the flow of air between the rooms in the vector production facility according to another embodiment of the present invention shown in FIG.

[Explanation of symbols]

2 Vector production room 3 preparation room 3-1 First preparation room 3-2 Second preparation room 5 First Airlock Front Room 6 Second Airlock Front Room 7 Airlock rear room 8 Sterilization room 9 High-pressure steam sterilizer 10 changing rooms 11 dressing room 15 safety cabinet

Continued front page    (72) Inventor Sachiichi Kuda             38-31 Suehiro-cho, Kadoma-shi, Osaka 601 Mona Lisa F-term (reference) 4B024 AA19 CA01 CA09 EA02 EA04                       HA17 HA20                 4B029 AA23 BB20

Claims (11)

[Claims] 1. A preparation chamber, a first pre-airlock chamber, a vector preparation chamber, a post-airlock chamber, and a sterilization chamber, which allow an operator to enter the preparatory chamber and then the first pre-airlock chamber,
A door is provided between the chambers to move the chambers in the order of the vector preparation chamber and the airlock rear chamber, and to form a restricted flow line of going out through the sterilization chamber. Furthermore, a vector production facility equipped with indoor pressure control means for controlling the indoor pressure of each room. 2. The preparatory chamber comprises a first preparatory chamber, a second preparatory chamber and a second pre-airlock chamber provided between the first preparatory chamber and the second preparatory chamber. , The worker is the first
2. The door is provided between the preparation chambers, the second pre-airlock chamber, and the second preparation chamber so as to form a restricted flow line that moves in that order. Vector production facility. 3. The vector production facility according to claim 1, further comprising a changing room outside the entrance door of the preparation room. 4. The vector production facility according to claim 2, wherein a changing room is provided outside the entrance door of the first preparation room. 5. The vector preparation facility according to claim 1, further comprising a dressing room outside the exit door of the sterilization processing room. 6. The room pressure of the first pre-airlock chamber is lower or higher than that of the preparation chamber and the vector preparation chamber, and the rear airlock chamber is lower than that of the vector preparation chamber and the sterilization chamber. 6. The vector production facility according to claim 1, 3 or 5, wherein the vector production chamber and the sterilization chamber are maintained at a pressure higher than atmospheric pressure or higher than atmospheric pressure. 7. The room pressure in the second pre-airlock chamber is lower or higher than that in the first preparatory chamber and the second preparatory chamber, and the first pre-airlock chamber is in the second preparatory chamber. Lower or higher than the preparation room and the vector preparation room, the airlock post-chamber is lower or higher than the vector preparation room and the sterilization room, and the vector preparation room and the sterilization room are lower than atmospheric pressure. The vector production facility according to claim 2, 4 or 5, which is maintained. 8. The vector storage room according to claim 1, 3, 5 or 6 provided with a vector storage room in which an installation position and a door position are determined so that an operator can enter and exit without going through the preparation room. Vector production facility. 9. A quality control room in which an installation position and a position of a door are determined so that an operator can enter and exit without going through the preparation room, according to claim 1, 3, 5, 6 or 8. The described vector production facility. 10. A preparation room, a first pre-airlock room, a vector preparation room, a post-airlock room, and a sterilization room, where an operator prepares a preparatory room, a first pre-airlock room, a vector preparation room, and Doors are provided between the chambers so as to form a restricted flow line of moving through the chambers in the order of the airlock and then exiting through the sterilization chamber. With the vector production facility equipped with the indoor pressure adjusting means for adjusting the pressure, the indoor pressure is lower or higher than the preparatory chamber and the vector production chamber, and the airlock rear chamber is A method for producing a vector, which is lower or higher than the vector production chamber and the sterilization chamber, and is maintained at a pressure lower than the atmospheric pressure in the vector production chamber and the sterilization chamber. 11. A first preparation chamber, a second pre-airlock chamber,
A second preparation room, a first pre-airlock room, a vector preparation room, a post-airlock room and a sterilization processing room are provided, and an operator prepares a first preparation room, a second pre-airlock room, and a second pre-airlock room. In order to form a restricted flow line of moving each chamber in the order of the preparation chamber, the first pre-airlock chamber, the vector preparation chamber and the post-airlock chamber, and going out through the sterilization treatment chamber,
A door is provided between the rooms, and the indoor pressure is adjusted by the vector preparation facility equipped with an indoor pressure adjusting means for adjusting the indoor pressure of each room. Lower or higher than the chamber and said second preparation chamber,
The first pre-airlock chamber is lower or higher than the second preparatory chamber and the vector preparation chamber, the rear airlock chamber is lower or higher than the vector preparation chamber and the sterilization chamber, and A method for producing a vector, in which the vector production room and the sterilization chamber are maintained at a pressure lower than atmospheric pressure.