In-situ drainage connector and in-situ drainage device
By designing the combination of in-situ drainage connector and filter components, the problems of low liquid discharge efficiency and high cost of filtration equipment of microcarrier bioreactors are solved, and efficient and low-cost liquid discharge and microcarrier protection are achieved.
Patent Information
- Application Number
- CN202520123240.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-20
- Publication Date
- 2025-02-25
- Estimated Expiration
- 2035-01-20
AI Technical Summary
Existing microcarrier bioreactors are inefficient during liquid discharge and replacement, and are prone to erroneous discharge of microcarriers. The filtration device is expensive and non-single-use, which poses a risk of contamination.
An in-situ liquid discharge connector is designed, adopting hollow structure and a piecewise structure jack, combined with an interference-matched filter component to achieve low-cost and efficient liquid discharge, avoiding the mis-discharge of microcarriers.
It improves the efficiency of liquid discharge and replacement, reduces dead volume, reduces costs, avoids the risk of damage and contamination of microcarriers, and is suitable for the production of actual biological samples.
Smart Images
Figure CN222524522U_ABST
Abstract
Description
Technical Field
[0001] The utility model relates to the technical field of microcarrier culture and bioreactor production, in particular to an in-situ liquid discharge connector and an in-situ liquid discharge device. Background Art
[0002] Microcarrier-based bioreactors (tanks) are the standard method for culturing adherent cells and producing biological products on a large scale. When the liquid in the tank (such as culture fluid or buffer, etc.) is to be discharged or replaced, the usual method is to wait for the microcarriers to settle naturally by gravity (the waiting time increases with the size of the tank), and then extract the supernatant through a drainage tube. In order to avoid sucking out the microcarriers and cells, there is a certain distance between the drainage tube mouth and the surface of the microcarrier sediment, which forms a dead volume for drainage. It is necessary to add the required replacement liquid (such as culture fluid or buffer, etc.) again, and wait for the microcarriers to settle again before draining. It is usually necessary to repeat the replacement process several times to completely replace the original liquid. In addition, a lower flow rate is required when draining to prevent the liquid flow from disturbing the microcarrier sediment and causing the microcarrier to be discharged by mistake. Therefore, the efficiency of liquid extraction and / or drainage (liquid replacement) of current microcarrier reactors is low.
[0003] Lowering the drain pipe (reducing dead volume) or increasing the drain flow rate can improve the drain efficiency, but in order to avoid the discharge of microcarriers, a filter device needs to be installed at the drain port to intercept the microcarriers. The current filter devices are mainly customized, expensive, and not disposable (there is a risk of contamination). Therefore, most bioreactors do not use filter interception devices.
[0004] In addition, for some special production process requirements (such as the production of engineered migration bodies), multiple replacement methods cannot be used, and only ectopic drainage can be used, that is, all microcarrier suspensions are discharged into the filter outside the tank to intercept the microcarriers and remove the culture medium in the tank (such as direct dumping), and the microcarriers in the filter are rinsed with replacement fluid (such as PBS), and then the filter is inverted and the microcarriers are rinsed back into the reactor with the replacement fluid. This drainage (replacement) efficiency is very high, but it causes a great impact on the cells, causing cell detachment, damage, and / or death, and is not suitable for the production of actual biological samples. Utility Model Content
[0005] In order to solve the above technical problems, the utility model provides an in-situ liquid discharge connector and an in-situ liquid discharge device including the in-situ liquid discharge connector, through which the in-situ liquid discharge device can discharge liquid in-situ at low cost and high efficiency. Specifically:
[0006] In a first aspect of the utility model, an in-situ drainage connector is provided, which is a hollow structure; a limiting protrusion is arranged in the hollow structure of the in-situ drainage connector; a plug hole is arranged at the upper end of the in-situ drainage connector, and the plug hole is formed by two or more slice structures surrounding it; one or more slots (for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) are arranged on the in-situ drainage connector, and an interference fit platform is arranged on the slot.
[0007] Preferably, the slot is arranged at the lower end and / or the side wall of the in-situ drainage connector; the slot is communicated with the hollow structure of the in-situ drainage connector.
[0008] It should be understood that the slot is connected to the hollow structure of the in-situ drainage connector, which means that holes are provided on the side wall of the in-situ drainage connector and / or the lower end of the in-situ drainage connector and the slot.
[0009] Preferably, the slots may be all arranged at the lower end of the in-situ drainage connector, or all arranged at the side wall of the in-situ drainage connector, or partly arranged at the lower end of the in-situ drainage connector and the other part arranged at the side wall of the in-situ drainage connector.
[0010] Preferably, the sliced structure is formed by extending outwardly from the upper end of the hollow structure of the in-situ drainage connector.
[0011] Preferably, the material of the sheet-type structure is an elastic material.
[0012] Preferably, the slice structure is relatively thin.
[0013] It should be understood that elastic materials, and / or thinner slice structures, are beneficial to the deformation of the slice structure.
[0014] Preferably, the shape of the segmented structure is a rectangle, a trapezoid or any other geometric shape suitable for providing friction, so as to facilitate the insertion and fixation of the insert (such as the drainage tube).
[0015] Preferably, the outer edge of the slice structure is arc-shaped.
[0016] Preferably, each sliced structure is independently arranged.
[0017] Preferably, the number of the slice structures is 6-20, more preferably 8-12, for example 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.
[0018] Preferably, the slice structure has at least one protrusion.
[0019] Preferably, the slice structure forms at least one raised portion inside the insertion hole, and the upper end of the raised portion is provided with a chamfer.
[0020] The second aspect of the utility model provides an in-situ drainage device, which includes the in-situ drainage connector described in the first aspect, and also includes a drainage pipe and / or a filter component; the drainage pipe is detachably installed in the hollow structure of the in-situ drainage connector through the socket of the in-situ drainage connector; the filter component is interference fit with the slot of the in-situ drainage connector.
[0021] In a specific embodiment of the present invention, the filter component and the slot of the in-situ drainage connector are interference-fitted via an interference-fit platform.
[0022] Furthermore, the slot is an arc-shaped slot matching the filter component.
[0023] It should be understood that the interference fit will not loosen by itself, and the interference fit connection method can withstand large axial and radial loads, provide a reliable fixing effect, reduce the risk of loosening due to vibration or impact, enhance the stability of the connection, provide a good sealing effect, and effectively prevent microcarriers from entering the in-situ drainage device.
[0024] Preferably, the number of the filtering components may be one or more, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more.
[0025] In a specific embodiment of the present utility model, the filter component is installed on the lower end and / or side wall of the in-situ drainage connector.
[0026] In a specific implementation of the present invention, the filter component is only installed at the lower end of the in-situ drainage connector.
[0027] In a specific embodiment of the present invention, the filter component is only installed on the side wall of the in-situ drainage connector. It should be noted that when the filter component is only installed on the side wall, the lower end of the hollow structure of the in-situ drainage connector needs to be closed to prevent microcarriers from entering the in-situ drainage device. Preferably, in order to achieve the technical effect of minimizing the dead volume, the filter component needs to be installed at the lower position of the side wall of the in-situ drainage connector.
[0028] Preferably, the liquid discharge tube has a scale for quantitative liquid extraction.
[0029] Preferably, the diameter of the hollow structure or the diameter of the plug hole of the in-situ drainage connector is greater than or equal to the diameter of the drainage pipe.
[0030] Preferably, the diameter of the hollow structure or the diameter of the plug hole of the in-situ drainage connector is 5-40 mm, preferably 10-20 mm, such as 5, 10, 15, 20, 25, 30, 35 or 40 mm.
[0031] Preferably, the drainage tube is fixed in the in-situ drainage connector via a limiting protrusion.
[0032] Preferably, the position of the drainage pipe relative to the in-situ drainage connector is determined by a limiting protrusion.
[0033] Preferably, the lower end of the drainage pipe is provided with a drainage port, the drainage port extends into the filter component provided at the lower end of the in-situ drainage connector, and is spaced from the filter screen at the bottom of the filter component. Preferably, the spacing is 1-20 mm, preferably 5-10 mm, for example 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 mm.
[0034] It should be understood that the structure of the drainage pipe can be a tubular structure of any specification, as long as it can be inserted into the hollow structure of the in-situ drainage connector and can be fixed at a specific position in the in-situ drainage connector (for example, a position with a certain distance from the filter screen at the bottom of the filter component) by a limiting protrusion.
[0035] In a specific embodiment of the present invention, the diameter of the lower end of the drainage pipe gradually decreases, and the diameter of the lowest end of the drainage pipe is the smallest, which is the drainage port.
[0036] It should be understood that in order to ensure that the microcarriers in the culture medium are prevented from entering the in-situ drainage device through the pores between the slice structures during drainage, the in-situ drainage device needs to have a certain height so that the plug hole (or slice structure) is located above the liquid surface of the culture medium. Usually, the height of the in-situ drainage device formed by assembling the in-situ drainage connector and the filter component is 3-50 cm, preferably 5-35 cm, such as 3, 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 cm.
[0037] Preferably, the in-situ drainage device is integrally formed, or is assembled from an independent in-situ drainage connector, a drainage pipe and a filter component.
[0038] The terms “include” or “comprising” described in the present invention are open-ended expressions, which include the specified components or steps described, as well as other specified components or steps that will not be substantially affected.
[0039] The term "and / or" in the present invention includes all combinations of items connected by the term, and each combination should be deemed to have been listed separately herein. For example, "A and / or B" includes "A", "A and B" and "B". For another example, "A, B and / or C" includes "A", "B", "C", "A and B", "A and C", "B and C" and "A and B and C".
[0040] Beneficial effects of the utility model:
[0041] 1. Provided is an in-situ drainage connector, comprising a socket formed by two or more slice structures, such a design can provide friction to facilitate the insertion and fixation of an insert (such as a drainage tube), and different socket opening diameters can be designed to adapt to drainage tubes of different sizes (including commercially available drainage tubes). The in-situ drainage connector is made of the same material from top to bottom and is integrally formed. When used to connect other components (such as a drainage tube and / or a filter component), the sealing of the device can be enhanced only by mechanical deformation and friction without the need for other connections (such as threaded connections, etc.).
[0042] 2. An in-situ drainage device including the in-situ drainage connector is provided, which can discharge liquid in situ while ensuring that the microcarriers remain in the culture tank; the liquid in the culture tank can be discharged more thoroughly, avoiding the problem of large dead volume during drainage in the prior art, and simplifying the complex process.
[0043] 3. A slot is provided at the lower end of the in-situ drainage connector, and an interference fit platform is provided on the slot. The filter component is interference fit with the slot and will not loosen by itself. The interference fit connection method can withstand large axial and radial loads, provide a reliable fixing effect, reduce the risk of loosening due to vibration or impact, enhance the stability of the connection, provide a good sealing effect, and effectively prevent microcarriers from entering the in-situ drainage connector or the in-situ drainage device.
[0044] 4. In the preferred embodiment, the drainage tube has a scale to enable quantitative sampling;
[0045] 5. In the preferred embodiment, the utility model can also be designed with slots of different sizes to adapt to filter components of different sizes (including commercially available filter components), thereby reducing costs and avoiding additional costs and pollution risks caused by maintenance such as cleaning. BRIEF DESCRIPTION OF THE DRAWINGS
[0046] Figure 1 : Schematic diagram of the structure of the in-situ drainage connector;
[0047] Figure 2 : Schematic diagram of the structure of the in-situ drainage device;
[0048] Figure 3 : Top view of the in-situ drainage device;
[0049] Figure 4 :In-situ drainage device along Figure 3 Sectional view at AA in the middle;
[0050] Figure 5 : Schematic diagram of the structure of an in-situ drainage device equipped with multiple filtering devices;
[0051] Figure 6 : A top view of an in-situ drainage device equipped with multiple filtering devices;
[0052] Figure 7 :In-situ drainage device with multiple filter devices Figure 6 Cross-section view at the middle BB.
[0053] The reference numerals in the figure are as follows: 1-in-situ drainage connector, 2-jack, 3-slot, 4-limiting protrusion, 5-piece structure, 6-chamfer, 7-drainage pipe, 8-filter component, 9-drainage port, 10-interference fit platform. DETAILED DESCRIPTION
[0054] The following will be combined with the drawings in the embodiments of the utility model to clearly and completely describe the technical solutions in the embodiments of the utility model. Obviously, the described embodiments are only some embodiments of the utility model, not all. Based on the embodiments in the utility model, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the utility model.
[0055] Example 1 An in-situ drainage connector
[0056] according to Figure 1 As shown, the in-situ drainage connector 1 is a hollow structure, and a limited position protrusion 4 is arranged in the hollow structure. The upper end of the in-situ drainage connector 1 is provided with a plug hole 2, and the plug hole 2 is formed by a slice structure 5 (the slice structure 5 is formed by the upper end of the hollow structure of the in-situ drainage connector 1 extending outward, and the slice structure 5 is conducive to the deformation of the insert (such as the drainage tube 7) when inserted). The slice structure 5 forms at least one protrusion inside the plug hole 2, and the upper end of the protrusion is provided with a chamfer 6. Each slice structure 5 is independently arranged, and the in-situ drainage connector 1 includes 6-20 slice structures 5 (preferably 8-12, and 10 in this embodiment). The lower end and / or side wall of the in-situ drainage connector 1 are provided with one or more slots 3 (in this embodiment, the slot is arranged at the lower end as an example), and the slot 3 is connected to the hollow structure of the in-situ drainage connector 1. The slot 3 is provided with an interference fit platform 10.
[0057] It should be understood that when the in-situ drainage connector 1 of the present invention is installed with the drainage pipe 7, the diameter of the hollow structure of the in-situ drainage connector 1 and / or the opening diameter of the jack 2 is greater than or equal to the diameter of the drainage pipe 7. The slice structure 5 is made of a thinner elastic material, which is conducive to deformation and facilitates the insertion of the drainage pipe 7.
[0058] Example 2 An in-situ drainage device
[0059] according to Figure 2 , Figure 3 and Figure 4 As shown, the drain pipe 7, the filter component 8 and the in-situ drain connector 1 are installed together to prepare the in-situ drain device. The filter component 8 is interference fit with the slot 3 below the in-situ drain connector 1, so that the filter component 8 is sealed and connected to the in-situ drain connector 1 through the slot 3 to prevent the microcarrier from entering the in-situ drain device during the drain process. The drain pipe 7 is inserted into the hollow structure of the in-situ drain connector 1 through the jack 2 to form a stable connection, and the insertion depth of the drain pipe 7 is limited by the limiting protrusion 4 until the drain port 9 below the drain pipe 7 is spaced a certain distance (for example, 1-20 mm, preferably 5-10 mm) from the filter screen at the bottom of the filter component 8. The assembly order is to assemble the filter component 8 first, and then assemble the drain pipe 7.
[0060] The diameter of the lower end of the drainage pipe 7 gradually decreases, and the diameter of the lowest end of the drainage pipe is the smallest, which is the drainage port 9.
[0061] like Figure 4 As shown, a limiting protrusion 4 is provided in the hollow structure of the in-situ drainage connector 1, and the limiting protrusion 4 limits the insertion depth of the drainage pipe 7, and adjusts the distance between the drainage port 9 below the drainage pipe 7 and the filter screen at the bottom of the filter component 8. The interference fit platform 10 provided on the slot 3 is used to achieve interference fit with the filter component 8, which plays a role of fastening and sealing.
[0062] The in-situ drainage device of the utility model has a certain height to ensure that the plug hole 2 (or the slice structure 5) is located above the liquid surface of the culture solution during drainage, so as to prevent the culture solution or the microcarriers in the culture solution from entering the in-situ drainage device through the plug hole 2 (or the gap between the slice structures 5). Generally, the height of the in-situ drainage device formed by assembling the in-situ drainage connector 1 and the filter component 8 is 3-50 cm (preferably 5-35 cm).
[0063] The in-situ liquid discharge device designed in this utility model has the following functions / characteristics:
[0064] 1. Discharge liquid in situ from a bioreactor (e.g., a culture tank);
[0065] 2. Quick installation;
[0066] 3. Single use;
[0067] 4. By using the in-situ drainage connector of the utility model, the distance between the drainage port below the drainage pipe and the bottom filter screen of the filter component can be adjusted to avoid clogging of the filter screen;
[0068] 5. Design different opening diameters of the jack to adapt to drainage pipes of different sizes;
[0069] 6. The discharge tube has scales to allow quantitative sampling. For example, the discharge tube is a pipette of different models (such as 10 mL, 25 mL, 50 mL pipettes).
[0070] Example 3 Another in-situ drainage device (equipped with multiple filtering devices)
[0071] The in-situ drainage device of the present invention can be equipped with multiple filter components 8 to increase the filtration area and / or drainage flux, and further increase the drainage efficiency while preventing the microcarriers from being discharged. It should be noted that the position, direction, and number of the filter components 8 are not limited by the accompanying drawings. In theory, as long as the in-situ drainage connector 1 is large enough, dozens or more filter components 8 can be installed at its lower end and / or side wall.
[0072] Figure 5 , Figure 6 and Figure 7 A schematic diagram showing an example of assembling three filter components 8 is shown.
[0073] It should be understood that in order to minimize the "dead volume", all filter components 8 can be arranged at the bottom of the in-situ drainage connector 1, and the openings of the slots 3 arranged on the in-situ drainage connector 1 are all outward to facilitate the assembly of the filter components.
[0074] Example 4 Effect test of the in-situ drainage device of the present application
[0075] Taking the in-situ liquid drainage device described in Example 2 as an example, the effect of the in-situ liquid drainage device described in the utility model is verified.
[0076] Specific methods: 1) In-situ drainage group: first use the in-situ drainage device of the utility model, add PBS and stir slightly, drain the liquid in-situ and then take samples; 2) Ectopic drainage group: pour out the entire culture system into the MCAC 70μm filter, rinse with PBS, and then rinse the microcarrier back into the tank with PBS (without affecting the function of the microcarrier) and then take samples. The two groups of samples were observed for cell shedding under the bright field of the microscope, and dead cells were observed by propidium staining.
[0077] The results showed that, whether after one drainage or multiple treatments (i.e., a series of multi-step drainage-addition eMig production processes), the cell shedding and number of dead cells on the microcarriers in the in situ drainage group were significantly less than those in the ectopic drainage group. For example, taking the 500 mL HEK293FT-Plastic microcarrier culture system as an example, after one drainage, more than 50% of the cells in the ectopic drainage group fell off after drainage, while only less than 10% of the cells in the in situ drainage group fell off. After multiple treatments, nearly 100% of the cells in the ectopic drainage group fell off, while only about 50% of the cells in the in situ drainage group fell off.
[0078] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of simple modifications, and these simple modifications all belong to the protection scope of the present invention.
[0079] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present utility model will not further describe various possible combinations.
Claims
1. An in-situ drainage connector, characterized in that: The in-situ drainage connector is a hollow structure; A limited position protrusion is arranged in the hollow structure of the in-situ drainage connector; The upper end of the in-situ drainage connector is provided with a plug hole, and the plug hole is formed by two or more slice structures surrounding each other; The in-situ drainage connector is provided with one or more slots, and the slots are provided with interference fit platforms.
2. The in-situ drainage connector according to claim 1, characterized in that: The slot is arranged at the lower end and / or the side wall of the in-situ drainage connector; the slot is communicated with the hollow structure of the in-situ drainage connector.
3. The in-situ drainage connector according to claim 1, characterized in that: The sliced structure is formed by extending outwardly from the upper end of the hollow structure of the in-situ drainage connector.
4. The in-situ drainage connector according to claim 1, characterized in that: The slice structure forms at least one protrusion inside the insertion hole, and the upper end of the protrusion is provided with a chamfer.
5. The in-situ drainage connector according to claim 1, characterized in that: The number of the slice structures is 6 to 20, and each slice structure is independently arranged.
6. An in-situ drainage device, characterized in that: The in-situ drainage device comprises the in-situ drainage connector according to any one of claims 1 to 5, and further comprises a drainage pipe and / or a filter component; The drainage tube is detachably installed in the hollow structure of the in-situ drainage connector through the socket of the in-situ drainage connector; The filter component is interference-fitted with the slot of the in-situ drainage connector.
7. The in-situ drainage device according to claim 6, characterized in that: The liquid discharge tube has a scale for quantitatively taking liquid.
8. The in-situ drainage device according to claim 6, characterized in that: The diameter of the hollow structure or the diameter of the plug hole of the in-situ drainage connector is greater than or equal to the diameter of the drainage pipe.
9. The in-situ drainage device according to claim 6, characterized in that: The drainage pipe is fixed in the in-situ drainage connector via a limiting protrusion.
10. The in-situ drainage device according to claim 9, characterized in that: The lower end of the drainage pipe is provided with a drainage port, and the drainage port extends into the filter component provided at the lower end of the in-situ drainage connector, and is spaced apart from the filter screen at the bottom of the filter component.