CN210085426U

CN210085426U - Centrifugal sorting bioreactor

Info

Publication number: CN210085426U
Application number: CN201920581436.2U
Authority: CN
Inventors: 朱学仁; 朱羚
Original assignee: NANJING BO RUI BIOTECHNOLOGY Co Ltd
Current assignee: NANJING BO RUI BIOTECHNOLOGY Co Ltd
Priority date: 2019-04-26
Filing date: 2019-04-26
Publication date: 2020-02-18
Anticipated expiration: 2029-04-26

Abstract

The utility model discloses a centrifugal separation bioreactor, which comprises a feeding system, a centrifugal separation system and an imaging separation system; the feeding system comprises a peristaltic pump, a bubble sensor, a plurality of liquid storage bags and a pipe clamp valve; the centrifugal separation system comprises a centrifugal motor and a rotatable reaction tank; the centrifugal motor drives the reaction tank to rotate at a high speed around the central axis of the reaction tank to generate centrifugal force, and the substances in the reaction tank are centrifugally layered; the imaging sorting system comprises a micro camera, a stepping motor, a separating tube, a peristaltic pump, a bubble sensor, a liquid storage bag and a pipe clamp valve; the imaging sorting system can acquire the liquid layering condition in the reaction tank in real time; through control system contrast imaging, the stepper motor drives the separation tube to rotate, and the control system controls the peristaltic pump and the pinch valve in the pipeline to be opened and closed, so as to collect the required components in the layering. The utility model discloses directly realize centrifugal layering and partial shipment in bioreactor, easy operation avoids polluting.

Description

Centrifugal sorting bioreactor

Technical Field

The utility model relates to a centrifugal separation bioreactor belongs to bio-pharmaceuticals technical field, relates to the preparation of gomphosis antigen receptor T cell.

Background

The bioreactor is a container for culturing cells or tissues, the prior bioreactor does not realize centrifugal separation by autorotation, needs to be transferred into another centrifugal separation container, has troublesome operation, has pollution in the transfer process, occupies equipment, increases the cost of cleaning and disinfection and invests the equipment.

Disclosure of Invention

In order to solve the existing problems, the utility model discloses a centrifugal separation bioreactor. The utility model discloses provide a microscopic imaging sorting method simultaneously.

The utility model discloses a concrete technical scheme as follows:

the centrifugal separation bioreactor comprises a feeding system, a centrifugal separation system and an imaging separation system;

the feeding system comprises a peristaltic pump, a bubble sensor, a plurality of liquid storage bags and a pipe clamp valve;

the centrifugal separation system comprises a centrifugal motor and a rotatable reaction tank; the centrifugal motor drives the reaction tank to rotate at a high speed around the central axis of the reaction tank to generate centrifugal force, and the substances in the reaction tank are centrifugally layered;

the imaging sorting system comprises a micro camera, a stepping motor, a separating tube, a peristaltic pump, a bubble sensor, a liquid storage bag and a pipe clamp valve;

the imaging sorting system can acquire the liquid layering condition in the reaction tank in real time; the control system controls the opening and closing of the peristaltic pump and the pinch valve in the pipeline through contrast imaging.

The miniature camera is located above the shoulder, and collects layering information in the reaction tank vertically and downwards.

In the imaging separation system, the separating tube is arranged deviated from the center of the reaction tank, the upper end of the separating tube extends out of the reaction tank and is in transmission connection with the stepping motor, the lower end of the separating tube is L-shaped and bent, the stepping motor drives the separating tube to rotate around the central shaft, and the end face of the tube orifice of the separating tube and the wall of the reactor bottle are in concentric radian.

The bottom of the reaction tank is horizontal, a thickened base is arranged on the outer side of the bottom of the reaction tank, crossed grooves are formed in the thickened base, and the intersection point of the crossed grooves is located at the center of the thickened base;

a rotary tray is arranged below the thickening base in a contact manner, crossed cross convex strips are arranged on the upper surface of the rotary tray, the intersection points of the cross convex strips are located at the center of the rotary tray, and the cross convex strips are just matched and clamped in the cross grooves;

the centrifugal motor is arranged in the support, a central driving shaft of the motor is connected with the center of the rotating tray, a bearing is arranged between the rotating tray and the support, the motor drives the rotating tray to rotate, and the rotating tray and the reaction tank rotate synchronously.

The bearing comprises a bearing inner ring, a bearing outer ring and a plurality of balls arranged between the bearing inner ring and the bearing outer ring, the bearing outer ring is fixedly connected with the support and is not in contact with the rotating tray, and the bearing inner ring is fixedly connected with the rotating tray and is not in contact with the support.

The tank cover is connected with the tank opening through a rotary joint, and the tank cover is kept still when the reaction tank rotates.

A fixed upper cover is also arranged above the tank cover, a plurality of stabilizers are uniformly arranged on the circumference of the fixed upper cover,

the fixed upper cover is horizontally connected with a fixed upper supporting plate, the edge of the upper supporting plate is provided with a side plate which vertically extends downwards, and the lower end of the side plate is fixed with the bracket;

the fixed upper cover and the tank cover are evenly provided with a plurality of connecting holes in the same circumference, the connecting holes of the fixed upper cover and the connecting holes of the tank cover are coaxial in sequence, and a manual bolt is arranged in each connecting hole in a penetrating mode.

Microscopic imaging centrifugal sorting method, which is based on the centrifugal sorting bioreactor and is carried out in the centrifugal sorting bioreactor, comprises the following operation steps:

step 1: feeding: starting a peristaltic pump and a pipe clamp valve corresponding to the feeding pipeline, adding the plasma in the liquid storage bag into the tank, and stopping feeding when a bubble sensor in the feeding pipeline detects bubbles; after the preferable blood plasma is added into the tank, the PBS solution is injected into the blood plasma storage bag, the blood plasma bag is cleaned, the feeding step is repeated, and the cleaned PBS solution is continuously added into the tank; then a peristaltic pump and a pipe clamp valve in a corresponding pipeline of the sorting liquid storage bag are opened, and sorting liquid in a certain proportion is added into the tank;

step 2: group separation and centrifugation: starting a centrifugal motor, driving the reaction tank to rotate around the longitudinal central axis of the reaction tank by the centrifugal motor, and forming concentric circular ring shapes of all components in the reaction tank after the rotating speed of the centrifugal motor reaches a set value, wherein the thicknesses of all layers are consistent in the axial direction of the reaction tank;

step 3: and (3) confirming that layering is completed: collecting layered images vertically downwards on the shoulder through a miniature camera, comparing the layered images with preset layered images in a microscopic imaging centrifugal sorting system, and determining that boundaries of layers are clear, namely, determining that layering is finished;

and 4, step 4: starting the sub-suction mechanism: the stepping motor is started to drive the separation tube to do circular motion, and when the pipe orifice of the L section of the separation tube contacts the innermost layer, the peristaltic pump is started, and the separation tube starts to suck; a peristaltic pump and a pipe clamp valve in a pipeline corresponding to the waste liquid collecting bag are opened, and pumped liquid is pumped into the waste liquid collecting bag;

and 5: collecting useful component layers: when the separation tube rotates to quickly pump a useful component layer, the system automatically switches the corresponding peristaltic pump and the switch of the tube clamp valve through imaging contrast, and the useful component is collected into the liquid storage bag; when the useful components are collected and other layers are useless component layers, stopping the motor, stopping the reaction tank from rotating, and allowing all the components in the reaction tank to fall to the bottom of the tank;

step 6: discharge of useless components: the liquid falling into the reaction tank will be pumped into the waste bag by the peristaltic pump through the drain pipe (the reaction tank is preferably pumped into the PBS solution wash tank again and discharged through the drain pipe).

And 7: the control system controls the peristaltic pump in the pipeline for reversely rotating the useful components, opens the pipe clamp valve in the corresponding pipeline and conveys the useful components back to the tank.

The layers of the useful components are preset by a computer, and according to the preset layers, the system can automatically open and switch the peristaltic pump and the pipe clamp valve in the response pipeline.

The utility model discloses a theory of operation is:

the utility model discloses, start the motor of bioreactor below, motor drive bioreactor jar body is high-speed rotatory, suspension in the bioreactor is by centrifugal layering, form a plurality of layers that the circumference is adjacent in proper order in the retort, gather layering information through miniature camera machine, whether it is clear to judge whether centrifugal complete according to the layering looks connection face, after complete centrifugation, the layering is accomplished, keep the rotational speed unchangeable, start step motor rotating separation pipe, different layering compositions of suction from inside to outside in proper order, after centrifugal separation, the useful composition that will collect is carried back to in the clean jar once more.

The utility model has the advantages that:

the utility model realizes centrifugation in the bioreactor, does not need to replace the container, has shorter time and operates totally-enclosed full-automatic operation.

Drawings

FIG. 1 is a schematic structural diagram of the present invention (also referred to as abstract figure);

FIG. 2 is a schematic cross-sectional view of FIG. 1, illustrating the sorting process of the present invention;

reference numerals: 1-reaction tank, 2-layering, 3-tank cover, 4-fixed upper cover, 5-connecting hole, 6-subpackaging bag, 7-pipe clamp valve, 8-micro camera, 9-separating pipe, 10-cross groove, 11-cross raised line, 12-support, 13-motor, 14-bearing, 15-thickening base, 16-rotary tray and 17-stabilizer.

Detailed Description

The invention will be further elucidated with reference to the drawings and the detailed description. It should be understood that the following detailed description is illustrative of the invention only and is not intended to limit the scope of the invention.

The first embodiment is as follows:

the centrifugal separation bioreactor mainly comprises two parts, one is a centrifugal separation system, and the other is an imaging separation system; the culture product in the bioreactor is first centrifuged to separate the components 2 by centrifugation and then the separately separated components are absorbed into different separate bags 6. When layering 2 for the different composition in the centrifugation process, the thickness is unanimous around retort 1's circumference for the layering 2 of same layer, and retort 1 is vertical to be put, and the jar mouth is upwards, and the tank bottoms is downwards, and the tank bottoms is provided with the actuating mechanism that can drive retort 1 around its central axis is rotatory. The overall structure is shown in fig. 1.

Example two:

further, a mechanism for realizing centrifugal rotation is designed as follows: a motor 13 arranged below the reaction tank 1, wherein the motor 13 is positioned in the bracket 12, a central driving shaft of the motor 13 is connected with the center of the rotating tray 16, a bearing 14 is arranged between the rotating tray 16 and the bracket 12, the bearing 14 comprises a bearing 14 inner ring and a bearing 14 outer ring, and a plurality of balls arranged between the bearing 14 inner ring and the bearing 14 outer ring, the bearing 14 outer ring is fixedly connected with the bracket 12 and is not contacted with the rotating tray 16, and the bearing 14 inner ring is fixedly connected with the rotating tray 16 and is not contacted with the bracket 12; the motor 13 rotates the rotary tray 16, and the rotary tray 16 rotates in synchronization with the reaction tank 1.

Example three:

a connecting mechanism of the reaction tank 1 and the rotary tray 16 is further designed, in order to improve the connecting stability of the reaction tank 1 and the rotary tray 16, the bottom of the reaction tank 1 is horizontal, a thickened base 15 is arranged on the outer side of the bottom of the reaction tank 1, a cross-shaped groove 10 is formed in the thickened base 15, and the intersection point of the cross-shaped groove 10 is located at the center of the thickened base 15; the contact of the below of thickening base 15 is provided with rotatory tray 16, and the upper surface of rotatory tray 16 is equipped with criss-cross protruding strip 11, and the nodical of criss-cross protruding strip 11 is located the center of rotatory tray 16, and criss-cross protruding strip 11 just coincide the card in cross recess 10. The cross convex strips 11 and the cross concave grooves 10 are mutually embedded. The reaction tank 1 and the rotary tray 16 can be stably connected in the rotating process.

Example four:

the stable coupling mechanism of cover 3 has further been designed, in order to strengthen the stability of cover 3, the top of cover 3 still is provided with fixed upper cover 4, fixed upper cover 4 all around the outer cover 3 all around, fixed upper cover 4 circumference evenly is provided with a plurality of stabilizer 17. The stabilizer 17 serves to stabilize the pressure of fixing the upper cover 4.

A plurality of connecting holes 5 are uniformly formed in the circumferences of the fixed upper cover 4 and the tank cover 3, the connecting holes 5 of the fixed upper cover 4 and the connecting holes 5 of the tank cover 3 are sequentially coaxial, a pneumatic shaft penetrates through each connecting hole 5, and the periphery of the pneumatic shaft is sealed with the connecting holes 5. The pneumatic shaft plays a role in fixing and fixedly connecting the fixed upper cover 4 and the tank cover 3.

It is also possible to compact an upper support plate on the upper surface of the fixed upper lid 4, which is connected to the side of the bracket 12 through a side plate, to secure the can lid 3.

A plurality of connecting holes 5 are uniformly formed in the equal circumferences of the fixed upper cover 4 and the tank cover 3, the connecting holes 5 of the fixed upper cover 4 and the connecting holes 5 of the tank cover 3 are sequentially coaxial, and a manual bolt is arranged in each connecting hole 5 in a penetrating mode.

EXAMPLE five

The shape of the tank opening is further designed, the tank opening is reduced to form a horizontal shoulder, the microscopic imaging centrifugal separation system comprises a micro-camera 8 positioned above the shoulder, and the micro-camera 8 vertically and downwards acquires information of the layering 2 in the reaction tank 1. Whether the centrifugation is complete or not is judged through the acquisition information of the miniature camera 8, when the layering 2 component is sucked out, the next layer is sucked, and the next packaging bag 6 is switched.

Example six:

the sub-suction mechanism is further designed and comprises a plurality of sub-packaging bags 6 and separation tubes arranged on a tank cover 3 of the reaction tank 1, the separation tubes 9 are in sterile connection with the sub-packaging bags 6 through multi-channel hoses, tube clamp valves 7 and peristaltic pumps are arranged on the multi-channel hose pipelines, and the work of the tube clamp valves 7 and the peristaltic pumps is automatically controlled by a computer through an imaging system. Referring to fig. 2, the liquid inlet is opposite to each layer of liquid to be sucked, the separation tube 9 is installed on the tank cover 3, the separation tube is driven by the stepping motor 13 to rotate and move towards the inner wall of the reaction tank 1, and the separation tube 9 is installed at the position of the tank cover 3 deviated from the center of a circle.

Example seven:

a microscopic imaging centrifugal sorting system is further designed, and the microscopic imaging centrifugal sorting system can instantly collect the conditions of liquid layers 2 (the liquid layers 2 are sequentially supernatant, white blood cells and red blood cells from inside to outside) in the reaction tank 1; the microscopic imaging centrifugal sorting system is in cascade control with the pipe clamp valve and the peristaltic pump, and can respectively control the switch of each pipe clamp valve and the peristaltic pump to switch different subpackaging bags 6.

Example eight:

the utility model discloses the method includes: step 1: feeding: starting a peristaltic pump and a pipe clamp valve corresponding to the feeding pipeline, adding the plasma in the liquid storage bag into the tank, and stopping feeding when a bubble sensor in the feeding pipeline detects bubbles; after the preferable blood plasma is added into the tank, the PBS solution is injected into the blood plasma storage bag, the blood plasma bag is cleaned, the feeding step is repeated, and the cleaned PBS solution is continuously added into the tank; then a peristaltic pump and a pipe clamp valve in a corresponding pipeline of the sorting liquid storage bag are opened, and sorting liquid in a certain proportion is added into the tank;

and step 3: and (3) confirming that layering is completed: collecting layered images vertically downwards on the shoulder through a miniature camera, comparing the layered images with preset layered images in a microscopic imaging centrifugal sorting system, and determining that boundaries of layers are clear, namely, determining that layering is finished;

and step 3: starting the sub-suction mechanism: the stepping motor is started to drive the separation tube to do circular motion, and when the pipe orifice of the L section of the separation tube contacts the innermost layer, the peristaltic pump is started, and the separation tube starts to suck; a peristaltic pump and a pipe clamp valve in a pipeline corresponding to the waste liquid collecting bag are opened, and pumped liquid is pumped into the waste liquid collecting bag;

step 6: discharge of useless components: the liquid falling into the reaction tank is pumped into a waste liquid bag through a liquid outlet pipe by a peristaltic pump, and the reaction tank can be pumped into a PBS solution cleaning tank again and is discharged through the liquid outlet pipe;

The technical means disclosed by the scheme of the utility model is not limited to the technical means disclosed by the technical means, but also comprises the technical scheme formed by the arbitrary combination of the technical characteristics.

In light of the foregoing description of the preferred embodiments of the present invention, it is to be understood that various changes and modifications may be made by those skilled in the art without departing from the spirit and scope of the invention. The technical scope of the present invention is not limited to the content of the specification, and must be determined according to the scope of the claims.

Claims

1. The centrifugal separation bioreactor is characterized by comprising a feeding system, a centrifugal separation system and an imaging separation system;

the imaging sorting system can acquire the liquid layering condition in the reaction tank in real time; the control system is used for contrast imaging and controlling the opening and closing of the peristaltic pump and the pipe clamp valve in the pipeline and the rotation of the separation pipe.

2. The centrifugal separation bioreactor of claim 1, wherein the space between the top of the reaction tank and the opening of the tank is reduced radially to form a horizontal shoulder, and a micro-camera is positioned above the shoulder and vertically downward collects layering information in the reaction tank.

3. The centrifugal separation bioreactor as claimed in claim 2, wherein in the imaging separation system, the separation tube is disposed off-center of the reaction tank, the upper end of the separation tube extends out of the reaction tank and is in transmission connection with the stepping motor, the lower end of the separation tube is in an L-shaped bend, the stepping motor drives the separation tube to rotate around the central shaft, and the end surface of the tube orifice of the separation tube and the bottle wall of the reactor are in concentric radian.

4. The centrifugal separation bioreactor according to any one of claims 1 to 3, wherein the bottom of the reaction tank is horizontal, a thickened base is arranged on the outer side of the bottom of the reaction tank, the thickened base is provided with crossed cross grooves, and the intersection point of the cross grooves is positioned in the center of the thickened base;

the below contact of thickening base is provided with rotatory tray, the upper surface of rotatory tray is equipped with the criss-cross sand grip, and the nodical center that is located rotatory tray of cross sand grip, the card that just coincide of cross sand grip is in the cross recess.

5. The centrifugal separation bioreactor as claimed in claim 4, wherein a support is provided under the reaction tank, the centrifugal motor is located in the support, a central driving shaft of the motor is connected with the center of the rotating tray, a bearing is provided between the rotating tray and the support, the motor drives the rotating tray to rotate, and the rotating tray and the reaction tank rotate synchronously.

6. The centrifugal separation bioreactor of claim 5, wherein the bearing comprises an inner bearing ring and an outer bearing ring, and a plurality of balls disposed between the inner bearing ring and the outer bearing ring, the outer bearing ring is fixedly connected to the support frame and is not in contact with the rotating tray, and the inner bearing ring is fixedly connected to the rotating tray and is not in contact with the support frame; the reaction tank is provided with a tank cover, the tank cover is connected with the tank opening through a rotary joint, and the tank cover is kept still when the reaction tank rotates.

7. The centrifugal separation bioreactor as claimed in claim 6, wherein a fixed upper cover is further arranged above the tank cover, and a plurality of stabilizers are uniformly arranged on the circumference of the fixed upper cover;