Disposable bioreactor culture bag for virus packaging

Abstract

The utility model provides a disposable bioreactor culture bag for virus packaging, which comprises a bag body, an isolating layer is arranged in the bag body, the upper part of the isolating layer is an upper layer, and the lower part of the isolating layer is a lower layer; the top of the bag body is provided with a first liquid inlet pipeline and a second liquid inlet pipeline which extend into the upper layer; a stirrer and a paddle are arranged on the lower layer in the bag body; the isolation layer is further provided with a connecting port, and the upper layer and the lower layer in the bag body are communicated through the connecting port. According to the utility model, the connecting port is arranged, so that a virus packaging compound can be quickly added into a lower-layer cell culture solution, the advantages of instantaneous transduction and difficulty in bubbling are reflected, the mode of dripping test transduction during virus packaging of an existing disposable reaction bag can be replaced, and the virus packaging and releasing efficiency is improved.

Description

Technical Field

[0001] The utility model provides a disposable bioreactor culture bag for virus packaging, belonging to the field of biotechnology. Background Art

[0002] Transient transfection is one of the ways to introduce DNA into eukaryotic cells. Transient transfection of mammalian cells can quickly express highly active proteins in a short period of time and is widely used, especially in viral vectors such as adeno-associated virus (AAV) and lentivirus (LV). By performing a large number of transient transfections on HEK293 cells that are adapted to suspension culture, a large number of viral vectors can be obtained.

[0003] Biotechnology requires solutions that can support large-scale cell culture growth. Disposable bioreactors are not only systems that can successfully support large-scale cell culture, but also reduce the risk of cross-contamination between viral vectors.

[0004] The existing disposable bioreactor culture bag has an integrated bag body and a bottom sealing structure, which is sealed and connected to the bag body. The inner rotor is pivotally connected to the sealing structure, and the inner rotor is located inside the bag body. The inner rotor is provided with a stirring blade. The sealing structure is provided with an outer rotor matching groove, which is located outside the bag body and can match the outer rotor, and the outer rotor is connected to the motor. The disposable bioreactor culture bag has the following disadvantages: 1. During the viral vector packaging process, when operating in large volumes, the transfection complex depends on the pipeline originally prepared in the disposable reaction bag, resulting in a longer operation time for introducing the complex due to different pipelines, which also creates challenges in batch difference control and affects the transient transfection efficiency; 2. During the cell culture process, as the cells grow, there will usually be a certain volume of foam on the liquid surface. It is easy to foam when adding liquid directly from the liquid surface. The complex remains in the upper foam and cannot be immediately added to the cell culture medium, which affects the efficiency of virus packaging and release.

[0005] Therefore, it is necessary to design and modify the disposable reactor culture bags used in the virus packaging process to effectively improve the AAV packaging efficiency and yield. Utility Model Content

[0006] In order to solve the problems raised in the above background technology, the purpose of the utility model is to provide a disposable bioreactor culture bag for viral vector packaging, which has the advantages of instantaneous transfection and solves the problem of easy foaming during dripping in the existing virus packaging process.

[0007] In order to achieve the above purpose, the utility model provides the following technical solutions:

[0008] A disposable bioreactor culture bag for virus packaging comprises a bag body, wherein an isolation layer is arranged inside the bag body, the upper layer is above the isolation layer, and the lower layer is below the isolation layer; a first liquid inlet pipeline and a second liquid inlet pipeline are arranged on the top of the bag body and extend into the upper layer; an agitator and a paddle are arranged in the lower layer inside the bag body; and a connecting port is also arranged on the isolation layer, through which the upper layer and the lower layer in the bag body are connected.

[0009] The connecting port is cylindrical, the bottom opening edge of the connecting port is fixed on the surface of the isolation layer, and the connection between the isolation layer and the bottom opening edge of the connecting port is an easily-tearable connection. The top opening of the connecting port is arranged on the bag wall of the bag body. A handle is also arranged in the connecting port. The lower end of the handle is fixed in the area where the isolation layer is located in the bottom opening of the connecting port. The handle extends from the top opening of the connecting port to the outside of the bag body. When the handle is pulled, the isolation layer located in the bottom opening area of ​​the connecting port is pulled by the handle and torn along the easily-tearable connection to connect the upper and lower layers in the bag body.

[0010] Compared with the prior art, the beneficial effects of the utility model are as follows:

[0011] The utility model can quickly add the virus packaging complex to the lower cell culture fluid by providing a connection port, embodying the advantages of instantaneous transduction and not prone to foaming, and can replace the existing method of dripping test transduction during virus packaging in disposable reaction bags, thereby improving the virus packaging and release efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 It is a front view schematic diagram of the structure of the utility model;

[0013] Figure 2 It is a partially enlarged schematic diagram of the structure of the utility model. DETAILED DESCRIPTION

[0014] The specific technical solution of the utility model is explained in conjunction with the accompanying drawings.

[0015] like Figure 1 and Figure 2 As shown, a disposable bioreactor culture bag for virus packaging comprises a bag body 2, an isolation layer 1 is arranged inside the bag body 2, the upper part of the isolation layer 1 is an upper layer, and the lower part is a lower layer; a first liquid inlet pipeline 7 and a second liquid inlet pipeline 8 are arranged on the top of the bag body 2 and extend into the upper layer; a stirrer 5 and a paddle 6 are arranged on the lower layer inside the bag body 2;

[0016] The bag bag also includes a connecting port 4, which is cylindrical, and the bottom opening edge of the connecting port 4 is fixed on the surface of the isolation layer 1, and the connection between the isolation layer 1 and the bottom opening edge of the connecting port 4 is an easily-tearable connection 9. The top opening of the connecting port 4 is arranged on the bag wall of the bag body 2. A handle 3 is also arranged in the connecting port 4. The lower end of the handle 3 is fixed in the area where the isolation layer 1 is located in the bottom opening of the connecting port 4. The handle 3 extends from the top opening of the connecting port 4 to the outside of the bag body 2. When the handle 3 is pulled, the handle 3 pulls the isolation layer 1 located in the bottom opening area of ​​the connecting port 4, and is torn along the easily-tearable connection 9 to connect the upper layer and the lower layer in the bag body 2.

[0017] The utility model first connects two external mixed material containers with the first liquid inlet pipeline 7 and the second liquid inlet pipeline 8 on the top of the bag body 2, and simultaneously adds the mixed solutions into the upper space of the bag body 2. After the compound is left to stand for a long time, the external handle 3 is held to tear open the bottom of the connection port 4 of the isolation layer 1, and the virus packaging compound can quickly flow along the bag wall into the cell culture solution in the lower layer. Finally, the agitator 5 is used to quickly stir the compound until it is uniform, thereby facilitating rapid virus packaging and transduction, replacing the existing method of dripping and testing transduction during virus packaging in disposable reaction bags, and improving the efficiency of virus packaging and release.

Claims

1. A disposable bioreactor culture bag for virus packaging, characterized in that: The bag body (2) comprises a bag body (2), wherein an isolation layer (1) is provided inside the bag body (2), the upper layer is above the isolation layer (1), and the lower layer is below the isolation layer (1); a first liquid inlet pipeline (7) and a second liquid inlet pipeline (8) are provided at the top of the bag body (2) and extend into the upper layer; an agitator (5) and a paddle (6) are provided in the lower layer inside the bag body (2); and a connecting port (4) is also provided on the isolation layer (1), and the upper layer and the lower layer in the bag body (2) are connected via the connecting port (4).

2. A disposable bioreactor culture bag for virus packaging according to claim 1, characterized in that: The connection port (4) is cylindrical, the bottom opening edge of the connection port (4) is fixed on the surface of the isolation layer (1), and the connection between the isolation layer (1) and the bottom opening edge of the connection port (4) is an easily tearable connection (9), and the top opening of the connection port (4) is arranged on the bag wall of the bag body (2).

3. A disposable bioreactor culture bag for virus packaging according to claim 2, characterized in that: The connection port (4) is also provided with a handle (3), the lower end of the handle (3) is fixed in the area of ​​the isolation layer (1) located at the bottom opening of the connection port (4), and the handle (3) extends from the top opening of the connection port (4) to the outside of the bag body (2). When the handle (3) is pulled, the handle (3) pulls the isolation layer (1) located in the bottom opening area of ​​the connection port (4), and is torn along the tearable connection (9), thereby connecting the upper layer and the lower layer in the bag body (2).

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