CN220737514U - Recombinant protein separation device - Google Patents
Recombinant protein separation device Download PDFInfo
- Publication number
- CN220737514U CN220737514U CN202322156464.9U CN202322156464U CN220737514U CN 220737514 U CN220737514 U CN 220737514U CN 202322156464 U CN202322156464 U CN 202322156464U CN 220737514 U CN220737514 U CN 220737514U
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- CN
- China
- Prior art keywords
- recombinant protein
- semipermeable membrane
- membrane filter
- filter disc
- box body
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- 102000007056 Recombinant Fusion Proteins Human genes 0.000 title claims abstract description 54
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 title claims abstract description 54
- 238000000926 separation method Methods 0.000 title claims abstract description 34
- 239000007788 liquid Substances 0.000 claims abstract description 34
- 239000012528 membrane Substances 0.000 claims abstract description 25
- 239000012266 salt solution Substances 0.000 claims abstract description 18
- 238000001556 precipitation Methods 0.000 claims abstract description 15
- 239000011550 stock solution Substances 0.000 claims abstract description 9
- 238000003756 stirring Methods 0.000 claims abstract description 4
- 238000005273 aeration Methods 0.000 claims description 10
- 238000007790 scraping Methods 0.000 claims description 8
- 239000002244 precipitate Substances 0.000 claims description 4
- 238000004062 sedimentation Methods 0.000 claims 1
- 102000004169 proteins and genes Human genes 0.000 abstract description 18
- 108090000623 proteins and genes Proteins 0.000 abstract description 18
- 238000006243 chemical reaction Methods 0.000 abstract description 13
- 238000000034 method Methods 0.000 abstract description 7
- 239000000243 solution Substances 0.000 abstract description 7
- 230000005611 electricity Effects 0.000 abstract description 5
- 238000001914 filtration Methods 0.000 abstract description 5
- 238000002156 mixing Methods 0.000 abstract description 2
- 238000005185 salting out Methods 0.000 description 11
- 239000000463 material Substances 0.000 description 7
- 150000003839 salts Chemical class 0.000 description 7
- 239000012460 protein solution Substances 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000000149 penetrating effect Effects 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000006920 protein precipitation Effects 0.000 description 1
- -1 salt ions Chemical class 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Landscapes
- Separation Using Semi-Permeable Membranes (AREA)
- Peptides Or Proteins (AREA)
Abstract
The utility model discloses a recombinant protein separation device which comprises a liquid storage box body, wherein one side of the liquid storage box body is connected with a liquid discharge valve in a threaded manner, a reaction kettle is fixedly welded at the top end of the liquid storage box body, a precipitation mechanism is arranged in the liquid storage box body and used for carrying out precipitation filtration on recombinant proteins in separated clear liquid, so that the situation that fine proteins are mixed in the clear liquid is avoided, and a separation mechanism is arranged in the reaction kettle and used for carrying out ultrasonic stirring on the interior of a salt solution and carrying out separation treatment on proteins in the stock solution. In the process of operation, drive the electricity storage layer board seat through ball and carry out the repeated lift processing, provide the power for the ultrasonic module in outside through the electricity storage layer board seat, carry out ultrasonic wave mixing repeatedly to equipment inside through ultrasonic module, conveniently carry out the vertical reaction to the inside of jar body to utilize the semipermeable membrane filter disc of ball drive bottom to rotate, drive recombinant protein's clear solution and carry out rotatory filtration.
Description
Technical Field
The utility model relates to the technical field of protein separation, in particular to a recombinant protein separation device.
Background
The recombinant protein is produced by using recombinant DNA or recombinant RNA technology, and after the recombinant protein is produced, the recombinant protein is usually required to be taken out of a host cell, at this time, the recombinant protein and the thallus hybrid protein are required to be separated, and when the recombinant protein is separated, a common mode is a salting-out method, namely, the recombinant protein is separated out successively by utilizing different solubilities of different proteins in saline water, so as to fulfill the aim of separation.
However, the existing recombinant protein separation device only depends on a salting-out method, and has the defects of long protein separation time and low separation efficiency in the operation process, and the recombinant protein separation device mainly performs separation treatment on the stock solution through an aeration structure or a catalytic mechanism in the operation process, but has a single mixed structure, and needs aeration or a mixed structure to mix the stock solution, so that frequent reaction separation is performed in the salt solution repeatedly, but the recombinant protein is easy to denature in the long-time separation process, or the recombinant protein has low yield, low purity and the like.
Disclosure of Invention
The present utility model is directed to a recombinant protein separation apparatus that overcomes the above-mentioned drawbacks of the prior art.
The utility model provides a recombinant protein separator, includes the liquid reserve tank body, one side threaded connection of liquid reserve tank body has the flowing back valve, the top welded fastening of liquid reserve tank body has reation kettle, the inside of liquid reserve tank body is provided with precipitation mechanism, precipitation mechanism carries out precipitation to the inside recombinant protein of clear liquid of separation and filters, avoids slight recombinant protein to mix inside the clear liquid, reation kettle's inside is provided with separating mechanism, separating mechanism carries out the stirring of ultrasonic wave to the salt solution inside, carries out separation treatment to recombinant protein in the stoste.
Preferably, the precipitation mechanism comprises a semipermeable membrane filter disc, a rotating shaft, an electromagnetic valve and a scraping sheet, wherein the top end of the semipermeable membrane filter disc is connected with the rotating shaft in a key manner, the scraping sheet is arranged at the bottom end of the semipermeable membrane filter disc, the semipermeable membrane filter disc is arranged inside a liquid storage box body, and the electromagnetic valve is connected with the top end of the liquid storage box body in a threaded manner.
Preferably, the semipermeable membrane filter disc is connected with the top end of the ball screw in a nested way through a rotating shaft at the top end of the semipermeable membrane filter disc.
Preferably, the separating mechanism comprises a guide pipeline, an electric storage supporting plate seat, an ultrasonic module, a ball screw and an aeration hole, wherein the electric storage supporting plate seat is connected to the outer side of the guide pipeline in a penetrating manner, the ultrasonic module is connected to an electric wire on the outer side of the electric storage supporting plate seat, the ball screw is connected to the outer side of the electric storage supporting plate seat in a threaded manner, and the aeration hole is formed in the outer side of the guide pipeline at equal intervals.
Preferably, the electric storage supporting plate seat is connected with the output end of the driving motor through a ball screw arranged on the outer side of the electric storage supporting plate seat.
Compared with the prior art, the utility model has the following advantages:
1. when the salt concentration in the solution is increased to a certain concentration, the solubility of the protein is reduced to different degrees and the protein is separated out successively, the phenomenon is called salting out, and the salt saturation required by salting out of different proteins is different, SO that the target recombinant protein can be precipitated out by adjusting the salt concentration, the moving range of an ultrasonic module is dynamically adjusted, the inside of the salt solution is vertically lifted and mixed, the efficiency of separating the recombinant protein by salting out is accelerated, the recombinant protein solution is poured into the reaction kettle from a feed inlet when the solution is used, a large amount of salt is added into the reaction kettle from a salt inlet box, and high-concentration salt ions (such as SO and NH of ammonium sulfate) have strong hydration power.
2. In the process of operation, drive the electric power storage layer board seat through ball and carry out the processing of lifting repeatedly, the ultrasonic module that is the outside through the electric power storage layer board seat provides the power, carry out ultrasonic wave mixing repeatedly through ultrasonic wave module to equipment inside, the convenience carries out vertical reaction to the inside of jar body and handle, and utilize ball to drive the semipermeable membrane filter disc of bottom and rotate, drive recombinant protein's clear solution and carry out rotatory filtration, reduce the residual content of recombinant protein after the separation of clear solution inside, recombinant protein's salt saturation is very high, then precipitate thallus miscellaneous protein earlier, recombinant protein remains in the protein solution, if recombinant protein salt saturation is lower, then precipitate it together with partial miscellaneous protein.
Drawings
FIG. 1 is a schematic view of the overall three-dimensional structure of the present utility model.
FIG. 2 is a schematic view of the structure of the inside of the reaction vessel in the present utility model.
Fig. 3 is a schematic structural diagram of the liquid storage tank body according to the present utility model.
Wherein:
1. a liquid storage tank body; 2. a liquid discharge valve; 3. a reaction kettle; 4. a guide duct; 5. a driving motor; 6. a precipitation mechanism; 7. a separation mechanism; 8. an electric storage pallet seat; 9. an ultrasonic module; 10. a ball screw; 11. aeration holes; 12. a semipermeable membrane filtration disc; 13. a rotation shaft; 14. an electromagnetic valve; 15. scraping the material sheet.
Detailed Description
The utility model is further described in connection with the following detailed description, in order to make the technical means, the creation characteristics, the achievement of the purpose and the effect of the utility model easy to understand.
As shown in fig. 1 to 3, a recombinant protein separation device comprises a liquid storage box body 1, one side threaded connection of liquid storage box body 1 has flowing back valve 2, the top welded fastening of liquid storage box body 1 has reation kettle 3, the inside of liquid storage box body 1 is provided with precipitation mechanism 6, precipitation mechanism 6 carries out precipitation to the inside recombinant protein of clear liquid of separation and filters, avoids fine protein to mix inside the clear liquid, the inside of reation kettle 3 is provided with separating mechanism 7, separating mechanism 7 carries out ultrasonic stirring to the inside of salt solution and separation recombinant protein, carries out separation treatment to the protein in the stoste.
In this embodiment, the precipitation mechanism 6 includes a semipermeable membrane filter disc 12, a rotation shaft 13, an electromagnetic valve 14 and a scraping plate 15, the top end of the semipermeable membrane filter disc 12 is connected with the rotation shaft 13 in a key manner, the scraping plate 15 is disposed at the bottom end of the semipermeable membrane filter disc 12, the semipermeable membrane filter disc 12 is disposed inside the liquid storage tank 1, the electromagnetic valve 14 is screwed to the top end of the liquid storage tank 1, and the separated salt solution and recombinant protein are directly introduced into the precipitation mechanism 6 through the electromagnetic valve 14.
In this embodiment, the semipermeable membrane filter disc 12 is nested and connected with the top end of the ball screw 10 through the rotating shaft 13 at the top end of the semipermeable membrane filter disc 12, and when the semipermeable membrane filter disc 12 is rotating and filtering, the ball screw 10 drives the bottom ultrasonic module 9 to repeatedly lift and lower.
In this embodiment, the separating mechanism 7 includes a guide pipe 4, an electric storage support plate seat 8, an ultrasonic module 9, a ball screw 10 and an aeration hole 11, the outer side of the guide pipe 4 is connected with the electric storage support plate seat 8 in a penetrating manner, an outer side wire of the electric storage support plate seat 8 is connected with the ultrasonic module 9, an outer side thread of the electric storage support plate seat 8 is connected with the ball screw 10, the outer side of the guide pipe 4 is provided with the aeration hole 11 at equal intervals, and gas is continuously introduced into the salt solution through the guide pipe 4, so that the efficiency of material catalytic reaction inside the salt solution is accelerated.
In this embodiment, the electricity storage pallet seat 8 is connected to the output end of the driving motor 5 through a ball screw 10 provided outside the electricity storage pallet seat 8, and the material separation uniformity is accelerated by repeatedly lifting and lowering the electricity storage pallet seat 8 in the reaction kettle 3.
The recombinant protein separation device comprises the following working contents in practical application:
step 1: an operator firstly adds a recombinant protein solution to be separated into a salting-out kettle, and separates out the recombinant protein by dehydration under the salting-out action of the salt solution, and simultaneously, the recombinant protein is directly led into the reaction kettle 3 while being separated out, and the salt solution for separating the recombinant protein is led into the reaction kettle 3, the operator turns on a driving motor 5, drives a ball screw 10 to rotate by the driving motor 5, drives a separating mechanism 7 to rotate and regulate by the ball screw 10, and vertically descends the outer side of the separating mechanism 7 by a guide pipeline 4, and utilizes an internal electric storage supporting plate seat 8 of the separating mechanism 7 to provide a power supply for an ultrasonic module 9, and the salt solution and the recombinant protein stock solution are mixed in a vibrating way by the ultrasonic module 9, so that ultrasonic waves are emitted into the solution by the ultrasonic module 9, and cell rupture can be further accelerated by the effect of the ultrasonic waves, and protein substances are exposed;
step 2: the amplitude of the ultrasonic module 9 can be executed for 24 hours at 20-40%, meanwhile, the guide pipeline 4 is connected with the gas pipeline, gas is continuously introduced into the salt solution through the aeration holes 11, the material separation speed in the salt solution is accelerated, the oxidation speed in the material is accelerated, and the protein is separated from the stock solution;
step 3: then opening the electromagnetic valve 14, leading out the salt solution in the reaction kettle 3 and the reacted recombinant protein precipitation solution by the electromagnetic valve 14, directly discharging the salt solution, leading the separated salt solution into the semipermeable membrane filter disc 12, simultaneously driving the rotating shaft 13 and the semipermeable membrane filter disc 12 to rotate by using the driving motor 5, adding NaCl into the supernatant in a mode of being 5wt%, thereby obtaining white precipitation, and separating and filtering the protein and the supernatant in the reacted salt solution by using the semipermeable membrane filter disc 12;
step 4: after the white precipitate is centrifugally separated and dialyzed into refined water, freeze drying is carried out to extract recombinant protein, an operator can scrape the precipitated material in the liquid storage box body 1 by using a scraping plate 15 at the bottom end of a semipermeable membrane filter disc 12 to avoid the adhesion of the material to the bottom end of the liquid storage box body 1, the operator opens a liquid discharge valve 2 to directly discharge the reacted clear liquid and separated protein, only the recombinant protein is in a protein solution for dialysis, the protein solution carrying the recombinant protein is poured into a salting-out device for secondary salting-out after the dialysis is finished, and if the recombinant protein is in a protein colloidal particle, the recombinant protein is poured into the salting-out device for secondary salting-out after being dissolved, and the process is repeated until the recombinant protein with high purity is obtained.
Accordingly, the above disclosed embodiments are illustrative in all respects, and not exclusive. All changes that come within the scope of the utility model or equivalents thereto are intended to be embraced therein.
Claims (5)
1. A recombinant protein separation device, characterized in that: including stock solution box (1), one side threaded connection of stock solution box (1) has flowing back valve (2), the top welded fastening of stock solution box (1) has reation kettle (3), the inside of stock solution box (1) is provided with precipitation mechanism (6), precipitation mechanism (6) precipitate the inside recombinant protein of clear liquid of separation and filter, avoid slight recombinant protein to mix inside the clear liquid, the inside of reation kettle (3) is provided with separating mechanism (7), separating mechanism (7) carry out the stirring of ultrasonic wave to the salt solution inside, carry out separation treatment to recombinant protein in the stoste.
2. A recombinant protein separation apparatus according to claim 1, wherein: the sedimentation mechanism (6) comprises a semipermeable membrane filter disc (12), a rotating shaft (13), an electromagnetic valve (14) and a scraping sheet (15), wherein the top end of the semipermeable membrane filter disc (12) is connected with the rotating shaft (13) through a key, the scraping sheet (15) is arranged at the bottom end of the semipermeable membrane filter disc (12), the semipermeable membrane filter disc (12) is arranged inside a liquid storage box body (1), and the electromagnetic valve (14) is connected with the top end of the liquid storage box body (1) through threads.
3. A recombinant protein separation apparatus according to claim 2, wherein: the semipermeable membrane filter disc (12) is connected with the top end of the ball screw (10) in a nested way through a rotating shaft (13) at the top end of the semipermeable membrane filter disc (12).
4. A recombinant protein separation apparatus according to claim 1, wherein: separating mechanism (7) are including direction pipeline (4), electric power storage layer board seat (8), ultrasonic module (9), ball (10) and aeration hole (11), the outside through connection of direction pipeline (4) has electric power storage layer board seat (8), the outside wire connection of electric power storage layer board seat (8) has ultrasonic module (9), the outside threaded connection of electric power storage layer board seat (8) has ball (10), aeration hole (11) have been seted up to equidistant outside of direction pipeline (4).
5. The recombinant protein separation apparatus according to claim 4, wherein: the electric storage supporting plate seat (8) is connected with the output end of the driving motor (5) through a ball screw (10) arranged on the outer side of the electric storage supporting plate seat (8).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202322156464.9U CN220737514U (en) | 2023-08-11 | 2023-08-11 | Recombinant protein separation device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202322156464.9U CN220737514U (en) | 2023-08-11 | 2023-08-11 | Recombinant protein separation device |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN220737514U true CN220737514U (en) | 2024-04-09 |
Family
ID=90551367
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202322156464.9U Active CN220737514U (en) | 2023-08-11 | 2023-08-11 | Recombinant protein separation device |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN220737514U (en) |
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2023
- 2023-08-11 CN CN202322156464.9U patent/CN220737514U/en active Active
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