CN216964638U - Horizontal self-stirring in-vitro protein synthesis reactor - Google Patents
Horizontal self-stirring in-vitro protein synthesis reactor Download PDFInfo
- Publication number
- CN216964638U CN216964638U CN202220216939.1U CN202220216939U CN216964638U CN 216964638 U CN216964638 U CN 216964638U CN 202220216939 U CN202220216939 U CN 202220216939U CN 216964638 U CN216964638 U CN 216964638U
- Authority
- CN
- China
- Prior art keywords
- horizontal
- stirring
- tank body
- protein synthesis
- self
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000003756 stirring Methods 0.000 title claims abstract description 98
- 230000014616 translation Effects 0.000 title claims abstract description 84
- 238000001243 protein synthesis Methods 0.000 title claims abstract description 81
- 238000000338 in vitro Methods 0.000 title claims abstract description 76
- 239000012295 chemical reaction liquid Substances 0.000 claims abstract description 39
- 238000000034 method Methods 0.000 claims abstract description 9
- 230000008569 process Effects 0.000 claims abstract description 5
- 230000005540 biological transmission Effects 0.000 claims description 9
- 230000007246 mechanism Effects 0.000 claims description 9
- 238000003466 welding Methods 0.000 claims description 8
- 238000009826 distribution Methods 0.000 claims description 6
- 238000007789 sealing Methods 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 4
- 230000000149 penetrating effect Effects 0.000 claims description 3
- 238000006243 chemical reaction Methods 0.000 abstract description 38
- 230000000694 effects Effects 0.000 abstract description 24
- 230000003321 amplification Effects 0.000 abstract description 2
- 238000003199 nucleic acid amplification method Methods 0.000 abstract description 2
- 239000007789 gas Substances 0.000 description 21
- 108090000623 proteins and genes Proteins 0.000 description 11
- 102000004169 proteins and genes Human genes 0.000 description 10
- 238000002156 mixing Methods 0.000 description 8
- 238000004113 cell culture Methods 0.000 description 4
- 238000005096 rolling process Methods 0.000 description 4
- 230000007480 spreading Effects 0.000 description 4
- 238000003892 spreading Methods 0.000 description 4
- 108010048367 enhanced green fluorescent protein Proteins 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000002194 synthesizing effect Effects 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000013519 translation Methods 0.000 description 3
- 230000009471 action Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 230000005484 gravity Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 238000013518 transcription Methods 0.000 description 2
- 230000035897 transcription Effects 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000003137 locomotive effect Effects 0.000 description 1
- 238000001823 molecular biology technique Methods 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 238000005498 polishing Methods 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 239000002912 waste gas Substances 0.000 description 1
Images
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J4/00—Feed or outlet devices; Feed or outlet control devices
- B01J4/001—Feed or outlet devices as such, e.g. feeding tubes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J19/28—Moving reactors, e.g. rotary drums
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
Abstract
The utility model discloses a horizontal self-stirring in-vitro protein synthesis reactor, which comprises a rack, a horizontal tank body, a tank cover assembly and a driving device, wherein the tank cover assembly comprises a tank cover and a tank cover; the horizontal tank body is rotatably arranged on the frame, the tank cover assembly is detachably sealed on at least one end part of the horizontal tank body, and the driving device is used for driving the horizontal tank body to rotate; the inner wall of the horizontal tank body is provided with a self-stirring paddle component, and the self-stirring paddle component is suitable for stirring the protein synthesis reaction liquid in the horizontal tank body in the rotating process of the horizontal tank body. The utility model can fully mix the reaction solution, improve the reaction efficiency, keep the activity of the product and carry out industrial amplification.
Description
Technical Field
The utility model belongs to the technical field of biological reaction equipment, and particularly relates to a horizontal self-stirring in-vitro protein synthesis reactor.
Background
Current methods for protein synthesis can be divided into two categories: the first is the traditional cell culture method for synthesizing protein, and the second is the in vitro cell-free method for synthesizing protein. The conventional cell culture method refers to a molecular biology technique for expressing foreign genes by model organisms such as bacteria, fungi, plant cells or animal cells. The in vitro cell-free method for synthesizing the protein mainly depends on a cell-free expression system, exogenous target DNA is used as a protein synthesis template, and substances such as a substrate required by protein synthesis and auxiliary factors of transcription and translation related proteins are artificially controlled and supplemented, so that the synthesis of the target protein can be realized. The protein expressed in the in vitro translation system does not need to be subjected to the steps of plasmid construction, transformation, cell culture, cell collection and crushing, and is a relatively quick, time-saving and convenient protein expression mode.
However, since the preparation of In vitro protein synthesis system is complicated and the addition of accessory factors is complicated, all In vitro protein synthesis products In the market are low In activity and extremely expensive (the cost is 100-1000 times that of the traditional cell culture method), and only very individual laboratories can prepare proteins with high activity by using a small amount of IVTT (In vitro transcription and translation) reaction solution at present. The reactor adopted in the preparation of the laboratory is a 24-hole plate, and the added IVTT reaction liquid is uL grade.
By taking the uL-grade reaction of a 24-pore plate as a contrast, when the reactor is replaced by a container with larger volume, such as a culture dish with the diameter of 16cm, a plastic basin with the diameter of 20 x 26cm, a plastic basin with the diameter of 30 x 40cm, a 1L beaker, an L-grade micro tube, an L-grade vertical reaction tank and the like, and the mL-grade or L-grade IVTT reaction solution is added for carrying out amplification reaction, the activity of the finally obtained product is greatly reduced, and the industrial practical value is not high. Therefore, it is not feasible to perform size enlargement only from the perspective of the volume of the reactor, and it is highly desirable to develop an industrial enlarging apparatus which can enlarge the volume greatly and maintain the activity of the final product to the maximum.
SUMMERY OF THE UTILITY MODEL
The utility model aims to solve the technical problems that the in vitro protein synthesis can only be completed in a small amount in a laboratory at present and cannot be amplified on an industrial scale.
In order to achieve the above objects, in a first aspect, the present invention provides a horizontal self-stirring in vitro protein synthesis reactor, comprising a frame, a horizontal tank, a tank cover assembly and a driving device; the horizontal tank body is rotatably arranged on the machine frame, the tank cover assembly is detachably sealed on at least one end part of the horizontal tank body, and the driving device is used for driving the horizontal tank body to rotate; the inner wall of the horizontal tank body is provided with a self-stirring paddle component, and the self-stirring paddle component is suitable for stirring the protein synthesis reaction liquid in the horizontal tank body in the rotating process of the horizontal tank body.
The tank cover assembly is provided with an air inlet channel and an air outlet channel, wherein the air inlet channel is communicated with an air outlet of the air source device; the tank cover assembly comprises a moving part which is suitable for rotating along with the rotation of the horizontal tank body.
Further, the height of the inner wall of the tank body protruding from the stirring blade assembly accounts for no more than 60% of the radius of the inner circle of the horizontal tank body.
Further, the self-stirring paddle component comprises a plurality of paddles, each paddle extends along the axial direction of the horizontal tank body, and the paddles are distributed at intervals along the circumferential direction of the inner circle of the horizontal tank body.
Further, the blades are arranged at equal intervals.
Furthermore, each paddle is integrally formed and fixed on the inner wall of the horizontal tank body through welding, and the welding seam is polished.
Furthermore, one side of each blade, which is far away from the inner wall of the horizontal tank body, is of a concave arc structure.
Furthermore, an included angle a is formed between the length direction of each blade and the axial direction of the horizontal tank body.
Further, the included angle a is less than or equal to 20 degrees.
Furthermore, an included angle b is formed between the profile surface of each blade and a tangent plane of the horizontal tank body passing through the inner edge of the blade, and the included angle b is less than 90 degrees.
Furthermore, the blade is characterized in that a plurality of through holes are distributed on the contour surface of each blade.
Further, the total area of the through holes on each blade accounts for 30% -50% of the area of the blade, and the distance between every two adjacent through holes is equal.
Further, the edge of each blade and the edge of the orifice of each through hole have a rounded corner structure.
Further, the volume of the horizontal tank body is 10L-10000L.
Furthermore, a heating device with adjustable temperature is arranged on the air inlet channel so as to adjust the temperature of air entering the horizontal tank body.
Further, the air source device is an air compressor or a fan.
Further, the moving part is a rotary joint, the air inlet channel is communicated with the horizontal tank body through the air inlet rotary joint, and the air outlet channel is communicated with the horizontal tank body through the air outlet rotary joint.
Furthermore, the tank cover assembly is of an integrated structure and is arranged at the feed inlet of the horizontal tank body.
Further, the cover assembly comprises a cover body and a gas distribution device penetrating through the center of the cover body, the gas distribution device comprises a gas inlet pipe and a gas outlet pipe, the gas inlet pipe extends into the horizontal tank body, and the gas outlet pipe is communicated with the horizontal tank body through a gas outlet hole formed in the cover body.
Further, the exhaust hole has a plurality and surrounds the intake pipe.
Furthermore, one end of the air inlet pipe, which extends into the horizontal tank body, is closed, and a plurality of air outlet holes are formed in the pipe wall of the air inlet pipe.
Furthermore, the air outlet holes are uniformly distributed along the circumferential direction of the air inlet pipe and extend along the axial direction of the air inlet pipe.
Furthermore, the length of the air inlet pipe extending into the horizontal tank body is equivalent to that of the horizontal tank body.
Further, the tank cover assembly is of a split structure and comprises an air inlet portion and an air outlet portion, the air inlet portion is mounted at the left end of the horizontal tank body, and the air outlet portion is mounted at the right end of the horizontal tank body.
Further, the air inlet part comprises an air inlet pipe, and the air inlet pipe extends into the horizontal tank body; the exhaust part comprises a cover body and an exhaust pipe, and the exhaust pipe is communicated with the horizontal tank body through an exhaust hole formed in the cover body.
Further, a sealing ring, a tank cover lock catch and a locking handle are arranged on the tank cover assembly, a tank body lock catch is arranged on the horizontal tank body, and the locking handle is suitable for locking the tank cover lock catch on the tank body lock catch.
Furthermore, one end of the horizontal tank body is supported by a bearing seat arranged on the rack, and the other end of the horizontal tank body is supported by a pair of rotating wheels arranged on the rack.
Further, the driving device is a motor, and the motor drives the horizontal tank body through a belt wheel transmission mechanism or a gear transmission mechanism.
Further, the axial direction of the horizontal tank body is parallel to the horizontal plane.
Compared with the prior art, the utility model has the following beneficial effects:
1. the reactor main body adopts a horizontal rotatable tank structure, a customized self-stirring paddle component is arranged on the inner wall of a tank cavity, fresh air is continuously introduced into the tank cavity, the volume of reaction liquid added into the tank cavity only accounts for a small part of the volume of the tank body, so that in the reaction process, on one hand, the reaction liquid can be thinned on the wall of the tank cavity through the rotation of the tank body, the thinning degree can be adjusted by controlling the volume of the added reaction liquid, the thinned reaction liquid can be more fully contacted with the air, and the reaction efficiency is improved; on the other hand, the self-stirring paddle component on the inner wall of the rotating tank body can automatically stir the reaction liquid, and because of the design of the included angle of the paddle and the concave arc structure with wide two ends and narrow middle, multidirectional surge flow and vortex flow can be formed, so that the reaction liquid can be fully mixed, and the reaction efficiency is improved.
2. The in vitro protein synthesis test is carried out on a plurality of reactors with different tank body volumes, the tank body volume is minimum 10L and maximum 10000L, the activity of a final product of each reactor is measured, the activity is equivalent to that of laboratory preparation, and the in vitro protein synthesis reactor has industrial popularization value, is an efficient, convenient and low-cost in vitro protein synthesis reactor and synthesis method, is undoubtedly a steam locomotive for promoting future biological industry revolution, scientific and technological progress, medicine research and development and medicine production, and has revolutionary effect on the whole biological industry.
Drawings
FIG. 1 is a first schematic perspective view of a reactor according to one embodiment of the present invention;
FIG. 2 is a schematic perspective view of a second embodiment of the reactor of the present invention;
FIG. 3 is a left side view of one embodiment of a reactor of the present invention;
FIG. 4 is a cross-sectional view of one embodiment of a reactor of the present invention;
FIG. 5 is a perspective view of the interior of a tank cavity of one embodiment of the reactor of the present invention;
FIG. 6 is a schematic perspective view of a tank lid assembly in one embodiment of the reactor of the present invention;
FIG. 7 is a schematic structural view of another embodiment of the reactor of the present invention;
fig. 8 is a graph showing the results of an activity test of EGFP, a target protein produced using one embodiment of the present invention.
In the drawings, a frame 1; a horizontal tank body 2; a self-stirring blade assembly 20; a paddle 21; a tank body lock 22; a rolling rib 23; a can lid assembly 3; a lid body 30; an exhaust vent 301; an intake pipe 31; an intake rotary joint 32; an exhaust pipe 33; an exhaust rotary joint 34; a tank cover lock catch 35; a locking handle 36; a heating device 37; a drive device 4; a bearing seat 5; a rotating wheel 6; a pulley transmission mechanism 7; and a fan 8.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The utility model is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
As shown in fig. 1 to 6, one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the present invention comprises a frame 1, a horizontal tank body 2, a tank cover assembly 3 and a driving device 4; the horizontal tank body 2 is rotatably arranged on the frame 1, the tank cover assembly 3 is detachably sealed on at least one end part of the horizontal tank body 2, and the driving device 4 is used for driving the horizontal tank body 2 to rotate; the inner wall of the horizontal tank body 2 is provided with a self-stirring paddle component 20, and the self-stirring paddle component 20 is suitable for stirring the protein synthesis reaction liquid in the horizontal tank body 2 in the rotating process.
In the embodiment, the horizontal tank body 2 is adopted to replace a common vertical reaction tank, the common independent vertical stirring paddle in the vertical reaction tank is replaced by a continuous wall type horizontal stirring paddle which is fixed with the inner wall of the tank body into a whole, the reaction liquid can be thinned on the wall of the tank body through the rotation of the tank body, the thinning degree can be adjusted by controlling the volume of the added reaction liquid, the thinned reaction liquid has larger surface area, and can be more fully contacted with air, so that the reaction efficiency is improved. In the conventional vertical reaction tank, reaction liquid is deposited at the bottom of the tank under the action of gravity, and even though stirring is carried out continuously by a stirring paddle, the thickness of the reaction liquid is basically unchanged, namely, the stirring cannot achieve the effect of spreading the reaction liquid to enable the reaction liquid to be in more full contact with air. The structure of the present embodiment can be said to have overturned the consistent practice in the art.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor, the horizontal self-stirring in-vitro protein synthesis reactor further comprises an air source device, wherein the tank cover assembly 3 is provided with an air inlet channel and an air outlet channel, and the air inlet channel is communicated with an air outlet of the air source device; the cover assembly 3 includes a moving portion adapted to rotate with the rotation of the horizontal can body 2. The air source device can control the external air to be conveyed to the horizontal tank body 2, can adjust the pressure, time and the like of air supply, and can adjust according to different reaction liquids so as to provide more optimized conditions for reaction.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the present invention, as shown in fig. 4, the height of the self-stirring paddle assembly 20 protruding from the inner wall of the tank body accounts for no more than 60% of the radius of the inner circle of the horizontal tank body. Preferably, the ratio is 50% or less. The height of the self-stirring paddle component 20 is not too high, the specific height can be customized according to the volume of the reaction liquid in the tank, and the more the reaction liquid is, the larger the height is, so that the stirring can be performed more fully. Since the reaction solution is thinned, the volume of the reaction solution does not exceed a certain ratio of the volume of the tank, and accordingly, the height of the self-stirring blade assembly 20 is limited.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the utility model, as shown in fig. 5, the self-stirring paddle assembly 20 comprises a plurality of paddles 21, each paddle 21 extends along the axial direction of the horizontal tank 2, and each paddle 21 is arranged at intervals along the circumferential direction of the inner circle of the horizontal tank 2. 6 paddles 21 that set up at an interval are shown in fig. 5, can divide into 6 regions with the jar chamber like this, and during operation, along with the rotation of horizontal jar body 2, under the combined action of centrifugal force, gravity and blade stirring power, reaction liquid can be separately shakeout in each region, because every region is enclosed by two paddles 21 and jar wall and is synthesized, reaction liquid can be shakeout to three face in this region, and the reaction liquid of same volume can be shakeout thinner like this, and the area of contact with the air is also bigger, improves reaction efficiency. The quantity of paddle 21 can be increased and decreased according to the volume size of horizontal jar of body 2, and general volume is big more, and the quantity of paddle 21 is more to can divide into a plurality of regions that the volume is less, be favorable to the shakedown of reaction solution.
In one embodiment of the horizontal self-stirring in vitro protein synthesis reactor of the present invention, as shown in FIG. 5, the blades 21 are arranged at equal intervals. The 6 equally spaced blades 21 can divide the tank cavity into 6 areas with equal volume, so that the spreading degree of the reaction liquid in each area is equivalent, namely equivalent to the controlled contact area, the reaction efficiency is uniform, and the whole reaction effect is better.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the utility model, each blade 21 is integrally formed and fixed on the inner wall of the horizontal tank body 2 by welding, and the welding seam is polished. The paddle 21 is integrally formed, has a smooth surface, and is fixed on the tank wall by welding, and the polishing treatment of the welding line can improve the smooth finish of the surface, reduce the obstruction to the flow of the reaction liquid, accelerate the mixing of the reaction liquid, and improve the reaction efficiency.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the present invention, as shown in fig. 5, one side of each of the blades 21 away from the inner wall of the horizontal tank 2 is a concave arc structure. As is apparent from fig. 5, the paddle 21 has two wide ends and a narrow middle, and the two ends are smoothly transited through the concave arc, so that the reaction solution can conveniently flow between the divided regions, and the overall mixing efficiency is improved.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the present invention, the length direction of each blade 21 forms an angle a with the axial direction of the horizontal tank 2. Preferably, the included angle a is less than or equal to 20 degrees. As shown in fig. 5, the blades 21 are obviously not flat with the central axis of the tank, but have a certain included angle a, i.e. are arranged obliquely to the axis, so that the reaction liquid in each divided area can conveniently flow from one end of the tank cavity to the other end, and the end of the blade 21 is communicated with the adjacent area, so that the reaction liquid can flow to the adjacent area, and the mixing efficiency is improved. The included angle a should not be too large, that is, the blade 21 should not deviate from the axis too much, otherwise the reaction liquid will be accumulated at one end of the tank cavity, which is not beneficial to spreading.
In one embodiment of the horizontal self-stirring in vitro protein synthesis reactor according to the utility model, the profile of each of said blades 21 has an angle b <90 ° with the tangent plane of the horizontal tank 2 passing through the inner edge of said blade. It is explained that the paddle 21 is not completely vertical to the tank wall, but deviates from the vertical plane by a certain angle b, so that a certain downward pressure or lifting force can be generated on the reaction liquid, and the stirring effect can be improved according to the rotation direction of the tank body. Thereby obtaining more sufficient mixing.
In one embodiment of the horizontal self-stirring in vitro protein synthesis reactor of the present invention, the blades 21 are provided with a plurality of through holes (not shown) on the contour surface. Thus, the reaction liquids in the divided regions can interact with each other through the through holes, and the mixing effect is improved.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the present invention, the total area of the plurality of through holes on each of the blades 21 accounts for 30% to 50% of the area of the blade 21, and the distance between each adjacent through hole is equal. Thus, equal and equal amount of liquid flows in forward and reverse directions can be formed approximately, and the mixing effect is improved.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the present invention, the edge of each of the paddles 21 and the edge of the orifice of each of the through holes have a rounded structure. Such rounded corners may reduce shear forces on the active material from the blades 21.
In one embodiment of the horizontal self-stirring in vitro protein synthesis reactor of the present invention, the volume of the horizontal tank 2 is 10L-10000L. Along with the change of the volume of the tank body, the self-stirring blade assembly 20 in the tank body can also change correspondingly, such as increasing and decreasing the number of the blades 21 in the tank body, so as to adapt to different working conditions. The specific size of the volume of the tank body can be customized according to the capacity, and the volume of industrial mass production is preferably 1000L.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the present invention, as shown in fig. 7, a temperature-adjustable heating device 37 is disposed on the air inlet channel to adjust the temperature of the air entering the horizontal tank 2. Different reaction liquids have different optimal reaction temperatures, and the air inlet temperature can be adjusted according to actual needs so as to improve the reaction efficiency.
In one embodiment of the horizontal self-stirring in vitro protein synthesis reactor of the present invention, the air supply device is an air compressor (not shown) or a fan 8. When the air compressor is used, the output air pressure is adjustable. When the fan is a fan 8, the output air quantity is adjustable. Therefore, the air content in the tank cavity can be effectively controlled, different reaction liquid requirements are met, and the flexibility is improved.
In one embodiment of the horizontal self-stirring in vitro protein synthesis reactor of the present invention, the moving part is a rotary joint, the gas inlet channel is communicated with the horizontal tank 2 through a gas inlet rotary joint 32, and the gas outlet channel is communicated with the horizontal tank 2 through a gas outlet rotary joint 34. The rotary joint can realize the connection with the horizontal tank body 2 in the rotating process, so that the static part of the air passage can work normally, the external connection problem of the rotary tank body is effectively solved, and the replacement of the horizontal tank with the vertical tank becomes possible.
In one embodiment of the horizontal self-stirring in vitro protein synthesis reactor of the present invention, the tank cover assembly 3 is of an integral structure and is installed at the feed inlet of the horizontal tank body 2. As shown in fig. 1, 2, 4 and 6, the can lid assembly 3 is an integral unit and is installed at the feed opening of the left end of the horizontal can body 2. Therefore, the space of the whole set of equipment can be saved, and the installation and the use in a small space are convenient. And an opening at the other end of the tank body is also omitted, so that the processing cost is reduced.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the utility model, the tank cover assembly comprises a cover body 30 and a gas distribution device penetrating through the center of the cover body 30, the gas distribution device comprises a gas inlet pipe 31 and a gas outlet pipe 33, the gas inlet pipe 31 extends into the horizontal tank body 2, and the gas outlet pipe 33 is communicated with the horizontal tank body 2 through a gas outlet hole 301 formed in the cover body 30. Preferably, the discharge hole 301 has a plurality and surrounds the intake pipe 31. As shown in fig. 6, the air inlet pipe 31 and the exhaust pipe 33 are nested, and the air inlet pipe 31 on the left side of the cover 30 is sealed, so that the exhaust gas exhausted from the exhaust hole 301 on the cover 30 can not enter the air inlet pipe 31, the air inlet and outlet channels are effectively isolated, the structure is exquisite, and the air inlet and outlet effects are good.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the present invention, as shown in fig. 6, one end of the air inlet pipe 31 extending into the horizontal tank 2 is closed, and a plurality of air outlets are opened on the pipe wall. Preferably, the air outlet holes are uniformly distributed along the circumferential direction of the air inlet pipe 31 and extend along the axial direction of the air inlet pipe 31. Therefore, the air can be in full contact with the reaction liquid in the circumferential direction of the tank wall within the range of 360 degrees, and the reaction efficiency is improved.
In one embodiment of the horizontal self-stirring in vitro protein synthesis reactor of the present invention, as shown in fig. 4, the length of the gas inlet pipe 31 extending into the interior of the horizontal tank 2 is equivalent to the length of the horizontal tank 2. The air introduced in the way can be in full contact with the reaction liquid in the axial direction of the tank wall, and the reaction efficiency is improved.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the present invention, as shown in fig. 7, the tank cover assembly 3 is a split structure, and the tank cover assembly 3 includes an air inlet part and an air outlet part, the air inlet part is installed at the left end of the horizontal tank body 2, and the air outlet part is installed at the right end of the horizontal tank body 2. The structure with separated air inlet and outlet channels reduces the complexity of structural design and improves the working reliability. And the exhaust passage is conveniently communicated with an external exhaust gas treatment device.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the utility model, as shown in fig. 7, the air inlet part comprises an air inlet pipe 31, and the air inlet pipe 31 extends into the horizontal tank body 2; the exhaust part comprises a cover body and an exhaust pipe 33, and the exhaust pipe 33 is communicated with the horizontal tank body 2 through an exhaust hole formed in the cover body.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the present invention, as shown in fig. 2 and 3, the lid assembly 3 is provided with a sealing ring (not shown), a lid lock 35 and a locking handle 36, the horizontal tank 2 is provided with a tank lock 22, and the locking handle 36 is adapted to lock the lid lock 36 to the tank lock 22 so as to seal the lid 30 to the mouth, such as the feed inlet, of the horizontal tank 2. A ring of raised rings on the inside of the lid 30 can be fitted with a sealing ring to provide a better seal when the lid is closed to the can mouth.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the utility model, one end of the horizontal tank body 2 is supported by a bearing seat 5 arranged on the frame 1, the other end of the horizontal tank body is supported by a pair of rotating wheels 6 arranged on the frame 1, and the rotating wheels 6 are in contact with a circle of rolling ribs 23 on the outer wall of the horizontal tank body 2, so that when the horizontal tank body 2 rotates, the rolling ribs 23 roll along the pair of rotating wheels 6, and the rolling ribs protrude out of the outer wall of the tank body by a certain thickness, so that the horizontal tank body 2 can be protected.
In one embodiment of the horizontal self-stirring in-vitro protein synthesis reactor of the present invention, as shown in fig. 1, the driving device 4 is a motor, and the motor drives the horizontal tank 2 through a pulley transmission mechanism 7 or a gear transmission mechanism. The belt wheel transmission mechanism 7 can be adopted for the horizontal tank body 2 with small volume, and the gear transmission mechanism can be adopted for the horizontal tank body 2 with large volume.
In one embodiment of the horizontal self-stirring in vitro protein synthesis reactor according to the utility model, the axial direction of the horizontal tank 2 is parallel to the horizontal plane. The fully horizontal tank body structure can realize maximally spreading the reaction liquid in the tank body structure, prevent the reaction liquid from depositing to a certain direction and improve the reaction efficiency.
It should be noted that "horizontal" includes completely horizontal, i.e. the axial direction of the horizontal tank 2 is parallel to the horizontal plane, and also includes partially horizontal, or inclined, i.e. the axial direction of the horizontal tank 2 forms a certain angle with the horizontal plane. Which form is adopted can be completely adjusted according to actual needs.
When the horizontal self-stirring in-vitro protein synthesis reactor in any embodiment of the horizontal self-stirring in-vitro protein synthesis reactor is used for in-vitro protein synthesis reaction, the method comprises the following steps: (1) introducing a protein synthesis reaction solution which accounts for 5-30% of the volume of the horizontal tank body 2 into the horizontal tank body 2, and sealing; (2) and starting the driving device 4 to rotate the horizontal tank body 2 at a preset rotating speed r for a preset time T. The smaller the volume ratio of the protein synthesis reaction solution in the horizontal tank 2, the thinner the reaction solution is during the rotation of the tank, the higher the reaction efficiency, but the lower the yield, and a certain balance between the yield and the efficiency or activity is required. Preferably, in the step (2), while the driving device 4 is started, the air source device is started, air with a preset air pressure p is continuously input into the horizontal tank body 2 through the air inlet pipe 31, and waste gas generated in the reaction process is discharged through the air outlet pipe 33. More preferably, in step (2), r is 20rpm to 200rpm, T is 1h to 10h, and p is 10kPa to 300 kPa.
The direction of rotation of the horizontal tank 2 can be adjusted by controlling the forward and reverse rotation of the driving device 4, and the effect of mixing the reaction solution can be improved by intermittently or periodically adjusting the direction of rotation of the horizontal tank 2. The mixing effect of the reaction liquid can be improved by combining the adjustment of the steering and the rotating speed according to the actual requirement, and the effects of improving the reaction efficiency and the activity of the product are finally achieved.
Fig. 8 shows the results of activity tests on the target protein EGFP produced by the horizontal self-stirring in vitro protein synthesis reactor of the present invention in 1h, 3h, and 6h, respectively, wherein the volume of the horizontal self-stirring in vitro protein synthesis reactor is 50L, the volume of the reaction solution is 10L, r is 50rpm, and p is 1 atm, and the control group is the target protein produced in a laboratory using a 24-well plate. As can be seen from FIG. 8, the activity of the target protein EGFP obtained by using the reactor of the utility model is equivalent to that of a control group under three reaction times, and the activity is well maintained.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the utility model, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (29)
1. A horizontal self-stirring in-vitro protein synthesis reactor is characterized by comprising a frame, a horizontal tank body, a tank cover assembly and a driving device; the horizontal tank body is rotatably arranged on the machine frame, the tank cover assembly is detachably sealed on at least one end part of the horizontal tank body, and the driving device is used for driving the horizontal tank body to rotate; the inner wall of the horizontal tank body is provided with a self-stirring paddle component, and the self-stirring paddle component is suitable for stirring the protein synthesis reaction liquid in the horizontal tank body in the rotating process of the horizontal tank body.
2. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 1, further comprising a gas source device, wherein the tank cover assembly is provided with a gas inlet channel and a gas outlet channel, and the gas inlet channel is communicated with a gas outlet of the gas source device; the tank cover assembly comprises a moving part which is suitable for rotating along with the rotation of the horizontal tank body.
3. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 1, wherein the height of the self-stirring paddle assembly protruding out of the inner wall of the tank body accounts for no more than 60% of the radius of the inner circle of the horizontal tank body.
4. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 1, wherein the self-stirring paddle assembly comprises a plurality of paddles, each of the paddles extends in the axial direction of the horizontal tank body, and each of the paddles is arranged at intervals in the circumferential direction of the inner circle of the horizontal tank body.
5. The horizontal self-stirring in vitro protein synthesis reactor according to claim 4, wherein each of the paddles is arranged at equal intervals.
6. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 4, wherein each blade is integrally formed and fixed on the inner wall of the horizontal tank body by welding, and the welding seam is polished.
7. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 4, wherein one side of each blade, which is far away from the inner wall of the horizontal tank body, is of a concave arc structure.
8. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 4, wherein an included angle a is formed between the length direction of each blade and the axial direction of the horizontal tank body.
9. The horizontal self-stirring in vitro protein synthesis reactor according to claim 8, wherein the included angle a is less than or equal to 20 °.
10. The horizontal self-stirring in vitro protein synthesis reactor according to claim 4, wherein the profile of each of the blades has an angle b <90 ° with the tangent plane of the horizontal tank passing through the inner edge of the blade.
11. The horizontal self-stirring in-vitro protein synthesis reactor according to any one of claims 4 to 10, wherein the contour surface of each blade is provided with a plurality of through holes.
12. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 11, wherein the total area of the through holes on each blade accounts for 30-50% of the area of the blade, and the distance between every two adjacent through holes is equal.
13. The horizontal self-stirring in vitro protein synthesis reactor according to claim 11, wherein the edge of each blade and the edge of the orifice of each through hole have a rounded corner structure.
14. The horizontal self-stirring in vitro protein synthesis reactor according to claim 1, wherein the volume of the horizontal tank body is 10L-10000L.
15. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 2, wherein a heating device with adjustable temperature is arranged on the air inlet channel so as to adjust the temperature of air entering the horizontal tank body.
16. The horizontal self-stirring in vitro protein synthesis reactor according to claim 2, wherein the air source device is an air compressor or a fan.
17. The horizontal self-stirring in vitro protein synthesis reactor according to claim 2, wherein the moving part is a rotary joint, the air inlet channel is communicated with the horizontal tank body through an air inlet rotary joint, and the air outlet channel is communicated with the horizontal tank body through an air outlet rotary joint.
18. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 2, wherein the tank cover assembly is of an integrated structure and is arranged at the feed inlet of the horizontal tank body.
19. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 18, wherein the tank cover assembly comprises a cover body and a gas distribution device penetrating through the center of the cover body, the gas distribution device comprises a gas inlet pipe and a gas outlet pipe, the gas inlet pipe extends into the horizontal tank body, and the gas outlet pipe is communicated with the horizontal tank body through a gas outlet hole formed in the cover body.
20. The horizontal self-stirring in vitro protein synthesis reactor according to claim 19, wherein the gas outlet hole is provided in plurality and surrounds the gas inlet pipe.
21. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 19, wherein one end of the air inlet pipe, which extends into the horizontal tank body, is closed, and a plurality of air outlets are formed in the pipe wall of the air inlet pipe.
22. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 21, wherein the air outlets are uniformly distributed along the circumferential direction of the air inlet pipe and extend along the axial direction of the air inlet pipe.
23. The horizontal self-stirring in vitro protein synthesis reactor according to claim 19, wherein the length of the air inlet pipe extending into the interior of the horizontal tank is equivalent to the length of the horizontal tank.
24. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 2, wherein the tank cover assembly is of a split structure and comprises an air inlet part and an air outlet part, the air inlet part is arranged at the left end of the horizontal tank body, and the air outlet part is arranged at the right end of the horizontal tank body.
25. The horizontal self-stirring in vitro protein synthesis reactor according to claim 24, wherein the air inlet part comprises an air inlet pipe, and the air inlet pipe extends into the horizontal tank body; the exhaust part comprises a cover body and an exhaust pipe, and the exhaust pipe is communicated with the horizontal tank body through an exhaust hole formed in the cover body.
26. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 2, wherein the tank cover assembly is provided with a sealing ring, a tank cover lock catch and a locking handle, the horizontal tank body is provided with a tank body lock catch, and the locking handle is suitable for locking the tank cover lock catch on the tank body lock catch.
27. The horizontal self-stirring in vitro protein synthesis reactor according to claim 1, wherein one end of the horizontal tank body is supported by a bearing seat mounted on the frame, and the other end is supported by a pair of rotating wheels mounted on the frame.
28. The horizontal self-stirring in-vitro protein synthesis reactor according to claim 1, wherein the driving device is a motor, and the motor drives the horizontal tank body through a belt wheel transmission mechanism or a gear transmission mechanism.
29. The horizontal self-stirring in vitro protein synthesis reactor according to claim 1, wherein the axial direction of the horizontal tank body is parallel to the horizontal plane.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111448968 | 2021-11-30 | ||
| CN2021114489687 | 2021-11-30 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN216964638U true CN216964638U (en) | 2022-07-15 |
Family
ID=80796822
Family Applications (8)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202220250487.9U Active CN217473468U (en) | 2021-11-30 | 2022-01-26 | Tank cover integrated type air inlet and exhaust device of horizontal self-stirring external protein synthesis reactor |
| CN202210093587.XA Pending CN114247415A (en) | 2021-11-30 | 2022-01-26 | Horizontal self-stirring in-vitro protein synthesis reactor and in-vitro protein synthesis method |
| CN202210095041.8A Pending CN114307940A (en) | 2021-11-30 | 2022-01-26 | Tank device of horizontal self-stirring external protein synthesis reactor |
| CN202210093573.8A Pending CN114471367A (en) | 2021-11-30 | 2022-01-26 | Tank cover integrated type air inlet and exhaust device of horizontal self-stirring external protein synthesis reactor |
| CN202220216939.1U Active CN216964638U (en) | 2021-11-30 | 2022-01-26 | Horizontal self-stirring in-vitro protein synthesis reactor |
| CN202220528915.XU Active CN219279864U (en) | 2021-11-30 | 2022-03-11 | Bioreactor and corresponding partition, reaction device and shell |
| CN202220528922.XU Active CN217910448U (en) | 2021-11-30 | 2022-03-11 | Reaction device for in vitro biosynthesis |
| CN202210239438.XA Pending CN114405455A (en) | 2021-11-30 | 2022-03-11 | Reaction device for in vitro biosynthesis and in vitro biosynthesis method |
Family Applications Before (4)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202220250487.9U Active CN217473468U (en) | 2021-11-30 | 2022-01-26 | Tank cover integrated type air inlet and exhaust device of horizontal self-stirring external protein synthesis reactor |
| CN202210093587.XA Pending CN114247415A (en) | 2021-11-30 | 2022-01-26 | Horizontal self-stirring in-vitro protein synthesis reactor and in-vitro protein synthesis method |
| CN202210095041.8A Pending CN114307940A (en) | 2021-11-30 | 2022-01-26 | Tank device of horizontal self-stirring external protein synthesis reactor |
| CN202210093573.8A Pending CN114471367A (en) | 2021-11-30 | 2022-01-26 | Tank cover integrated type air inlet and exhaust device of horizontal self-stirring external protein synthesis reactor |
Family Applications After (3)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202220528915.XU Active CN219279864U (en) | 2021-11-30 | 2022-03-11 | Bioreactor and corresponding partition, reaction device and shell |
| CN202220528922.XU Active CN217910448U (en) | 2021-11-30 | 2022-03-11 | Reaction device for in vitro biosynthesis |
| CN202210239438.XA Pending CN114405455A (en) | 2021-11-30 | 2022-03-11 | Reaction device for in vitro biosynthesis and in vitro biosynthesis method |
Country Status (1)
| Country | Link |
|---|---|
| CN (8) | CN217473468U (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023098697A2 (en) * | 2021-11-30 | 2023-06-08 | 康码（上海）生物科技有限公司 | In-vitro biosynthesis reaction apparatus and in-vitro biosynthesis method |
| CN115320903B (en) * | 2022-10-14 | 2023-02-03 | 溧阳市宇峰新材料有限公司 | Filling equipment for sealant and using method thereof |
-
2022
- 2022-01-26 CN CN202220250487.9U patent/CN217473468U/en active Active
- 2022-01-26 CN CN202210093587.XA patent/CN114247415A/en active Pending
- 2022-01-26 CN CN202210095041.8A patent/CN114307940A/en active Pending
- 2022-01-26 CN CN202210093573.8A patent/CN114471367A/en active Pending
- 2022-01-26 CN CN202220216939.1U patent/CN216964638U/en active Active
- 2022-03-11 CN CN202220528915.XU patent/CN219279864U/en active Active
- 2022-03-11 CN CN202220528922.XU patent/CN217910448U/en active Active
- 2022-03-11 CN CN202210239438.XA patent/CN114405455A/en active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| CN217473468U (en) | 2022-09-23 |
| CN114307940A (en) | 2022-04-12 |
| CN114405455A (en) | 2022-04-29 |
| CN114247415A (en) | 2022-03-29 |
| CN114471367A (en) | 2022-05-13 |
| CN219279864U (en) | 2023-06-30 |
| CN217910448U (en) | 2022-11-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN216964638U (en) | Horizontal self-stirring in-vitro protein synthesis reactor | |
| US8822208B2 (en) | Bioreactor and method | |
| US5075234A (en) | Fermentor/bioreactor systems having high aeration capacity | |
| AU2010238548B2 (en) | Cell culture system | |
| CN209005951U (en) | It is a kind of to realize highly purified quartz sand flotation machine | |
| CN1578830A (en) | A unit and a process for carrying out high cell density fermentation | |
| CN213624085U (en) | Biological fermentation device with multiple stirring shafts | |
| KR20140056173A (en) | Medium-stirring pulverizer | |
| CN114127248A (en) | Bioreactor for growing microorganisms | |
| WO2013188649A1 (en) | Pneumatically agitated and aerated single-use bioreactor | |
| CN106281979B (en) | High-efficiency vertical fermentation tank | |
| CN209113882U (en) | Herb fermenting device | |
| JPS633590B2 (en) | ||
| CN207567221U (en) | A kind of intelligent biological bacterial manure production parallel fermentation system of stalk solid-liquid | |
| CN107602172A (en) | Bio-fertilizer installation for fermenting | |
| EP3561046B1 (en) | Dynamic solid-state fermentation apparatus | |
| RU2610674C1 (en) | Bioreactor for biochemical processes | |
| CN107651981A (en) | Bio-feritlizer installation for fermenting with heating function | |
| RU2363729C1 (en) | Apparatus for suspension cultivation of tissue or microorganism cells | |
| CN210237605U (en) | Energy-saving fermentation tank | |
| CN209333755U (en) | A kind of solid-liquid hybrid reactor | |
| JP2004261659A (en) | Mixing apparatus | |
| CN218969125U (en) | Microbial fertilizer preparation fermentation tank | |
| CN218202766U (en) | High flux fermenting installation convenient to filamentous fungi collects | |
| CN212357229U (en) | Fermentation tank |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20231215 Address after: No. 2 Nanya Road, Minhang District, Shanghai, 201321 Patentee after: KANGMA-HEATHCODE (SHANGHAI) BIOTECH Co.,Ltd. Patentee after: Yanshimei Society (Hainan) Medical Beauty Health Technology Co.,Ltd. Address before: Building 12, Lane 118, Furonghua Road, Pudong New Area, Shanghai, 201321 Patentee before: KANGMA-HEATHCODE (SHANGHAI) BIOTECH Co.,Ltd. |
|
| TR01 | Transfer of patent right |