A kind of protein purification deviceTechnical 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 proteinApparatus has a major impact, and is present in due to protein in complicated biosystem, and itself is unstable, heat or the certain solvents of chanceIt 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 notWith in density gradient layer, achieving the purpose that separation and protein purification, therefore ultracentrifugation or gradient density centrifugal isolate and purifyMethod 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 eggIt 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 changeMatter antigenic component precipitates, to achieve the purpose that purifying, most common method are salt precipitation methods.The principle of salting out method is albumenThe solubility of matter in aqueous solution depends on the hydrone number around protein molecule surface ion, that is, mainly by albumenWhat the case where matter molecule periphery hydrophilic radical and the degree and protein molecule of water formation hydration shell carry charge determined.AlbumenAfter neutral salt is added in matter solution, since the affinity of neutral salt and hydrone is more than protein, cause around protein moleculeHydrated sheath weaken so that disappear.Meanwhile neutral salt is added after protein solution since ionic strength changes, protein tableThe 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 eachThe protein of different solubility of the kind protein in different salinity, the salting liquid precipitation of different saturation is different, to makeCome 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 notHeight 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 knotThe substance of structure is packed into chromatographic column after solution equilibria appropriate.A kind of sample solution containing various molecules slowly flows throughWhen chromatographic column, macromolecular substances are not easily accessible the micropore of gel particle, can only be distributed between particle, therefore downward in elutionMobile speed, is eluted at first.Small-molecule substance can also pass into solidifying other than it can be spread in gel particle gapIn the micropore of glue particle, the speed that when elution moves down is slower, is then eluted.Therefore, protein molecule presses molecular sizeIt 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 andThe 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 adsorbedOn 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 SPARaw specific binding, remaining ingredient cannot be in combination.After chromatographic column is fully eluted, change the ionic strength or pH of eluentValue, 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 proteinThe 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, describedProtein 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 successivelyDialysis cartridge, purifying plant, collecting tank are connected, the purifying plant includes purifying slot, is respectively set in the both sides of the purifying slotWith 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 systemVoltage 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 purifiedProtein example carries out chromatography screening into the chromatographic column, and the protein after screening enters the pH regulating reservoirs, the pH tuneTarget 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 electricityIt is not acted on by electric field force in, therefore, target protein falls along straight line under gravity, into collecting tank, and itsRemaining 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 outLiquid 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, instituteThe 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 compensatorIt 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 systemCirculating pump is stated, the equilibrium liquid in the compensator flows at the top of the chromatographic column, bottom outflow, into the waste liquid tank, instituteIt 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 columnWeighing 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 tuneThe 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 measuredSystem is electrically connected with the control system.Protein after screening enters the pH regulating reservoirs, and the pH-meter measures the pHPH in regulating reservoir, the control system pass through the electromagnetism valve regulation institute between the pH regulating reservoirs, liquid-adding cell according to measurement dataThe 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 proteinIt 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 dialysisColumn is divided into solution storage chamber, protein storage room, and the solution storage chamber is connected with holding vessel, the protein storage room and instituteThe 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 withIt 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 columnMeasuring appliance is spent, the protein concentration measuring appliance is electrically connected with control system, and the protein concentration measuring appliance measures the chromatographyThe concentration of target protein and the control system is transferred data in column, when the concentration of target protein reaches setting valueAfterwards, the wriggling pump work between the chromatographic column, regulating device, the control system are arranged automatically according to the concentration of target proteinThe 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 reservoirsMeasurement of concetration device, the protein concentration measuring appliance are electrically connected with the control system, and the control system is according in pH regulating reservoirsProtein 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 instituteThe 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 throughOne 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 panelThe 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, albumenThe 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, purificationDevice, 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 liquidRate 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 quiltAir 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 ofPurify 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 loadedRoom；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 retouchedIt 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 thisEmbodiment in utility model, those of ordinary skill in the art obtained under the conditions of not making creative work it is all itsIts 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 cell201 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 dialysisColumn 3 divides for solution storage chamber, protein storage room, and the solution storage chamber and 303 top of the holding vessel are connected, the albumenMatter 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.InstituteIt 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 4State 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 dialysisIt 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 slotBoth 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 metalHigh voltage power supply in system connects to generate electric field.The pH measuring systems, solenoid valve 7, peristaltic pump 6, switch and control systemIt is electrically connected.
Protein example to be purified is put into sample addition zone 106, the peristaltic pump 6, electromagnetism are opened by control systemValve 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 chromatographyDrop 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 adjustedProtein 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 27 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 2Target 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 balanceProtein 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 remainedRemaining 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 instituteIt 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 eggWhite matter is not acted in the electric field by electric field force, and therefore, target protein falls along straight line under gravity, into instituteCollecting tank 5 is stated, and remaining protein is deflected for charged protein by electric field force effect, on positive and negative sheet metal pole plateEnrichment.
A kind of protein purification device described in the utility model carries out smart purification, institute again to target protein after slightly purifyingThe 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 emptyGas 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 conduit105 are connected with 103 top of the compensator, and a circulating pump 104, the compensator 103 and layer are arranged on the circulation conduit 105Analysis 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 systemEquilibrium 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 willWaste 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 liquidWeighing apparatus.
The chromatographic column 1, pH regulating reservoirs 2 are connected with protein concentration measuring appliance respectively, the protein concentration measuring applianceIt is electrically connected with the control system.The control system includes controlling the display panel, described to control the display panel the control electromagnetismThe 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 measures6 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 speedProtein after screening is pumped into the pH regulating reservoirs 2, the control system, protein concentration measuring appliance, pH regulating reservoirs 2, thoroughlyAnalyse column 3 between 6 three of peristaltic pump cooperate, with suitable pump speed by pH regulating reservoirs 2 after the adjustment after target protein pumpEnter 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 liquidRate 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 targetProtein, 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, repaiiedChange, replace and modification, the scope of the utility model are defined by the appended claims and the equivalents thereof.