CN223586921U - A protein free-flow isoelectric focusing electrophoresis separation device - Google Patents
A protein free-flow isoelectric focusing electrophoresis separation deviceInfo
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- CN223586921U CN223586921U CN202422935397.5U CN202422935397U CN223586921U CN 223586921 U CN223586921 U CN 223586921U CN 202422935397 U CN202422935397 U CN 202422935397U CN 223586921 U CN223586921 U CN 223586921U
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Abstract
本实用新型公开了一种蛋白质自由流等电聚焦电泳分离装置，包括冷液板，冷液板顶部安装有硅胶导热垫，硅胶导热垫顶部设置有电泳分离室，电泳分离室包括分离室下板，分离室下板与硅胶导热垫连接，分离室下板上方设置有分离室上板，分离室上板顶部连接有一对对称的电极室，电极室内贯穿连接有碳棒，电极室底部与分离室上板顶部皆开设有相同大小的矩形孔，分离室上板底部位于矩形孔对应处分别粘接有用以隔绝电极液与运行缓冲溶液的阴离子交换膜和阳离子交换膜，分离室上板和分离室下板之间设置有硅胶垫片，分离室上板和分离室下板之间通过夹子连接，分离室上板短边两侧皆分别开设有若干个分离室进液口和分离室出液口。
This utility model discloses a protein free-flow isoelectric focusing electrophoresis separation device, including a cold liquid plate, a silicone thermal pad installed on the top of the cold liquid plate, an electrophoresis separation chamber on the top of the silicone thermal pad, the electrophoresis separation chamber including a lower separation chamber plate connected to the silicone thermal pad, an upper separation chamber plate above the lower separation chamber plate, a pair of symmetrical electrode chambers connected to the top of the upper separation chamber plate, carbon rods penetrating through the electrode chambers, rectangular holes of the same size opened at the bottom of the electrode chambers and the top of the upper separation chamber plate, anion exchange membrane and cation exchange membrane for isolating the electrode liquid from the running buffer solution respectively bonded at the corresponding positions of the rectangular holes at the bottom of the upper separation chamber plate, a silicone gasket is placed between the upper separation chamber plate and the lower separation chamber plate, the upper separation chamber plate and the lower separation chamber plate are connected by a clip, and several separation chamber inlets and separation chamber outlets are opened on both sides of the short side of the upper separation chamber plate.
Description
Technical Field
The utility model relates to the technical field of protein separation, in particular to a protein free flow isoelectric focusing electrophoresis separation device.
Background
With the continuous development of biotechnology and life science research, protein separation and purification technology has become an important topic in life science research. Isoelectric focusing (Isoelectric Focusing, IEF) is used as a high-efficiency separation method, and can be widely applied to the fields of proteomics, clinical diagnosis, drug development and the like because of high-resolution separation based on isoelectric points (pI) of proteins. Conventional isoelectric focusing techniques generally employ a fixed medium (e.g., gel or membrane) to maintain the gradient of the electric field, and electrophoretically focus the protein in a pH gradient according to its isoelectric point. However, the use of a fixed medium limits the flexibility and accuracy of the separation process and is inefficient in high throughput analysis. In recent years, the protein Free flow isoelectric focusing technique (Free-Flow Isoelectric Focusing, FFIEF) has been developed as a novel separation method. This method does not rely on a solid medium, but rather by the action of an electric field in an electrophoretic separation chamber, causes the sample to migrate and focus along a pH gradient in a flowing liquid medium. This technique has many advantages over conventional methods, such as higher resolution, less sample loss, better reproducibility, and greater sample throughput. In particular, protein free-flow isoelectric focusing techniques show unique advantages when processing complex samples and performing large scale separations.
However, the existing protein free flow isoelectric focusing electrophoresis device generally has the problems of inaccurate temperature control, low separation efficiency, complex equipment structure and the like. The problems seriously affect the stability and the repeatability of the separation result, and in practical application, the operation difficulty is high, the problems of uneven electric field, protein denaturation and the like are easy to occur, and the separation effect is further reduced.
Disclosure of utility model
Aiming at the problems, the utility model provides a protein free-flow isoelectric focusing electrophoresis separation device, which effectively solves the problems of inaccurate temperature control, low separation efficiency and complex equipment structure of the existing protein free-flow isoelectric focusing electrophoresis device.
The utility model adopts the following technical scheme that the protein free flow isoelectric focusing electrophoresis separation device comprises a cold liquid plate, wherein a silica gel heat conduction pad is arranged at the top of the cold liquid plate, an electrophoresis separation chamber is arranged at the top of the silica gel heat conduction pad, the electrophoresis separation chamber comprises a separation chamber lower plate, the separation chamber lower plate is connected with the silica gel heat conduction pad, a separation chamber upper plate is arranged above the separation chamber lower plate, the top of the separation chamber upper plate is connected with a pair of symmetrical electrode chambers, carbon rods are connected in the electrode chambers in a penetrating manner, rectangular holes with the same size are formed in the bottoms of the electrode chambers and the tops of the separation chamber upper plate, an anion exchange membrane and a cation exchange membrane for isolating electrode liquid from running buffer solution are respectively bonded at the bottoms of the rectangular holes, a silica gel gasket is arranged between the separation chamber upper plate and the separation chamber lower plate, the separation chamber upper plate is connected with the separation chamber lower plate through clamps, and a plurality of separation chamber liquid inlets and separation chamber liquid outlets are respectively formed in two sides of the short sides of the separation chamber upper plate.
Further, the cold liquid plate is externally connected with a low-temperature cooling circulating pump.
Furthermore, round holes for electrode liquid circulation are formed in two sides of the top of the electrode chamber, an electrode liquid container and an electrode liquid driving pump are connected between the two round holes through pipelines, one end of a carbon rod above the anion exchange membrane is connected with the negative electrode of an external electrophoresis apparatus power supply, and one end of the carbon rod above the cation exchange membrane is connected with the positive electrode of the external electrophoresis apparatus power supply.
Further, the liquid inlet of the separation chamber is connected with a gas-liquid buffer chamber through a pipeline, the gas-liquid buffer chamber is filled with background buffer solution, and the liquid outlet of the separation chamber is connected with a collecting device through a pipeline.
Further, the inside of the electrode liquid container which is in circulation exchange with the electrode chamber above the anion exchange membrane is an alkaline solution, and the inside of the electrode liquid container which is in circulation exchange with the electrode chamber above the cation exchange membrane is an acidic solution.
Further, the thickness of the silica gel gasket, the thickness of the anion exchange membrane and the thickness of the cation exchange membrane are 1mm.
Furthermore, the lower plate of the separation chamber is made of glass, and the upper plate of the separation chamber is made of acrylic.
Further, one of the separation chamber inlets is connected with a protein sample to be separated through a sample driving pump.
The utility model has the advantage that by precisely controlling the electric field distribution in the electrophoretic separation chamber, the protein can be precisely focused according to its isoelectric point (pI). The acid-base property of the electrode liquid (through the circulation of the acidic and alkaline solutions) effectively controls the gradient of the electric field, and ensures the efficient separation and focusing of the protein. Through cold liquid board and cryocooling circulating pump (5), can maintain the required low temperature environment in the electrophoresis separation process effectively, prevent protein sample (11) denaturation or degradation, ensure the stability and the accuracy of experimental result. The anion exchange membrane and the cation exchange membrane are adopted to isolate the electrode liquid and the buffer liquid, so that the electrode liquid is prevented from polluting the buffer liquid, and the stability and the accuracy of the electrophoresis process are ensured. The membrane design ensures mutual isolation of the electrode liquid and the separation solution, and meanwhile, the uniformity of an electric field is not affected. The device is suitable for separating various proteins, and has excellent separation effect especially for proteins with different isoelectric points. By adjusting parameters such as an electric field, a buffer solution, a pH value and the like, complex protein mixtures can be accurately separated, and the method is widely applicable to the fields of biological research, proteomics analysis and the like.
Drawings
FIG. 1 is a schematic diagram of the structure of the present utility model;
FIG. 2 is a schematic view of the structure of the electrophoresis separation chamber of the present utility model;
fig. 3 is a schematic structural view of the present utility model.
In the figure, a 1-cold liquid plate, a 2-silica gel heat conduction pad, a 3-electrophoresis separation chamber, a 31-separation chamber lower plate, a 32-separation chamber upper plate, a 33-electrode chamber, a 34-carbon rod, a 35-anion exchange membrane, a 36-cation exchange membrane, a 37-silica gel pad, a 38-separation chamber liquid inlet, a 39-separation chamber liquid outlet, a 4-round hole, a 5-cryocooling circulating pump, a 6-electrode liquid container, a 7-electrode liquid driving pump, an 8-external electrophoresis apparatus power supply, a 9-gas-liquid buffer chamber, a 10-sample driving pump and an 11-protein sample are arranged.
Detailed Description
In order that those skilled in the art will better understand the present utility model, a technical solution in the embodiments of the present utility model will be clearly and completely described below with reference to the accompanying drawings in which it is apparent that the described embodiments are only some embodiments of the present utility model, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the present utility model without making any inventive effort, shall fall within the scope of the present utility model.
In the description of the present utility model, it should be understood that the terms "longitudinal", "transverse", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc. indicate orientations or positional relationships based on the drawings, are merely for convenience in describing the present utility model and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present utility model.
Referring to fig. 1-3, an isoelectric focusing electrophoresis separation device for protein free flow comprises a cold liquid plate 1, a silica gel heat conduction pad 2 is installed at the top of the cold liquid plate 1, an electrophoresis separation chamber 3 is arranged at the top of the silica gel heat conduction pad 2, the electrophoresis separation chamber 3 comprises a separation chamber lower plate 31, the separation chamber lower plate 31 is connected with the silica gel heat conduction pad 2, a separation chamber upper plate 32 is arranged above the separation chamber lower plate 31, a pair of symmetrical electrode chambers 33 are connected at the top of the separation chamber upper plate 32, carbon rods 34 are connected in the electrode chambers 33 in a penetrating manner, rectangular holes with the same size are formed at the bottom of the electrode chambers 33 and the top of the separation chamber upper plate 32, an anion exchange membrane 35 and a cation exchange membrane 36 for isolating electrode liquid from running buffer solution are respectively adhered at the bottom of the separation chamber upper plate 32, gaskets 37 are arranged between the separation chamber upper plate 32 and the separation chamber lower plate 31, a plurality of liquid inlets 39 are formed at two sides of the separation chamber upper plate 32 through the connection of clamps.
The cold liquid plate 1 is externally connected with a low-temperature cooling circulating pump 5, so that the temperature of the cold liquid plate is effectively maintained, the temperature in the whole separation process is ensured to be in a proper range all the time, the protein separation process is optimized, and the degradation of samples caused by heat accumulation is avoided.
The round holes 4 for circulating the electrode liquid are formed in the two sides of the top of the electrode chamber 33, the electrode liquid container 6 and the electrode liquid driving pump 7 are connected between the two round holes 4 through pipelines, one end of the carbon rod 34 above the anion exchange membrane 35 is connected with the negative electrode of the external electrophoresis apparatus power supply 8, one end of the carbon rod 34 above the cation exchange membrane 36 is connected with the positive electrode of the external electrophoresis apparatus power supply 8, so that the electrode liquid can circulate in the electrode chamber, the concentration and pH value change of the electrode liquid are avoided, the electric field stability is facilitated, the efficiency and the accuracy of electrophoresis separation are improved, the electric field intensity in the electrophoresis process and the acid-base property of the electrode liquid can be accurately controlled, and the efficient separation under different experimental conditions is ensured.
The liquid inlet 38 of the separation chamber is connected with the gas-liquid buffer chamber 9 through a pipeline, the gas-liquid buffer chamber 9 is filled with background buffer solution, the liquid outlet 39 of the separation chamber is connected with the collecting device through a pipeline, and the background buffer solution can stably enter the separation chamber 3 to keep the stability of the electrophoresis buffer solution. The separation chamber outlet 39 is connected to a collection device to ensure that the separated sample can be efficiently collected and processed.
The alkaline solution is in the electrode liquid container 6 which is circularly exchanged with the electrode chamber 33 above the anion exchange membrane 35, the acidic solution is in the electrode liquid container 6 which is circularly exchanged with the electrode chamber 33 above the cation exchange membrane 36, and the alkaline and acidic electrode liquid containers are respectively configured according to the positions of the anions and the cation exchange membranes, so that the pH value of the electrode liquid can be effectively controlled in the separation process, the influence of pH fluctuation on the electrophoresis result is avoided, and the electric field strength and stability are ensured through independent circular exchange of the acidic and alkaline electrode liquid, thereby improving the separation effect of proteins, and especially when proteins with different pI values are processed, the separation resolution is higher.
The thickness of silica gel gasket 37 with anion exchange membrane 35 and cation exchange membrane 36 is 1mm, helps promoting the sealing performance of device, prevents that liquid from leaking, ensures that unexpected does not take place in the operation process, promotes the security of experiment.
The lower plate 31 of the separation chamber is made of glass, and the upper plate 32 of the separation chamber is made of acrylic, so that transparent observation of the electrophoresis process is ensured, and meanwhile, the durability is enough.
One of the separation chamber inlets 38 is connected with the protein sample 11 to be separated through the sample driving pump 10, so that the addition of the sample can be precisely controlled, and the separation efficiency of the sample and the controllability of the experiment are improved.
The working principle is that a background buffer solution fills a gas-liquid buffer chamber 9 and enters an electrophoresis separation chamber 3 through a separation chamber liquid inlet 38, the background buffer solution is phosphate buffer solution, the background solution provides proper ionic strength and pH environment in an electric field, the migration and focusing of proteins are facilitated, a cryocooling circulation pump 5 is started, the circulation of the cooling liquid is ensured, the set cryotemperature range is reached, a sample driving pump 10 is started, a protein sample 11 to be separated is added into the electrophoresis separation chamber 3 through the separation chamber liquid inlet 38, a proper amount of acidic solution or alkaline solution is added into two electrode liquid containers 6, the pH value of liquid at two sides of the electrode chamber meets the experiment requirement, an electrode liquid driving pump 7 is started, the circulating flow of the electrode liquid in the electrode chamber is ensured, the formation of an electric field is stabilized, the cathode of an external electrophoresis apparatus is connected to a carbon rod 34 on an anion exchange membrane 35, the anode is connected to the carbon rod 34 on the cation exchange membrane 36, proper voltage and current are set according to the experiment requirement, an external electrophoresis apparatus power supply 8 is started, the electric field is uniformly distributed in the electrophoresis separation chamber 3, when the electric field acts on the separation chamber 3, the protein and the isoelectric point of the protein and the isoelectric point of the electrophoresis apparatus are continuously moved to different pH values in the different directions along with the positive and negative electrode gradient and the positive and negative electrode gradient of the protein migration and the different pH values, the protein migration and the isoelectric point of the protein migration and the isoelectric point are continuously in the different pH values in the different potential gradient directions, respectively, the final electrophoresis separation region is realized, and are arranged along the electrophoresis direction in the separation chamber. The anion exchange membrane 35 and the cation exchange membrane 36 serve as key components in the electrophoresis separation system, play a role in isolating electrode liquid and buffer liquid, prevent the electrode liquid from polluting the electrophoresis buffer liquid, ensure the uniformity of an electric field and stabilize the electrophoresis process. The silica gel gasket 37 then acts as a seal and barrier between the upper and lower plates of the separation chamber, preventing leakage of solution, while helping to maintain overall system stability. Because protein separation is very sensitive to temperature, a cooling system (a cold liquid plate and a circulating pump) in the device is always kept at low temperature, so that protein is ensured not to be denatured or degraded in the separation process. After the electrophoretic separation is completed, the outlet 39 of the separation chamber is connected to a collection device, which allows the collection of the protein fraction focused at different isoelectric points.
It will be evident to those skilled in the art that the utility model is not limited to the details of the foregoing illustrative embodiments, and that the present utility model may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, the scope of the utility model being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Furthermore, it should be understood that although the present disclosure describes embodiments, not every embodiment is provided with a separate embodiment, and that this description is provided for clarity only, and that the disclosure is not limited to the embodiments described in detail below, and that the embodiments described in the examples may be combined as appropriate to form other embodiments that will be apparent to those skilled in the art.
Claims (8)
1. The protein free flow isoelectric focusing electrophoresis separation device comprises a cold liquid plate (1), and is characterized in that a silica gel heat conduction pad (2) is arranged at the top of the cold liquid plate (1), an electrophoresis separation chamber (3) is arranged at the top of the silica gel heat conduction pad (2), the electrophoresis separation chamber (3) comprises a separation chamber lower plate (31), the separation chamber lower plate (31) is connected with the silica gel heat conduction pad (2), a separation chamber upper plate (32) is arranged above the separation chamber lower plate (31), a pair of symmetrical electrode chambers (33) are connected at the top of the separation chamber upper plate (32), a carbon rod (34) is connected in the electrode chambers (33) in a penetrating manner, rectangular holes with the same size are formed at the bottom of the electrode chambers (33) and the top of the separation chamber upper plate (32), an anion exchange membrane (35) and a cation exchange membrane (36) which are used for isolating electrode liquid and running buffer solution are respectively bonded at the bottom of the separation chamber upper plate (32) corresponding to the rectangular holes, a separation chamber upper plate (32) and a separation chamber lower plate (31) are connected with a separation chamber (37) through a clamp (37), a plurality of separating chamber liquid inlets (38) and separating chamber liquid outlets (39) are respectively formed on two sides of the short side of the separating chamber upper plate (32).
2. The protein free flow isoelectric focusing electrophoresis separation device according to claim 1, wherein the cold liquid plate (1) is externally connected with a low-temperature cooling circulating pump (5).
3. The protein free flow isoelectric focusing electrophoresis separation device according to claim 2, wherein round holes (4) for circulating electrode liquid are formed in two sides of the top of the electrode chamber (33), an electrode liquid container (6) and an electrode liquid driving pump (7) are connected between the two round holes (4) through pipelines, one end of a carbon rod (34) above an anion exchange membrane (35) is connected with the negative electrode of an external electrophoresis apparatus power supply (8), and one end of the carbon rod (34) above a cation exchange membrane (36) is connected with the positive electrode of the external electrophoresis apparatus power supply (8).
4. The protein free flow isoelectric focusing electrophoresis separation device according to claim 3, wherein the liquid inlet (38) of the separation chamber is connected with a gas-liquid buffer chamber (9) through a pipeline, the gas-liquid buffer chamber (9) is filled with a background buffer solution, and the liquid outlet (39) of the separation chamber is connected with a collecting device through a pipeline.
5. The apparatus of claim 4, wherein an alkaline solution is contained in the electrode solution container (6) for circulating the electrode chamber (33) above the anion exchange membrane (35), and an acidic solution is contained in the electrode solution container (6) for circulating the electrode chamber (33) above the cation exchange membrane (36).
6. A protein free flow isoelectric focusing electrophoresis separation device according to claim 5, wherein the thickness of each of the silica gel pad (37) and the anion exchange membrane (35) and the cation exchange membrane (36) is 1mm.
7. The device of claim 6, wherein the lower plate (31) is made of glass and the upper plate (32) is made of acrylic.
8. A protein free-flow isoelectric focusing electrophoresis separation device according to claim 7 wherein one of said separation chamber fluid inlets (38) is connected to a protein sample (11) to be separated by a sample driven pump (10).
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| CN202422935397.5U CN223586921U (en) | 2024-11-29 | 2024-11-29 | A protein free-flow isoelectric focusing electrophoresis separation device |
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| CN202422935397.5U CN223586921U (en) | 2024-11-29 | 2024-11-29 | A protein free-flow isoelectric focusing electrophoresis separation device |
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