CN207958195U - A kind of protein purification device - Google Patents
A kind of protein purification device Download PDFInfo
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- CN207958195U CN207958195U CN201721861786.1U CN201721861786U CN207958195U CN 207958195 U CN207958195 U CN 207958195U CN 201721861786 U CN201721861786 U CN 201721861786U CN 207958195 U CN207958195 U CN 207958195U
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Abstract
The utility model discloses a kind of protein purification device, including sequentially connected chromatographic column, pH regulating devices, the pH regulating devices are sequentially connected dialysis cartridge, purifying plant, collecting tank, the purifying plant includes purifying slot, it is respectively set in the both sides of the purifying slot and is connected to generate electric field with the high voltage power supply in control system by conducting wire, switch with the positive and negative pole plate of the metal of external insulation, the positive and negative pole plate of metal.A kind of protein purification device described in the utility model carries out smart purification after slightly being purified to target protein, the target protein purity purified is high.
Description
Technical field
The utility model belongs to equipment for purifying, and in particular to a kind of protein purification device.
Background technology
For protein using extensively in biological, chemistry, medicament research application, protein is answered in isolating and purifying for protein
Apparatus has a major impact, and is present in due to protein in complicated biosystem, and itself is unstable, heat or the certain solvents of chance
It is easy deactivation, therefore the isolation technics such as traditional distillation, solvent extraction are not particularly suited for isolating and purifying for protein.Currently,
As long as the isolation and purification method of protein includes following several：
1, supercentrifugation
The sinking speed in gradient liquid is different using various particles for this method, makes to have the particle of different sinking speed to be in not
With in density gradient layer, achieving the purpose that separation and protein purification, therefore ultracentrifugation or gradient density centrifugal isolate and purify
Method is only applicable to the separation of a small number of macro-molecular proteins, such as IgM, C1q, thyroglobulin etc., in most, small-molecular-weight egg
It is difficult purifying that white matter, which adopts this method,.
2, selective precipitation
This method promotes albumen according to the difference of each protein physicochemical property using various precipitating reagents or the certain conditions of change
Matter antigenic component precipitates, to achieve the purpose that purifying, most common method are salt precipitation methods.The principle of salting out method is albumen
The solubility of matter in aqueous solution depends on the hydrone number around protein molecule surface ion, that is, mainly by albumen
What the case where matter molecule periphery hydrophilic radical and the degree and protein molecule of water formation hydration shell carry charge determined.Albumen
After neutral salt is added in matter solution, since the affinity of neutral salt and hydrone is more than protein, cause around protein molecule
Hydrated sheath weaken so that disappear.Meanwhile neutral salt is added after protein solution since ionic strength changes, protein table
The charge in face is largely neutralized, and more protein solubility is caused to reduce, and makes to assemble and precipitate between protein molecule.Due to each
The protein of different solubility of the kind protein in different salinity, the salting liquid precipitation of different saturation is different, to make
Come out from other Protein Separations.Most common salting liquid is the ammonium sulfate of 33%~50% saturation degree.Salting out method is simple and convenient,
It can be used for the thick of proteantigen to carry, the extraction of gamma globulin, the concentration etc. of protein.The antigen purity of salting out method purification is not
Height is only applicable in the preliminary purification of antigen.
3, gel chromatography
Gel chromatography is detached to protein using molecular sieve effect.Gel is with three dimensions holey knot
The substance of structure is packed into chromatographic column after solution equilibria appropriate.A kind of sample solution containing various molecules slowly flows through
When chromatographic column, macromolecular substances are not easily accessible the micropore of gel particle, can only be distributed between particle, therefore downward in elution
Mobile speed, is eluted at first.Small-molecule substance can also pass into solidifying other than it can be spread in gel particle gap
In the micropore of glue particle, the speed that when elution moves down is slower, is then eluted.Therefore, protein molecule presses molecular size
It is detached, therefore is only applicable to the thick step purifying of protein using this method.
4, affinity chromatography
Affinity chromatography is the biologic specificity using large biological molecule, i.e., there is single-minded affinity between large biological molecule and
The chromatographic technique of design.Such as antigen and antibody, enzyme and enzyme inhibitor (or ligand), zymoprotein and coenzyme, hormone and receptor,
There is a kind of special affinity between the substances such as IgG and staphylococcal protein A (SPA).Such as when purification IgG, SPA can be adsorbed
On an inert solid-phase matrix, and it is prepared into chromatographic column.When sample flows through chromatographic column, IgG to be separated can be sent out with SPA
Raw specific binding, remaining ingredient cannot be in combination.After chromatographic column is fully eluted, change the ionic strength or pH of eluent
Value, IgG are dissociated with the SPA on solid-phase matrix, and the IgG to be purified can be obtained by collecting eluent.Affinity chromatography purifying protein
The major advantage of matter antigen is purity height, simple and fast, but cost is more expensive.
Invention content
The purpose of this utility model is in view of the problems of the existing technology, to provide a kind of protein purification device, described
Protein purification device being capable of efficiently purifying multiple proteins.
To achieve the above object, the technical solution adopted in the utility model is：
A kind of protein purification device, including sequentially connected chromatographic column, pH regulating devices, the pH regulating devices are successively
Dialysis cartridge, purifying plant, collecting tank are connected, the purifying plant includes purifying slot, is respectively set in the both sides of the purifying slot
With the positive and negative pole plate of metal of external insulation, the positive and negative pole plate of metal passes through conducting wire, switch and the height in the control system
Voltage source connects to generate electric field.
It is closed the switch between the positive and negative pole plate of the metal and the high voltage power supply by the control system, it is to be purified
Protein example carries out chromatography screening into the chromatographic column, and the protein after screening enters the pH regulating reservoirs, the pH tune
Target protein is adjusted to isoelectric point protein by the adjusting liquid in section pond, and the protein after balance enters in the dialysis cartridge,
Protein after dialysis enters in the purifying slot of the purifying plant, because target protein is isoelectric point protein, in electricity
It is not acted on by electric field force in, therefore, target protein falls along straight line under gravity, into collecting tank, and its
Remaining protein is that charged protein is deflected by electric field force effect, is enriched on positive and negative sheet metal pole plate.
To realize while purifying the protein of different molecular weight, the side wall setting one or more of the gel permeation column goes out
Liquid mouth, the liquid outlet are sequentially connected the pH regulating devices, dialysis cartridge, purifying plant, collecting tank.
For ease of the chromatographic column is added in protein example to be purified, the top of the chromatographic column is connected with sample addition zone.
For convenience of being balanced to gel layer in the chromatographic column, the chromatography top end is connected with the compensator, institute
The bottom for stating chromatographic column is connected with waste liquid tank, and equilibrium liquid is stored in the compensator, and equilibrium liquid is flowed by chromatographic column top,
Bottom is flowed out, and waste liquid tank is eventually entered into.
Preferably, to recycle equilibrium liquid, pass through the bottom of circulation conduit and the waste liquid tank at the top of the compensator
It is connected, circulating pump is set on the circulation conduit, the circulating pump is electrically connected with control system, and institute is opened by the control system
Circulating pump is stated, the equilibrium liquid in the compensator flows at the top of the chromatographic column, bottom outflow, into the waste liquid tank, institute
It states circulating pump and the waste liquid in the waste liquid tank is pumped into the compensator, it is flat to carry out cycle to the gel layer in the chromatographic column
Weighing apparatus.
It is furthermore preferred that a solenoid valve is arranged between the waste liquid tank, circulating pump, the solenoid valve is electrically connected with control system.
For ease of controlling the pH in the pH regulating reservoirs, the pH regulating devices include liquid-adding cell, pH regulating reservoirs and pH tune
The connected pH measuring systems in pond are saved, the pH regulating reservoirs are connected by a solenoid valve with the liquid-adding cell bottom, and the pH is measured
System is electrically connected with the control system.Protein after screening enters the pH regulating reservoirs, and the pH-meter measures the pH
PH in regulating reservoir, the control system pass through the electromagnetism valve regulation institute between the pH regulating reservoirs, liquid-adding cell according to measurement data
The flow velocity that the buffer solution in liquid-adding cell flows into the pH regulating reservoirs is stated, the adjusting liquid in the pH regulating reservoirs is by target protein
It is adjusted to isoelectric point protein.
Preferably, in the dialysis cartridge setting only allow protein by semi-permeable membrane, the semi-permeable membrane is by the dialysis
Column is divided into solution storage chamber, protein storage room, and the solution storage chamber is connected with holding vessel, the protein storage room and institute
The top for stating purifying plant is connected, and the bottom of the purifying plant is connected with the collecting tank.
Preferably, for ease of controlling the flowing velocity of the protein example, the sample addition zone and chromatographic column, chromatographic column with
It is respectively provided with solenoid valve between pH regulating devices, pH regulating reservoirs and liquid-adding cell, pH regulating reservoirs and dialysis cartridge, purifying plant and collecting tank.
The solenoid valve is electrically connected with the control system.
For convenience of the pump speed of peristaltic pump between the chromatographic column, regulating device is arranged, protein compression is set in the chromatographic column
Measuring appliance is spent, the protein concentration measuring appliance is electrically connected with control system, and the protein concentration measuring appliance measures the chromatography
The concentration of target protein and the control system is transferred data in column, when the concentration of target protein reaches setting value
Afterwards, the wriggling pump work between the chromatographic column, regulating device, the control system are arranged automatically according to the concentration of target protein
The pump speed of peristaltic pump between the chromatographic column, regulating device.
To improve the purification rate of protein, the pH regulating devices further include the protein being connected with the pH regulating reservoirs
Measurement of concetration device, the protein concentration measuring appliance are electrically connected with the control system, and the control system is according in pH regulating reservoirs
Protein concentration be arranged the pH regulating devices, dialyse intercolumniation peristaltic pump pump speed.
For convenience of the pH value adjusted in the pH regulating reservoirs, the pH regulating devices include liquid-adding cell, pH regulating reservoirs and institute
The connected pH measuring systems of pH regulating reservoirs are stated, the pH measuring systems are electrically connected with the control system, and the pH regulating reservoirs pass through
One solenoid valve is connected with the liquid-adding cell bottom, the control system according to pH value in pH regulating reservoirs be arranged the pH regulating reservoirs,
The relevant parameter of solenoid valve between liquid-adding cell adjusts pH adjustings liquid stream in liquid-adding cell and enters the speed in the pH regulating reservoirs.
It is described to control the display panel control institute to realize that visualized operation, the control system include controlling the display panel
The relevant parameter of the solenoid valve, peristaltic pump is arranged in the opening and closing for stating solenoid valve, peristaltic pump, shows the pH measuring systems, albumen
The measured value of matter measurement of concetration device.
Compared with prior art, the utility model has the beneficial effects that：
1, a kind of protein purification device described in the utility model includes：Chromatographic column, pH regulating devices, dialysis cartridge, purification
Device, for target protein by thick purification, essence purification, the target protein purity purified is high.
2, a kind of protein purification device described in the utility model can recycle equilibrium liquid, improve the utilization of equilibrium liquid
Rate reduces the pollution to environment, cost-effective.
3, a kind of protein purification device described in the utility model is totally-enclosedization pipeline purifying plant, avoids protein quilt
Air pollution keeps the activity of protein.
4, a kind of protein purification apparatus structure described in the utility model is simple and convenient to operate, work efficiency is high, can be a large amount of
Purify target protein.
Description of the drawings
Fig. 1 is a kind of structural schematic diagram of protein purification device embodiment 1 described in the utility model；
Fig. 2 is a kind of structural schematic diagram of protein purification device embodiment 2 described in the utility model；
In figure：1, chromatographic column；101, waste liquid tank；103, compensator；104, circulating pump；105, circulation conduit；106, it is loaded
Room；2, pH regulating reservoirs；201, liquid-adding cell；3, dialysis cartridge；301, semi-permeable membrane；303, holding vessel；4, purifying plant；5, collecting tank；
6, peristaltic pump；7, solenoid valve.
Specific implementation mode
Below in conjunction with the attached drawing in the utility model, the technical solution of the utility model is clearly and completely retouched
It states, it is clear that the described embodiments are only a part of the embodiments of the utility model, instead of all the embodiments.Based on this
Embodiment in utility model, those of ordinary skill in the art obtained under the conditions of not making creative work it is all its
Its embodiment, shall fall within the protection scope of the present invention.
Embodiment 1
As shown in Figure 1, a kind of protein purification device, including sequentially connected chromatographic column 1, pH regulating reservoirs 2, dialysis cartridge 3,
Purifying plant 4, collecting tank 5.The top of the chromatographic column 1 is connected with sample addition zone 106, the pH regulating reservoirs 2 and the liquid-adding cell
201 bottoms, the pH measuring systems are connected, and are arranged semi-permeable membrane 301 in the dialysis cartridge 3, and the semi-permeable membrane 301 is by the dialysis
Column 3 divides for solution storage chamber, protein storage room, and the solution storage chamber and 303 top of the holding vessel are connected, the albumen
Matter storage room is connected with the top of the purifying plant 4, and the bottom of the purifying plant 4 is connected with 5 top of the collecting tank.Institute
It states and is respectively provided with peristaltic pump 6, institute between chromatographic column 1 and pH regulating reservoirs 2, pH regulating reservoirs 2 and dialysis cartridge 3, dialysis cartridge 3 and purifying plant 4
State sample addition zone 106 and chromatographic column 1, chromatographic column 1 and pH regulating reservoirs 2, pH regulating reservoirs 2 and liquid-adding cell 201, pH regulating reservoirs 2 and dialysis
It is respectively provided with solenoid valve 7 between column 3, purifying plant 4 and collecting tank 5.The purifying plant 4 includes purifying slot, in the purifying slot
Both sides are respectively provided with passes through conducting wire, switch and the control with the positive and negative pole plate of the metal of external insulation, the positive and negative pole plate of metal
High voltage power supply in system connects to generate electric field.The pH measuring systems, solenoid valve 7, peristaltic pump 6, switch and control system
It is electrically connected.
Protein example to be purified is put into sample addition zone 106, the peristaltic pump 6, electromagnetism are opened by control system
Valve 7, the switch being closed between the positive and negative pole plate of metal and the high voltage power supply, and set sample to be purified and instill the chromatography
Drop speed in column 1.Protein example in the sample addition zone 106 instills in the chromatographic column 1, and the chromatographic column 1 and pH is adjusted
Protein after screening is pumped into pH regulating reservoirs 2, the pH measuring systems, control system, pH regulating reservoirs 2 by the peristaltic pump 6 between pond 2
7 triple interaction of solenoid valve between liquid-adding cell 201 adjusts the pH in the pH regulating reservoirs 2.Tune in the pH regulating reservoirs 2
Target protein is adjusted to isoelectric point protein by section liquid, and the peristaltic pump 6 between the pH regulating reservoirs 2, dialysis cartridge 3 will be after balance
Protein is pumped into dialysis cartridge 3, and the protein enters protein storage room by the semi-permeable membrane 301 in the dialysis cartridge 3, is remained
Remaining solution stays in solution storage chamber and flows into the holding vessel 303, and the peristaltic pump 6 between the dialysis cartridge 3, purifying plant 4 is by institute
It states the protein in protein storage room to be pumped into the purifying slot of the purifying plant 4, because target protein is isoelectric point egg
White matter is not acted in the electric field by electric field force, and therefore, target protein falls along straight line under gravity, into institute
Collecting tank 5 is stated, and remaining protein is deflected for charged protein by electric field force effect, on positive and negative sheet metal pole plate
Enrichment.
A kind of protein purification device described in the utility model carries out smart purification, institute again to target protein after slightly purifying
The target protein purity of purification is high.
A kind of protein purification device described in the utility model is totally-enclosedization pipeline purifying plant, avoids protein empty
Gas pollutes, and keeps the activity of protein.
Embodiment 2
As shown in Fig. 2, the present embodiment difference from example 1 is that：
The compensator 103, chromatographic column 1, waste liquid tank 101 are sequentially connected, and 101 bottom of the waste liquid tank passes through circulation conduit
105 are connected with 103 top of the compensator, and a circulating pump 104, the compensator 103 and layer are arranged on the circulation conduit 105
Analysis column 1 is respectively provided with solenoid valve 7 between chromatographic column 1 and waste liquid tank 101, waste liquid tank 101 and circulating pump 104, the circulating pump 104,
Solenoid valve 7 is electrically connected with control system.The solenoid valve 7, circulating pump 104, the compensator are opened by the control system
Equilibrium liquid in 103 is flowed into from 1 top of the chromatographic column, and bottom outflow, into the waste liquid tank 101, the circulating pump 104 will
Waste liquid in the waste liquid tank 101 is pumped into the compensator 103, and realization puts down gel layer in the chromatographic column 1 using waste liquid
Weighing apparatus.
The chromatographic column 1, pH regulating reservoirs 2 are connected with protein concentration measuring appliance respectively, the protein concentration measuring appliance
It is electrically connected with the control system.The control system includes controlling the display panel, described to control the display panel the control electromagnetism
The relevant parameter of the solenoid valve 7, peristaltic pump 6 is arranged in the opening and closing of valve 7, peristaltic pump 6, shows the pH, protein compression angle value.
Protein example to be purified in the sample addition zone 106 enters in the chromatographic column 1, and the protein concentration measures
6 three of peristaltic pump between device, control system, chromatographic column 1, pH regulating reservoirs 2 cooperates, will be in chromatographic column 1 with suitable pump speed
Protein after screening is pumped into the pH regulating reservoirs 2, the control system, protein concentration measuring appliance, pH regulating reservoirs 2, thoroughly
Analyse column 3 between 6 three of peristaltic pump cooperate, with suitable pump speed by pH regulating reservoirs 2 after the adjustment after target protein pump
Enter in the dialysis cartridge 3.
A kind of protein purification device described in the utility model can recycle equilibrium liquid, improve the utilization of equilibrium liquid
Rate reduces the pollution to environment, cost-effective.
A kind of protein purification apparatus structure described in the utility model is simple and convenient to operate, can automate purification target
Protein, work efficiency is high.
While there has been shown and described that the embodiments of the present invention, for the ordinary skill in the art,
It is appreciated that can these embodiments be carried out with a variety of variations in the case where not departing from the principles of the present invention and spirit, repaiied
Change, replace and modification, the scope of the utility model are defined by the appended claims and the equivalents thereof.
Claims (10)
1. a kind of protein purification device, including sequentially connected chromatographic column, pH regulating devices, it is characterised in that：The pH tune
Regulating device is sequentially connected dialysis cartridge, purifying plant, collecting tank, and the purifying plant includes purifying slot, the two of the purifying slot
Side is respectively set passes through conducting wire, switch and control system with the positive and negative pole plate of the metal of external insulation, the positive and negative pole plate of metal
In high voltage power supply connect to generate electric field.
2. a kind of protein purification device according to claim 1, it is characterised in that：The top of the chromatographic column and sample-adding
Room is connected.
3. a kind of protein purification device according to claim 1, it is characterised in that：The chromatography top end also with balance
Tank is connected, and the bottom of the chromatographic column is connected with waste liquid tank.
4. a kind of protein purification device according to claim 3, it is characterised in that：The waste liquid tank passes through circulation conduit
It is connected with the compensator, circulating pump is set on the circulation conduit, the circulating pump is electrically connected with the control system.
5. a kind of protein purification device according to claim 1, it is characterised in that：The pH regulating devices include liquid feeding
Pond, pH regulating reservoirs, the pH measuring systems being connected with pH regulating reservoirs, the pH regulating reservoirs pass through a solenoid valve and the liquid-adding cell bottom
Portion is connected, and the pH measuring systems, solenoid valve and the control system are electrically connected.
6. a kind of protein purification device according to claim 1, it is characterised in that：Setting is semi-transparent in the dialysis cartridge
The dialysis cartridge is divided into solution storage chamber, protein storage room, the solution storage chamber and holding vessel phase by film, the semi-permeable membrane
Even, the protein storage room is connected with the top of the purifying plant, the bottom of the purifying plant and the collecting tank phase
Even.
7. a kind of protein purification device according to claim 1, it is characterised in that：The chromatographic column is adjusted with pH and is filled
It sets, be respectively provided with peristaltic pump between pH regulating devices and dialysis cartridge, dialysis cartridge and purifying plant, the peristaltic pump is with the control
System is electrically connected.
8. a kind of protein purification device according to claim 1, it is characterised in that：The side wall setting one of the chromatographic column
A or multiple liquid outlets, the liquid outlet are sequentially connected the pH regulating devices, dialysis cartridge, purifying plant, collecting tank.
9. a kind of protein purification device according to claim 1, it is characterised in that：The dialysis cartridge also with protein compression
It spends measuring appliance to be connected, the protein concentration measuring appliance is electrically connected with control system.
10. a kind of protein purification device according to claim 1, it is characterised in that：The pH regulating devices further include
The protein concentration measuring appliance being connected with the pH regulating reservoirs, the protein concentration measuring appliance are electrically connected with control system.
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|---|---|---|---|
| CN201721861786.1U CN207958195U (en) | 2017-12-27 | 2017-12-27 | A kind of protein purification device |
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| CN201721861786.1U CN207958195U (en) | 2017-12-27 | 2017-12-27 | A kind of protein purification device |
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| CN207958195U true CN207958195U (en) | 2018-10-12 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111574584A (en) * | 2020-05-21 | 2020-08-25 | 中国科学院生物物理研究所 | Full-automatic protein purification system device and application thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111574584A (en) * | 2020-05-21 | 2020-08-25 | 中国科学院生物物理研究所 | Full-automatic protein purification system device and application thereof |
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Effective date of registration: 20190711 Address after: 518000 Xufei Garden Building B 2428, 25 Bagua Second Road, Futian District, Shenzhen City, Guangdong Province Patentee after: Hong Bin Address before: 330098 No. 115 Ziyang Avenue (Administrative Building) 411-28, Nanchang High-tech Industrial Development Zone, Nanchang City, Jiangxi Province Patentee before: Nanchang Pu Wei Biotechnology Co., Ltd. |
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