CN116218672A - Interception component, preparation method thereof and biological reaction bag - Google Patents

Abstract

The application provides a interception component, a preparation method thereof and a biological reaction bag, wherein the interception component comprises: the retaining piece comprises a retaining area and a frame area, wherein the retaining area is of a porous structure, and the frame area is of a solid structure; one end of the upper layer film is fixedly connected with the frame area of the interception piece; the connecting bag buckle is fixedly connected with one end of the upper membrane far away from the interception piece and forms a containing cavity together with the interception piece and the upper membrane; wherein the material of the interception piece comprises a high molecular polymer, and the molecular weight of the high molecular polymer is more than or equal to 10 ⁶ . The upper membrane is connected after the frame area of the interception piece is solidified, so that the problem of leakage of intercepted microorganisms can be effectively prevented; and adopts high molecular polymerThe manufacturing of the interception piece can improve the rigidity and bending resistance of the interception piece, and avoid the problem of breakage and cracking in the use process.

Description

Interception component, preparation method thereof and biological reaction bag

Technical Field

The application relates to the field of biology, in particular to a interception component, a preparation method thereof and a biological reaction bag.

Background

Culturing microorganisms such as cells in disposable bioreactor bags may involve perfusion, also known as "continuous cell culture", which involves a fluid transfer process, i.e., providing a steady source of fresh cell culture medium to the bioreactor vessel, and constantly removing waste products and/or spent culture medium from the bioreactor vessel. In the dynamic perfusion process, two points need to be ensured, namely, the liquid culture medium can smoothly circulate, and microorganisms such as cells and the like need to be kept in the environment of the liquid culture medium as much as possible. Therefore, the disposable bioreactor bag for continuous cell culture needs to have a member for achieving the retention of substances.

Currently, the trapping parts on the market mostly adopt porous polymer sheets, and the penetration strength of the pore structure is about 72N/mm around the polymer sheets. In the actual swing reaction microorganism culture process, the interception part has lower mechanical property, and is easy to crack when uncontrollable bending and collision occur, so that the intercepted microorganisms and other substances leak, and further economic property loss is caused. The pore structure around the interception part causes the problem that the joint of the interception part and the upper layer film is not easy to be closed when the interception part is combined with the upper layer film, and the joint of the upper layer film and the interception part is easy to cause leakage of intercepted microorganisms in the actual microorganism culture process.

It should be noted that the information disclosed in the foregoing background section is only for enhancement of understanding of the background of the present application and thus may include information that does not form the prior art that is already known to those of ordinary skill in the art.

Content of the application

Aiming at the problems in the prior art, the application aims to provide a interception component and a preparation method thereof, which can effectively prevent the problems of leakage of intercepted microorganisms and easy breakage and cracking of interception parts.

An embodiment of the present application provides a trapping assembly, comprising: the retaining piece comprises a retaining area and a frame area, wherein the frame area is arranged around the retaining area, the retaining area is of a porous structure, and the frame area is of a solid structure; the upper layer of the membrane is provided with a plurality of layers,one end of the upper membrane is fixedly connected with the frame area of the interception piece; the connecting bag buckle is fixedly connected with one end of the upper membrane far away from the interception piece and forms a containing cavity together with the interception piece and the upper membrane; wherein the material of the interception piece comprises a high molecular polymer, and the molecular weight of the high molecular polymer is more than or equal to 10 ⁶ 。

In some embodiments, the high molecular weight polymer comprises ultra-high molecular weight polyethylene.

In some embodiments, the penetration strength of the retention element is greater than or equal to 300N/mm.

In some embodiments, the pore size of the porous structure of the entrapment zone is less than the diameter of the microorganism being entrapped.

In some embodiments, the cross-sectional shape of the retentate perpendicular to its thickness comprises at least one of a circle, square, triangle, trapezoid, or irregular pattern.

In some embodiments, the retention zone of the retention member is recessed away from the upper membrane to form at least one retention chamber.

In some embodiments, the surface of the retaining member adjacent to the side of the connecting pocket includes at least one of a flat surface, a curved surface, or a cambered surface.

In some embodiments, the bezel area has a width ranging between 1mm and 500 mm.

In some embodiments, the securing of the upper membrane to the border region of the retentate comprises hot melt welding.

In some embodiments, the upper film is coated on the outer surface of the frame area.

In some embodiments, a spacer is disposed within the receiving cavity.

In some embodiments, the material of the spacer comprises a sponge, a water absorbent resin, or other hydrophilic material.

The embodiment of the application also provides a preparation method of the interception component, which comprises the following steps: providing a retaining piece, wherein the retaining piece comprises a retaining area and a frame area; carrying out solidification treatment on the frame area of the interception piece; thermally welding an upper membrane to the border region of the retentate; and a connecting bag buckle is fixed at one end of the upper layer film far away from the interception piece.

In some embodiments, in the step of providing a retention element, comprising: and pouring the ultra-high molecular weight polyethylene powder into a metal mold, and increasing the temperature and the pressure of the metal mold to sinter the ultra-high molecular weight polyethylene powder into the interception piece.

In some embodiments, the ultra high molecular weight polyethylene powder has a particle size ranging between 10 μm and 100 μm.

In some embodiments, the solidifing process comprises: and filling the frame area by using sealant, sealing and welding the frame area by using polymer melt, and/or melting the frame area by using a high-temperature pressurizing method.

The embodiment of the application also provides a bioreactor bag, comprising: a reaction bag body including a waste outlet; and the interception component can be arranged in the reaction bag body to intercept microorganisms in the reaction bag body; wherein the connecting bag buckle is communicated with the waste outlet through a connecting pipe.

In some embodiments, the reaction pouch body further comprises: the sampling port is used for acquiring trapped microorganisms in the reaction bag body at different time intervals; a liquid infusion port for adding the culture liquid in the reaction bag body; an exhaust port for exhausting at least part of the gas in the reaction bag body; a pressure regulating port for regulating the pressure in the reaction bag body; and an output port for obtaining the trapped microorganisms after the completion of the cultivation.

The interception component provided by the application has the following advantages:

the interception area is of a porous structure, so that the exchange of culture solutions inside and outside the interception component can be realized, and the frame area is of a solid structure, so that on one hand, the sealing performance of the frame area of the interception piece can be improved; on the other hand, the stable connection of the upper membrane and the interception piece is facilitated, and the problem that the intercepted microorganisms are easy to leak at the joint of the interception piece and the upper membrane is effectively prevented.

Further, the material of the interception piece comprises a high molecular polymer, and the molecular weight of the high molecular polymer is more than or equal to 10 ⁶ The rigidity and bending resistance of the interception piece can be greatly improved, and the problem that the interception piece is damaged and cracked in the using process is avoided.

Drawings

Other features, objects and advantages of the present application will become more apparent upon reading of the detailed description of non-limiting embodiments, made with reference to the following drawings.

FIG. 1 is a schematic cross-sectional view of a retention assembly according to an embodiment of the present application;

FIG. 2 is a schematic perspective view of a retaining assembly according to an embodiment of the present application;

FIG. 3 is a side view of the entrapment assembly of FIG. 2;

FIG. 4 is a schematic perspective view of a second embodiment of the retaining assembly;

FIG. 5 is a side view of the entrapment assembly of FIG. 4;

FIG. 6 is a schematic perspective view of a trapping assembly according to an embodiment of the present application;

FIG. 7 is a side view of the entrapment assembly of FIG. 6;

FIG. 8 is a schematic perspective view of a retaining assembly according to an embodiment of the present application;

FIG. 9 is a side view of the entrapment assembly of FIG. 8;

FIG. 10 is a schematic perspective view of a retaining assembly according to an embodiment of the present application;

FIG. 11 is a side view of the hold-down assembly of FIG. 10;

FIG. 12 is a schematic cross-sectional view of another alternative embodiment of the retention assembly described in the examples herein;

FIG. 13 is a schematic view of a partial structure of a retention assembly according to an embodiment of the present application;

FIG. 14 is a schematic view of a portion of a second embodiment of the entrapment component;

FIG. 15 is a schematic cross-sectional view of another alternative embodiment of the retention assembly described in the examples herein;

FIG. 16 is a flow chart of the preparation of the retention assembly described in the examples of the present application

FIG. 17 is a schematic view of the structure of the entrapment component in the preparation step S1 in the embodiment of the present application;

FIG. 18 is a schematic view of the structure of the entrapment component in the preparation step S2 in the embodiment of the present application;

FIG. 19 is a schematic view of the structure of the entrapment component in the preparation step S3 in the embodiment of the present application;

FIG. 20 is a schematic view of the structure of the entrapment component in the preparation step S4 in the embodiment of the present application;

FIG. 21 is a schematic cross-sectional view of a bioreactor bag according to an embodiment of the present application.

Reference numerals:

1. biological reaction bag

10. Interception component

110. Retaining piece

111. Interception area

112. Frame area

113. Interception cavity

120. Upper layer film

130. Connecting bag buckle

140. Accommodating cavity

150. Spacer member

20. Reaction bag body

21. Waste outlet

22. Sampling port

23. Liquid infusion port

24. Exhaust port

25. Pressure regulating port

26. Output port

Detailed Description

Example embodiments will now be described more fully with reference to the accompanying drawings. However, the exemplary embodiments can be embodied in many forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the concept of the example embodiments to those skilled in the art. The same reference numerals in the drawings denote the same or similar structures, and thus a repetitive description thereof will be omitted. "or", "or" in the specification may each mean "and" or ". Although the terms "upper", "lower", "between", etc. may be used in this specification to describe various example features and elements of the present application, these terms are used herein for convenience only, e.g., in terms of the directions of the examples in the drawings. Nothing in this specification should be construed as requiring a particular three-dimensional orientation of the structure to fall within the scope of this application. Although the terms "first" or "second" etc. may be used herein to describe certain features, these features should be interpreted in a descriptive sense only and not for purposes of limitation as to the number and importance of the particular features.

The structure of the entrapment assembly 10 in various embodiments is described in detail below with reference to the figures. It is to be understood that the drawings and the following descriptions are exemplary only and are not intended to limit the scope of the present application.

Referring to fig. 1, an embodiment of the present application provides a retention assembly 10 comprising: the retaining piece 110, the retaining piece 110 comprises a retaining area 111 and a frame area 112, the frame area 112 is arranged around the retaining area 111, the retaining area 111 is of a porous structure, and the frame area 112 is of a solid structure; an upper membrane 120, wherein one end of the upper membrane 120 is fixedly connected with the frame region 112 of the interception member 110; the connecting bag buckle 130 is fixedly connected with one end of the upper membrane 120 far away from the interception piece 110, and forms a containing cavity 140 together with the interception piece 110 and the upper membrane 120; wherein the material of the retention element 110 comprises a high molecular polymer having a molecular weight of greater than or equal to 10 ⁶ . The connection bag buckle 130 is communicated with the outside, so that the replacement of the culture solution of the trapped microorganisms is realized. The entrapment zone 111 is porous to entrap the entrapped microorganisms and prevent them from passing through the attached bagThe button 130 leaks to the outside; the frame area 112 is of a solid structure, so that the leakage problem of trapped microorganisms can be effectively prevented; and the high molecular polymer is adopted to manufacture the interception piece 110, so that the rigidity and bending resistance of the interception piece 110 can be improved, and the problem of breakage and cracking in the use process is avoided. Preferably, the periphery of the upper membrane 120 is fixedly connected with the retaining member 110, the middle part of the upper membrane is provided with an opening, and the connecting bag buckle 130 is arranged at the opening and is fixed with the upper membrane 120.

The interception area 111 is a porous structure, and preferably, the pore diameter of the porous structure of the interception area 111 is smaller than the diameter of the intercepted microorganisms, so that the intercepted microorganisms can be intercepted, and the intercepted microorganisms are prevented from leaking to the outside through the connecting bag buckle 130. It will be appreciated that the pore size of the retention zone 111 is measured in accordance with ASTM E1294-89, preferably by a BSD-PBL full-automatic pore size analysis tester using a bubble pressure method using an immersion liquid of 60v.% IPA/H ₂ O, the test temperature is normal temperature; the diameter of the trapped microorganisms is measured by a conventional method, and is not described in detail in the present application. The frame region 112 is a solid structure, and includes filling the frame region 112 with a sealant, sealing and welding the frame region 112 with a polymer melt, and/or melting the frame region 112 with a high-temperature pressurizing method.

Preferably, the sealing glue is used for filling the border region 112, so that the border region 112 forms a solid structure, and on one hand, the sealing performance of the border region 112 of the retaining piece 110 can be improved; on the other hand, the sealant can pave the surface of the interception piece 110 in the frame area 112, so that the flatness of the surface of the interception piece 110 connected with the upper membrane 120 is improved, the stable connection of the upper membrane 120 and the interception piece 110 is facilitated, and the problem that the intercepted microorganisms are easy to leak at the joint of the interception piece 110 and the upper membrane 120 is effectively prevented.

Preferably, the material of the sealant comprises a resin comprising at least one of a polyester, a polyolefin, a polysulfone, or a polyamide. The resin material is selected, and the fluidity is utilized, so that the void structure on at least the surface of the retaining member 110 can be well filled, and after the resin material is solidified, the outer surface of the frame area 112 of the retaining member 110 forms a flat surface, thereby facilitating the stable combination of the upper membrane 120 and the retaining member 110.

The material of the retention element 110 comprises a high molecular polymer having a molecular weight of greater than or equal to 10 ⁶ The rigidity and bending resistance of the retaining piece 110 can be greatly improved, and the problem that the retaining piece 110 is damaged and cracked in the use process is avoided. It is understood that high molecular weight polymers refer to high molecular weight compounds formed by repeated linkages, including crystalline, amorphous, oriented, and woven structures. Preferably, the high molecular polymer has a molecular weight in the range of 10 ⁶ To 10 ⁸ Between, for example: the molecular weight of the high molecular polymer was 10 ⁷ But is not limited thereto. It can be appreciated that in the process of biological cultivation, the material and structure of the retaining member 110 in the present application can ensure that the retained microorganisms are always located outside the retaining component 10, and the waste of the cultivation of the retained microorganisms is discharged through the connection bag opening 130 of the retaining component 10, so as to achieve the purpose of retaining the retained microorganisms.

In some embodiments, the high molecular weight polymer comprises ultra high molecular weight polyethylene. The ultra-high molecular weight polyethylene has ultra-strong wear resistance and rigidity, good bending resistance, stable chemical property and strong ageing resistance, and can well avoid physical damage caused by mechanical external forces such as collision, bending and the like in the culture process compared with the material of the retaining piece 110 used at present.

In some embodiments, the puncture strength of the retaining member 110 is greater than or equal to 300N/mm, so that the puncture resistance of the retaining member 110 is ensured, the puncture resistance is the relative ability of the material to prevent tearing from progressing after being pierced by the incision or the score, and during use of the retaining assembly 10, due to actions such as collision and shaking, the retained microorganisms may exert a certain puncture force on the retaining member 110, so that stress is relatively concentrated at a specific position, and due to the greater puncture strength of the retaining member 110 compared with the current material, the problem of stress concentration in the specific region caused by the actions such as collision and shaking can be effectively resisted, so that the service life of the retaining member 110 is prolonged. Preferably, the penetration strength of the retention element 110 ranges between 300N/mm and 800N/mm, for example: 400N/mm, 450N/mm, 500N/mm, 550N/mm, 600N/mm, 650N/mm, 700N/mm, 750N/mm.

Referring to fig. 2-7 in combination, in some embodiments, the cross-sectional shape of the retention element 110 perpendicular to its thickness includes at least one of a circle, square, triangle, trapezoid, or irregular pattern. The cross-sectional shape of the retaining member 110 in the direction perpendicular to the thickness direction of the retaining member 110 in fig. 2 to 3 is circular, the cross-sectional shape of the retaining member 110 in the direction perpendicular to the thickness direction of the retaining member 110 in fig. 4 to 5 is square, the cross-sectional shape of the retaining member 110 in the direction perpendicular to the thickness direction of the retaining member 110 in fig. 6 to 7 is hexagonal, and the cross-sectional shape of the retaining member 110 in the direction perpendicular to the thickness direction of the retaining member is not limited by fig. 2 to 7, but may be other shapes, and the description of the cross-sectional shape is omitted. It will be appreciated that it is only necessary to ensure that the junction of the retentate 110 and the upper membrane 120 is filled with sealant.

Referring to fig. 8 to 11 in combination, in some embodiments, the interception area 111 of the interception member 110 is recessed away from the upper membrane 120 to form at least one interception chamber 113, and the interception chamber 113 is configured to increase the surface area of the interception area 111, so as to prevent the problem of slow liquid discharge rate caused by too small surface area of the interception area 111 being covered by the intercepted microorganisms; and due to the existence of the interception cavity 113, the biological reaction bag can be supported to a certain extent, so that the problem that the intercepted microorganisms break by themselves due to extrusion caused by volume compression when the liquid in the biological reaction bag is replaced is prevented. For example: in fig. 8 to 9, the retaining member 110 includes a retaining chamber 113, and the specific shape of the space of the retaining chamber 113 is not limited to fig. 8 to 9; it will be appreciated that the provision of the entrapment cavity 113 prevents the upper membrane 120 from adhering to the inner surface of the entrapment piece 110 when entrapping the entrapped microorganisms, resulting in a problem of reduced rate when replacing the liquid in the bioreactor bag.

In fig. 10 to 11, the interception member 110 includes four interception chambers 113, and when there are a plurality of interception chambers 113, the specific spatial shape of each interception chamber 113 is not limited to fig. 8 to 9, and the spatial shapes of the interception chambers 113 on the same interception member 110 may be set differently. By the arrangement, the supporting function of the biological reaction bag can be further improved, and the problem that the biological reaction bag is broken by itself due to extrusion caused by the fact that the volume of the biological reaction bag is compressed when the liquid in the biological reaction bag is replaced is solved. And also prevents the upper membrane 120 from being attached to the inner surface of the entrapping element 110, resulting in a problem of a decrease in the rate of replacement of the liquid in the bioreactor bag. Further, the different interception areas 111 of the interception member 110 are arranged in different porous structures and correspond to different interception chambers 113; that is, the pores of the walls of each of the interception chambers 113 are different in size, so that the components of substances allowed to pass through are different, and the waste generated during cultivation of the intercepted microorganisms can be distributed and discharged from different interception chambers according to the particle size of the waste, and at this time, the interception path of one interception chamber 113 can be opened or closed by the upper membrane 120, so that the different waste generated can be classified and recovered. The interception path of the upper membrane 120 to open or close the interception chamber 113 can be achieved by a different connection relationship with the interception member 110, for example: if the interception path of a certain interception chamber 113 is to be closed, the interception path can be realized by fixedly connecting the interception chamber with the upper membrane 120 near the wall surface of one side of the upper membrane 120.

Referring to fig. 12 in combination, in some embodiments, the surface of the retaining member 110 adjacent to the side of the connecting pocket button 130 includes at least one of a flat surface, a curved surface, or a curved surface. It can be understood that the surface of the retaining member 110 near the side of the connecting bag buckle 130 may be configured according to different usage situations, and is not limited to the surface structure in fig. 12, which is not repeated herein.

In some embodiments, the width of the bezel area 112 ranges between 1mm and 500 mm. Preferably, the width of the rim region 112 ranges between 5mm and 10mm, for example: 6mm, 7mm, 8mm, 9mm, etc., but is not limited thereto. This arrangement ensures effective bonding between the upper membrane 120 and the retentate 110, and prevents cracking at the junction of the upper membrane 120 and the retentate 110 due to the too narrow width of the rim region 112.

Referring to fig. 13, in some embodiments, the attachment of the upper membrane 120 to the rim region 112 of the retaining member 110 includes hot melt welding. That is, the upper membrane 120 and the rejection 110 are stably combined by heating to a designated temperature between the upper membrane 120 and the rejection 110 and then pressurizing. It should be understood that hot melt welding is only one way to connect the upper membrane 120 to the retaining member 110 as proposed in the present application, and is not intended to limit the connection of the present application, for example: the upper membrane 120 may be fixedly connected to the frame region 112 of the retaining member 110 by, but not limited to, adhesive connection.

Referring to fig. 14, in some embodiments, the upper film 120 is coated on the outer surface of the frame region 112. That is, the upper membrane 120 extends from the surface of the side edge region 112 of the retaining member 110 near the side of the connection pocket 130, through the side surface of the retaining member 110, until the side edge region 112 of the retaining member 110 is far away from the side surface of the connection pocket 130, until the whole of the side edge region 112 of the retaining member 110 is covered, so as to achieve the fixed connection between the upper membrane 120 and the retaining member 110. On one hand, the sealant is filled in the frame area 112 of the retaining piece 110, so that the sealing performance of the frame area 112 of the retaining piece 110 is improved; on the other hand, by coating the frame area 112, secondary sealing is realized, and the sealing performance of the frame area 112 is further improved; this double-seal arrangement can better prevent the problem of leakage of trapped microorganisms at the junction of the trap 110 and the upper membrane 120.

Referring to fig. 15 in combination, in some embodiments, a spacer 150 is disposed in the receiving chamber 140. The material of the spacer comprises sponge, water-absorbent resin or other hydrophilic materials. The spacer 150 can further improve the supporting effect on the bioreactor bag, and prevent the problem that the bioreactor bag is broken by itself due to extrusion caused by the trapped microorganisms due to volume compression when the liquid in the bioreactor bag is replaced. And also prevents the upper membrane 120 from being attached to the inner surface of the entrapping element 110, resulting in a problem of a decrease in the rate of replacement of the liquid in the bioreactor bag, thereby functioning as a buffer.

Referring to fig. 16 to 20 in combination, the embodiment of the present application further provides a method for preparing the interception component 10, which includes the following steps:

referring to fig. 16 to 17, step S1: a retention element 110 is provided, the retention element 110 comprising a retention region 111 and a border region 112, the border region 112 being disposed around the retention region 111.

In step S1, the method for preparing the retention element 110 comprises: the ultra-high molecular weight polyethylene powder is poured into a metal mold, and the temperature and pressure of the metal mold are increased to sinter the ultra-high molecular weight polyethylene powder into the entrapping part 110. The particle size of the ultra-high molecular weight polyethylene powder is between 10 μm and 100 μm; preferably, the ultra-high molecular weight polyethylene powder has a particle size in the range of 40 μm to 60 μm, for example: the particle size of the ultra-high molecular weight polyethylene powder was in the range of 50. Mu.m.

Referring to fig. 16 to 18, step S2: the border region 112 of the retention element 110 is solidifiable.

In step S2, the solidifing process includes: the frame area is filled with sealant, and/or sealed and welded with polymer melt, and/or melted by high-temperature pressurizing. Preferably, the rim region 112 is filled with a sealant such that the rim region 112 forms a solid structure. The material of the sealant comprises a resin, and the resin comprises at least one of polyester, polyolefin, polysulfone or polyamide. The resin material is selected, and the fluidity is utilized, so that the void structure on at least the surface of the retaining member 110 can be well filled, and after the resin material is solidified, the outer surface of the frame area 112 of the retaining member 110 forms a flat surface, thereby facilitating the stable combination of the upper membrane 120 and the retaining member 110.

Referring to fig. 16 to 19, step S3: thermally welding the upper membrane 120 to the border region 112 of the retention element 110; the hot melt welding is only one way to attach the upper membrane 120 to the retention element 110 as set forth herein and is not intended to limit the manner in which the present application is attached.

In step S3, it is preferable that the upper film 120 is heat-welded to the outer surface of the rim region 112 of the rejection member 110 while the rim region 112 is heat-welded to the upper film 120. That is, during the thermal welding process of the upper film 120, the upper film 120 extends along a side surface of the retaining member 110 near the connecting pocket 130 to a side surface of the retaining member 110 far from the connecting pocket 130, so as to cover the upper film 120 on the outer surface of the frame region 112. The sealing performance of the frame area 112 of the interception piece 110 is improved by filling the frame area 112 of the interception piece 110 with sealant; on the other hand, by coating the frame area 112, secondary sealing is realized, and the sealing performance of the frame area 112 is further improved; this double-seal arrangement can better prevent the problem of leakage of trapped microorganisms at the junction of the trap 110 and the upper membrane 120.

Referring to fig. 16 to 20, step S4: a connecting bag buckle 130 is fixed at one end of the upper membrane 120 far away from the interception piece 110; the connecting bag buckle 130 is fixedly connected with one end of the upper membrane 120 far away from the interception piece 110, and forms a containing cavity 140 together with the interception piece 110 and the upper membrane 120, and is communicated with the outside through the connecting bag buckle 130, so that liquid substance exchange in the interception component 10 is realized.

Referring to fig. 21 in combination, the present application further provides a bioreactor bag 1, comprising: a reaction pouch body 20 and a entrapment assembly 10, the reaction pouch body 20 including a waste outlet 21; the interception component 10 can be arranged in the reaction bag body 20 to intercept microorganisms in the reaction bag body 20; wherein the connection bag buckle 130 of the interception component 10 is communicated with the waste outlet 21 through a connection pipe so as to realize the discharge of waste and the replacement of culture solution in the biological reaction bag 1.

In some embodiments, the reaction pouch body 20 further comprises: a sampling port 22 for acquiring trapped microorganisms at different time intervals within the reaction pouch body 20; a liquid infusion port 23 for adding the culture liquid in the reaction bag body 20; a gas outlet 24 for discharging at least part of the gas in the reaction bag body 20; a pressure regulating port 25 for reducing the pressure in the reaction bag body 20; and an output port 26 for taking out the trapped microorganisms after completion of the cultivation. It can be understood that the sampling port 22, the liquid infusion port 23, the exhaust port 24, the pressure regulating port 25 and the output port 26 are all disposed on the reaction bag body 20 and connected to the outside through connection pipes to achieve respective functions.

It should be understood that the shape of the bioreactor bag 1 may be any shape, and the waste outlet 21, the sampling port 22, the liquid infusion port 23, the air outlet 24, the pressure regulating port 25, and the output port 26 may be disposed at any position on the reactor bag body 20 of the bioreactor bag 1, which is not limited in fig. 21.

Several preferred embodiments of the present application are described below:

example 1

The present embodiment provides a entrapment assembly 10 comprising: a retention element 110, an upper membrane 120 and a connecting bag button 130; the retaining member 110 comprises a retaining area 111 and a frame area 112, wherein the frame area 112 is arranged around the retaining area 111, and the frame area 112 is filled with sealant to seal the frame area 112; one end of the upper membrane 120 is fixedly connected with the surface of the side frame region 112 of the interception piece 110, which is close to one side of the connecting bag buckle 130, and the other end of the upper membrane is fixedly connected with the connecting bag buckle 130 and forms a containing cavity 140 together with the interception piece 110; the liquid substance exchange in the interception component 10 is realized through the connection bag buckle 130 communicated with the outside. Wherein the material of the interception piece 110 adopts ultra-high molecular weight polyethylene with the molecular weight of more than or equal to 10 ⁶ 。

Example 2

The present embodiment provides a entrapment assembly 10 comprising: a retention element 110, an upper membrane 120 and a connecting bag button 130; the retaining member 110 comprises a retaining area 111 and a frame area 112, wherein the frame area 112 is arranged around the retaining area 111, and the frame area 112 is filled with sealant to seal the frame area 112; one end of the upper membrane 120 extends from the side surface of the side frame region 112 of the retaining member 110, which is close to the side surface of the connecting bag buckle 130, through the side surface of the retaining member 110 until the side frame region 112 of the retaining member 110 is far away from the side surface of the connecting bag buckle 130, until the whole side frame region 112 of the retaining member 110 is covered, so as to realize the fixed connection between the upper membrane 120 and the retaining member 110, and the other end is fixedly connected with the connecting bag buckle 130 and encloses a containing cavity 140 together with the retaining member 110; the liquid substance exchange in the interception component 10 is realized through the connection bag buckle 130 communicated with the outside. Wherein the material of the retaining member 110 is selected fromWith ultra-high molecular weight polyethylene having a molecular weight of 10 or more ⁶ 。

Example 3

The present embodiment provides a entrapment assembly 10 comprising: a retention element 110, an upper membrane 120 and a connecting bag button 130; the retaining member 110 comprises a retaining area 111 and a frame area 112, the retaining area 111 of the retaining member 110 is recessed away from the upper membrane 120 to form at least one retaining cavity 113, and the retaining cavity 113 and the accommodating cavity 140 together provide a space environment required by biological culture; the frame region 112 is disposed around the interception region 111, and the frame region 112 is filled with sealant to seal the frame region 112; one end of the upper membrane 120 is fixedly connected with the surface of the side frame region 112 of the interception piece 110, which is close to one side of the connecting bag buckle 130, and the other end of the upper membrane is fixedly connected with the connecting bag buckle 130 and forms a containing cavity 140 together with the interception piece 110; the liquid substance exchange in the interception component 10 is realized through the connection bag buckle 130 communicated with the outside. Wherein the material of the interception piece 110 adopts ultra-high molecular weight polyethylene with the molecular weight of more than or equal to 10 ⁶ 。

Example 4

The present embodiment provides a bioreactor bag 1 comprising: the reaction bag body 20 and the interception component 10, wherein the reaction bag body 20 comprises a waste outlet 21, a sampling 22, a liquid infusion 23, an exhaust port 24, a pressure regulating port 25 and an output port 26; the waste outlet 21 is used for realizing the discharge of waste and the replacement of culture solution in the biological reaction bag 1, the sampling port 22 is used for acquiring trapped microorganisms in the reaction bag body 20 at different time intervals, and the liquid infusion port 23 is used for adding culture solution in the reaction bag body 20; the exhaust port 24 is used for exhausting at least part of gas in the reaction bag body 20, the pressure regulator 25 is used for regulating the pressure in the reaction bag body 20, and the output 26 is used for acquiring trapped microorganisms after the culture is completed; the entrapment component 10 can be disposed within the reaction pouch body 20 to entrap microorganisms within the reaction pouch body 20.

The entrapment component 10 comprises: a retention element 110, an upper membrane 120 and a connecting bag button 130; the retaining member 110 includes a retaining region 111 and a frame region 112, the frame region 112 being disposed around the retaining region 111, the frameThe region 112 is filled with a sealant to seal the border region 112; one end of the upper membrane 120 is fixedly connected with the surface of the side frame region 112 of the interception piece 110, which is close to one side of the connecting bag buckle 130, and the other end of the upper membrane is fixedly connected with the connecting bag buckle 130, and forms a containing cavity 140 together with the interception piece 110, and a spacer 150 is arranged in the containing cavity 140; the liquid substance exchange in the interception component 10 is realized through the connection bag buckle 130 communicated with the outside. Wherein the material of the interception piece 110 adopts ultra-high molecular weight polyethylene with the molecular weight of more than or equal to 10 ⁶ 。

The connection bag buckle 130 of the interception component 10 is communicated with the waste outlet 21 through a connection pipe so as to realize the discharge of waste and the replacement of the culture solution in the biological reaction bag 1.

In summary, embodiments of the present application provide a retention assembly 10 comprising: the retaining piece 110, the retaining piece 110 comprises a retaining area 111 and a frame area 112, the frame area 112 is arranged around the retaining area 111, the retaining area 111 is of a porous structure, and the frame area 112 is of a solid structure; an upper membrane 120, wherein one end of the upper membrane 120 is fixedly connected with the frame region 112 of the interception member 110; the connecting bag buckle 130 is fixedly connected with one end of the upper membrane 120 far away from the interception piece 110, and forms a containing cavity 140 together with the interception piece 110 and the upper membrane 120; wherein the material of the retention element 110 comprises a high molecular polymer having a molecular weight of greater than or equal to 10 ⁶ . The liquid substance exchange in the interception component 10 is realized through the connection bag buckle 130 communicated with the outside. The interception area 111 is of a porous structure, so that the exchange of the culture solution inside and outside the interception component 10 can be realized, and the frame area 112 is of a solid structure, so that on one hand, the sealing performance of the frame area 112 of the interception piece 110 can be improved; on the other hand, the stable connection of the upper membrane 120 and the interception member 110 is facilitated, and the problem that the intercepted microorganisms are easy to leak at the joint of the interception member 110 and the upper membrane 120 is effectively prevented. Further, the material of the interception member 110 includes a high molecular polymer having a molecular weight of 10 or more ⁶ The rigidity and bending resistance of the retaining piece 110 can be greatly improved, and the problem that the retaining piece 110 is damaged and cracked in the use process is avoided.

The foregoing is a further detailed description of the present application in connection with the specific preferred embodiments, and it is not intended that the practice of the present application be limited to such description. It should be understood that those skilled in the art to which the present application pertains may make several simple deductions or substitutions without departing from the spirit of the present application, and all such deductions or substitutions should be considered to be within the scope of the present application.

Claims (18)

1. A retention assembly, comprising:

the retaining piece comprises a retaining area and a frame area, wherein the frame area is arranged around the retaining area, the retaining area is of a porous structure, and the frame area is of a solid structure;

one end of the upper layer film is fixedly connected with the frame area of the interception piece;

the connecting bag buckle is fixedly connected with one end of the upper membrane far away from the interception piece and forms a containing cavity together with the interception piece and the upper membrane; wherein the method comprises the steps of

The material of the interception piece comprises high molecular polymer, and the molecular weight of the high molecular polymer is more than or equal to 10 ⁶ 。

2. The retention assembly of claim 1, wherein the high molecular weight polymer comprises an ultra-high molecular weight polyethylene.

3. The retention assembly of claim 1, wherein the retention element has a puncture strength greater than or equal to 300N/mm.

4. The entrapment assembly of claim 1 wherein the pore size of the porous structure of the entrapment zone is less than the diameter of the microorganism to be entrapped.

5. The retention assembly of claim 1, wherein the cross-sectional shape of the retention member perpendicular to its thickness comprises at least one of a circle, square, triangle, trapezoid, or irregular pattern.

6. The retention assembly of claim 1, wherein the retention zone of the retention member is recessed away from the upper membrane to form at least one retention chamber.

7. The retention assembly of claim 1, wherein a surface of the retention member proximate the side of the connecting pocket comprises at least one of a planar surface, a curved surface, or a cambered surface.

8. The entrapment component of claim 1 wherein the border region has a width ranging from 1mm to 500 mm.

9. The retention assembly of claim 1, wherein the secured attachment of the upper membrane to the rim region of the retention member comprises a hot melt weld.

10. The retention assembly of claim 1, wherein the upper membrane is wrapped around an outer surface of the border region.

11. The retention assembly of claim 1, wherein a spacer is disposed within the receiving cavity.

12. The retention assembly of claim 10, wherein the material of the spacer comprises a sponge, a water absorbent resin, or other hydrophilic material.

13. A method of making a retention assembly according to any one of claims 1 to 12, comprising the steps of:

providing a retaining piece, wherein the retaining piece comprises a retaining area and a frame area;

carrying out solidification treatment on the frame area of the interception piece;

thermally welding an upper membrane to the border region of the retentate;

and a connecting bag buckle is fixed at one end of the upper layer film far away from the interception piece.

14. The method of claim 13, wherein the step of providing a retaining member comprises:

and pouring the ultra-high molecular weight polyethylene powder into a metal mold, and increasing the temperature and the pressure of the metal mold to sinter the ultra-high molecular weight polyethylene powder into the interception piece.

15. The method of manufacturing a trapping assembly according to claim 14, wherein the ultra-high molecular weight polyethylene powder has a particle size ranging between 10 μm and 100 μm.

16. The method of making a trapping assembly according to claim 13, wherein the solidifying process comprises: and filling the frame area by using sealant, sealing and welding the frame area by using polymer melt, and/or melting the frame area by using a high-temperature pressurizing method.

17. A bioreactor bag, comprising:

a reaction bag body including a waste outlet; and

the entrapment assembly of any of claims 1-12 wherein the entrapment assembly is positionable within the reaction pouch body to entrap microorganisms within the reaction pouch body; wherein the method comprises the steps of

The connecting bag buckle is communicated with the waste outlet through a connecting pipe.

18. The bioreactor bag of claim 17, wherein the reactor bag body further comprises:

the sampling port is used for acquiring trapped microorganisms in the reaction bag body at different time intervals;

a liquid infusion port for adding the culture liquid in the reaction bag body;

an exhaust port for exhausting at least part of the gas in the reaction bag body;

a pressure regulating port for regulating the pressure in the reaction bag body; and

and an output port for obtaining the trapped microorganisms after the completion of the cultivation.

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