Microcarrier crosslinking cleaning system

Abstract

According to the microcarrier cross-linking cleaning system provided by the invention, the microcarrier is added into the filter, so that the microcarrier can be conveniently separated from a cross-linking reagent, a cleaning solution and other reagents, and the cross-linking of the microcarrier is prevented from being interfered by a production reagent; the microcarrier can suspend in the cross-linking agent through the stirring of the stirring device, so that the cross-linking uniformity of the microcarrier is ensured; the cross-linked microcarrier can suspend in the cleaning liquid through stirring of the stirring device, residues of a cross-linking agent in the cross-linked microcarrier are reduced, and the residues are within a controllable range. In the system, the stirring device is arranged in the tank body instead of the filter, so that the physical stability of the microcarrier before crosslinking is ensured, the microcarrier is ensured not to be in contact with the stirring device, the microcarrier is further prevented from being broken, and the quality of the microcarrier is improved. The system can continuously realize continuous production of micro-carrier pretreatment, micro-carrier cross-linking, cross-linked micro-carrier cleaning and collection.

Description

Technical Field

[0001] The utility model belongs to the technical field of cell culture and relates to a microcarrier cross-linking cleaning system. Background Art

[0002] Microcarrier culture is an emerging large-scale cell culture technology. It uses tiny particles called microcarriers as cell carriers. Cells are suspended in a culture medium through agitation, allowing them to multiply into a monolayer on or within the carriers. Microcarrier culture is widely used in the production of vaccines and genetically engineered products. It is currently recognized as one of the most promising large-scale animal cell culture technologies, combining the advantages of both suspension and adherent cultures while being easily scaled up.

[0003] The stability and mechanical property strength of uncrosslinked microcarrier are low, can not be directly applied, therefore, conventionally use crosslinked method to carry out modification, to enhance its physical property. Commonly used crosslinking method is physical crosslinking method and chemical crosslinking method, but the crosslinking degree of physical crosslinking method is lower and inhomogeneous, therefore, conventional crosslinking method is chemical crosslinking method. Chemical crosslinking agent commonly used at present has glutaraldehyde, formaldehyde, ethylene glycol, genistein etc., these crosslinking agents can have cytotoxicity conventionally, cause the survival rate of normal cells to reduce, therefore, through chemical crosslinked microcarrier need through fully cleaning, to control the residual amount of crosslinking agent, and then for cell culture.

[0004] During the microcarrier preparation, cross-linking, and cleaning processes, existing equipment is usually small-scale equipment at the experimental stage, which cannot achieve continuous production, and the prepared microcarriers are not uniform. Utility Model Content

[0005] The purpose of the utility model is to provide a microcarrier cross-linking cleaning system to solve the problem that the existing device cannot achieve continuous production.

[0006] In order to achieve the above purpose, the present invention adopts the following technical solutions:

[0007] The utility model provides a microcarrier cross-linking cleaning system, comprising a bracket and a fixing frame located above the bracket, a motor is arranged inside the bracket, a tank body is fixedly arranged on the fixing frame, a stirring device is arranged inside the tank body, and the output end of the motor is connected to the stirring device; a liquid outlet pipe is provided at the bottom of the tank body, a sealing cover is provided at the top of the tank body, and a connected feed port and a filter are provided on opposite sides of the sealing cover, and the filter is located inside the tank body and above the stirring device.

[0008] Preferably, the filter is a funnel-shaped filter.

[0009] Preferably, the filter includes a filter screen and a filter membrane limiter located inside, and a filter membrane is placed inside the filter membrane limiter; a limit plate and a slot are provided on the top of the filter screen, the limit plate is detachably arranged on the sealing cover, and the filter membrane limiter is rotatably engaged in the slot.

[0010] Preferably, the stirring device includes a stirring shaft and stirring blades connected to each other, and the stirring shaft is connected to the output end of the motor.

[0011] Preferably, the sealing cover is provided with at least one pair of the feed port and the filter.

[0012] Preferably, a liquid inlet is further provided on the top of the sealing cover, and the liquid inlet is connected to the interior of the tank body.

[0013] Preferably, a liquid outlet valve is provided on the liquid outlet pipe.

[0014] Preferably, a support column is provided between the bracket and the fixing frame.

[0015] Preferably, the system further comprises a shell, wherein the shell is located outside the bracket and the tank body, and the sealing cover and the feed port are exposed at the top of the shell.

[0016] Preferably, the liquid outlet valve on the liquid outlet pipe is exposed outside the housing.

[0017] The utility model has the following beneficial effects:

[0018] (1) By adding microcarriers into the filter, the separation of microcarriers from crosslinking reagents, cleaning solutions and other reagents can be facilitated, so that the crosslinking of microcarriers is not interfered with by production reagents.

[0019] (2) The pore size of the filter membrane is smaller than the standard diameter of the microcarrier, so as to facilitate the filtration of microcarrier fragments with a diameter smaller than the standard diameter of the microcarrier and ensure the uniformity of the microcarrier.

[0020] (3) The stirring of the stirring device can suspend the microcarriers in the crosslinking agent, ensuring the uniformity of the crosslinking of the microcarriers; at the same time, the stirring of the stirring device can suspend the crosslinked microcarriers in the cleaning solution, reducing the residue of the crosslinking agent in the crosslinked microcarriers and keeping the residue within a controllable range.

[0021] (4) The stirring device is set inside the tank instead of inside the filter, which ensures the physical stability of the microcarriers before cross-linking. At the same time, it ensures that the microcarriers will not come into contact with the stirring device, and thus the microcarriers will not be broken, thereby improving the quality of the microcarriers.

[0022] (5) Both the feed port and the liquid inlet are configured in a funnel shape to facilitate the entry of the microcarriers and the crosslinking agent into the filter; the filter is configured in a funnel shape to facilitate the sedimentation and collection of the microcarriers.

[0023] (6) The microcarrier cross-linking and cleaning system can continuously realize the continuous production of microcarrier pretreatment, microcarrier cross-linking, cross-linked microcarrier cleaning and collection. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 A schematic diagram of the three-dimensional structure of the microcarrier cross-linking cleaning system provided in an embodiment of the present application;

[0025] Figure 2 A cross-sectional view of a microcarrier cross-linking cleaning system provided in an embodiment of the present application;

[0026] Figure 3 A schematic diagram of the three-dimensional structure of the filter provided in an embodiment of the present application;

[0027] Symbols represent:

[0028] 1- bracket, 2- fixed frame, 3- motor, 4- tank body, 5- stirring device, 6- liquid outlet pipe, 7- sealing cover, 8- feed port, 9- filter, 10- liquid outlet valve, 11- liquid inlet, 12- support column, 13- shell;

[0029] 501- stirring shaft, 502- stirring blade;

[0030] 901-filter screen, 902-filter membrane limiter, 903-limiting plate, 904-card slot. DETAILED DESCRIPTION

[0031] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0032] The present invention provides a microcarrier cross-linking cleaning system, which includes a bracket 1, a fixing frame 2, a tank 4, a motor 3 and other components. Figure 1 、 2 The bracket 1 is located at the bottom and is used to support the fixing frame 2, the tank body 4, etc. A motor 3 is provided inside the bracket 1, and the motor 3 is used to drive the stirring device 5 inside the tank body 4 to stir.

[0033] The fixing frame 2 is located above the bracket 1 and is arranged on the top of the bracket 1 through a support column 12, and the top of the fixing frame 2 is provided with a tank body 4. In the embodiment of the present application, the support column 12 is a shock-absorbing support column to prevent the tank body 4 from shaking during the stirring process and improve the stability of the system.

[0034] The top of the tank body 4 is provided with a sealing cover 7, which is used to seal the top of the tank body 4 to prevent the liquid inside the tank body 4 from splashing out. A feed port 8 and a filter 9 are provided on opposite sides of the sealing cover 7. The filter 9 is located inside the tank body 4, and the feed port 8 is located outside the tank body 4. Microcarriers enter the filter 9 through the feed port 8. The filtering action of the filter 9 removes microcarriers with diameters smaller than the standard diameter, microcarrier fragments, and liquids, and also facilitates the recovery of cleaned cross-linked microcarriers. Furthermore, to increase the filtration capacity of the microcarriers, the sealing cover 7 is provided with at least one pair of feed ports 8 and filters 9.

[0035] The filter 9 in the embodiment of the present application is a funnel-shaped filter, and the filter 9 is detachably connected to the sealing cover 7. Figure 3 As shown. The filter 9 in the embodiment of the present application includes a filter screen 901, a filter membrane stopper 902 and a filter membrane (not shown in the figure). A stopper plate 903 and a card slot 904 are provided on the top of the filter screen 901. The stopper plate 903 is detachably arranged on the sealing cover 7 by a rotational card connection, thereby realizing a detachable connection between the filter 9 and the sealing cover 7. A filter membrane stopper 902 is provided inside the filter screen 901, on which a filter membrane is placed and fixed. The filter screen 901 is clamped in the card slot 904 by a rotational card connection, thereby realizing the fixation of the filter membrane and the fixation between the filter screen 901 and the filter membrane stopper 902.

[0036] In the embodiment of the present application, the pore size of the filter membrane is smaller than the standard diameter of the microcarrier, so as to facilitate filtering microcarriers and fragments with diameters smaller than the standard diameter of the microcarrier, thereby ensuring the uniformity of the microcarrier.

[0037] In addition, a stirring device 5 is provided inside the tank body 4, and the stirring device 5 is connected to the output end of the motor 3. Thus, when the motor 3 is running, it can drive the stirring device 5 to stir. Specifically, the stirring device 5 includes a stirring shaft 501 and a stirring blade 502 connected to each other, and the stirring shaft 501 is connected to the output end of the motor 3. When the motor 3 is running, the stirring shaft 501 rotates and drives the stirring blade 502 to rotate, thereby stirring the liquid inside the tank body 4 to form a vortex flow field. The stirring of the vortex flow field inside the tank body 4 drives the liquid and microcarriers in the filter 9 to stir.

[0038] To prevent the stirring device 5 from damaging the filter 9 during stirring, the stirring device 5 is located below the filter 9. Since the stirring device 5 is located inside the tank 4 rather than inside the filter 9, the stirring of the stirring device 5 does not break the microcarriers, thereby improving the quality of the microcarriers.

[0039] A liquid outlet pipe 6 is provided at the bottom of the tank body 4 for discharging microcarrier fragments, liquid, etc. In order to facilitate the discharge of various liquids from the tank body 4 , a liquid outlet valve 10 is provided at the outlet of the liquid outlet pipe 6 .

[0040] In addition, a liquid inlet 11 is provided at the top of the sealing cap 7, which communicates with the interior of the tank body 4. The liquid inlet 11 is used to introduce a crosslinking agent liquid or a cleaning solution into the tank body 4 to facilitate crosslinking with the microcarriers or to clean the crosslinked microcarriers. In the embodiment of the present application, both the feed inlet 8 and the liquid inlet 11 are provided with sealing caps to prevent dust and other contaminants from entering the interior of the tank body 4 and the filter 9. Furthermore, to facilitate the addition of materials to the tank body 4 and the filter 9, the feed inlet 8 and the liquid inlet 11 are both funnel-shaped.

[0041] In this embodiment of the present application, the microcarrier crosslinking and cleaning system further includes a housing 13, which is located outside the support 1 and tank 4 to protect them. The sealing cap 7, feed port 8, and liquid inlet 11 are all exposed at the top of the housing 13, facilitating the addition of material to the tank 4 and filter 9. The outlet valve 10 on the liquid outlet pipe 6 is exposed at the housing 13 to facilitate the collection of waste liquid.

[0042] The microcarrier crosslinking and cleaning system provided in the embodiments of the present application is used for continuous preparation of microcarriers, including microcarrier pretreatment, microcarrier crosslinking, crosslinked microcarrier cleaning and collection. The specific process is as follows:

[0043] (1) Microcarrier pretreatment

[0044] Open the sealing cap and liquid outlet valve 10 at the top of the feed port 8, and allow the prepared microcarrier suspension to enter the filter 9 through the feed port 8. During this process, the filter 9 filters the microcarriers, retaining microcarriers of standard diameter inside the filter 9, while microcarriers smaller than the standard diameter, microcarrier fragments, and liquid are discharged through the liquid outlet pipe 6.

[0045] (2) Microcarrier cross-linking

[0046] Close the liquid outlet valve 10, open the sealing cover on the top of the liquid inlet 11, and add the crosslinking agent liquid to the tank body 4. After the crosslinking agent liquid submerges the filter 9, close the sealing cover on the top of the liquid inlet. Turn on the switch of the motor 3 so that the motor 3 drives the stirring shaft 501 and the stirring blade 502 to rotate, thereby stirring the liquid inside the tank body 4 and forming a vortex flow. The vortex flow drives the crosslinking agent and microcarriers inside the filter 9 to rotate, so that the microcarriers are in a state of motion, ensuring uniform contact between all microcarriers and the crosslinking agent, and ensuring the uniformity of the crosslinking of the microcarriers. After crosslinking is completed, turn off the motor 3 switch, the stirring ends, and the crosslinked microcarriers are settled inside the funnel-shaped filter 9. Open the liquid outlet valve 10, and the crosslinking agent is discharged through the liquid outlet pipe 6.

[0047] (3) Cleaning and collection of cross-linked microcarriers

[0048] Close the liquid outlet valve 10, open the sealing cover on the top of the liquid inlet 11, and add cleaning liquid to the inside of the tank body 4. When the cleaning liquid submerges the filter 9, close the sealing cover on the top of the liquid inlet 11. Turn on the switch of the motor 3 so that the motor 3 drives the stirring shaft 501 and the stirring blade 502 to rotate, thereby stirring the liquid inside the tank body 4 and forming a vortex flow. The vortex flow drives the cleaning liquid and cross-linked microcarriers inside the filter 9 to rotate, so that the cross-linked microcarriers are in a state of motion, ensuring that all cross-linked microcarriers are in uniform contact with the cleaning liquid and reducing the residue of the cross-linking agent on the cross-linked microcarriers. After the cleaning is completed, turn off the switch of the motor 3, the stirring is completed, and the cross-linked microcarriers are settled inside the funnel-shaped filter 9. Open the liquid outlet valve 10, and the cleaning liquid is discharged through the liquid outlet pipe 6. The cleaning process can be repeated many times until the residual amount of cross-linking agent in the cleaning liquid discharged from the liquid outlet pipe 6 meets the requirements.

[0049] When the residual amount of cross-linking agent in the cleaning liquid discharged from the liquid outlet pipe 6 meets the requirements, open the connection between the filter 9 and the sealing cover 7, remove the filter 9, rotate the filter membrane limiter 902 to the slot 904 of the filter mesh 901, and remove the filter membrane to collect the cross-linked microcarriers deposited on the filter membrane.

[0050] The above description is merely a preferred embodiment of the present application and is not intended to limit the present application. Various modifications and variations are possible for those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present application shall be included within the scope of protection of the present application.

Claims

1. A microcarrier cross-linking cleaning system, characterized in that: The invention comprises a bracket (1) and a fixing frame (2) located above the bracket (1); a motor (3) is arranged inside the bracket (1); a tank body (4) is fixedly arranged on the fixing frame (2); a stirring device (5) is arranged inside the tank body (4); an output end of the motor (3) is connected to the stirring device (5); a liquid outlet pipe (6) is provided at the bottom of the tank body (4); a sealing cover (7) is provided at the top of the tank body (4); a feed port (8) and a filter (9) are connected on opposite sides of the sealing cover (7); the filter (9) is located inside the tank body (4) and above the stirring device (5).

2. The microcarrier cross-linking cleaning system according to claim 1, characterized in that: The filter (9) is a funnel-shaped filter.

3. The microcarrier crosslinking cleaning system according to claim 1, characterized in that: The filter (9) comprises a filter screen (901) and a filter membrane stopper (902) located inside, and a filter membrane is placed in the filter membrane stopper (902); a stopper plate (903) and a clamping slot (904) are provided on the top of the filter screen (901); the stopper plate (903) is detachably arranged on the sealing cover (7), and the filter membrane stopper (902) is rotatably clamped in the clamping slot (904).

4. The microcarrier cross-linking cleaning system according to claim 1, characterized in that: The stirring device (5) comprises a stirring shaft (501) and a stirring blade (502) connected to each other, and the stirring shaft (501) is connected to the output end of the motor (3).

5. The microcarrier cross-linking cleaning system according to claim 1, characterized in that: The sealing cover (7) is provided with at least one pair of the feed port (8) and the filter (9).

6. The microcarrier cross-linking cleaning system according to claim 1, characterized in that: A liquid inlet (11) is also provided on the top of the sealing cover (7), and the liquid inlet (11) is connected to the interior of the tank body (4).

7. The microcarrier cross-linking cleaning system according to claim 1, characterized in that: The liquid outlet pipe (6) is provided with a liquid outlet valve (10).

8. The microcarrier cross-linking cleaning system according to claim 1, characterized in that: A support column (12) is provided between the bracket (1) and the fixing frame (2).

9. The microcarrier crosslinking cleaning system according to any one of claims 1 to 8, characterized in that: The system further comprises a shell (13), wherein the shell (13) is located outside the bracket (1) and the tank body (4), and the sealing cover (7) and the feed port (8) are exposed at the top of the shell (13).

10. The microcarrier cross-linking cleaning system according to claim 9, characterized in that: The liquid outlet valve (10) on the liquid outlet pipe (6) is exposed outside the housing (13).

Images