Enzyme separation device

Abstract

The utility model discloses an enzyme separation device, which relates to the field of separation and purification equipment and comprises a rack, a plurality of groups of temperature control clamping pieces are movably connected onto the rack, and an air pressure control column is clamped at the top of the rack through a first group of temperature control clamping pieces. A defoaming chromatographic column is clamped in the middle of the rack through a second group of temperature control clamping pieces; an inner cavity of the pneumatic control column is communicated with an inner cavity of the defoaming chromatographic column through a hose; and a collecting barrel is placed at the bottom of the rack and is positioned under the defoaming chromatographic column. The chromatographic device is simple in structure and reasonable in layout, and effectively solves the problems that a traditional chromatographic device does not have a temperature control function and the chromatographic effect is not ideal due to the fact that bubbles are generated.

Description

Technical Field

[0001] The utility model relates to the field of separation and purification equipment, in particular to an enzyme separation device. Background Art

[0002] In recent years, an effective method commonly used to separate and purify enzymes is affinity chromatography, which mainly uses the ability of enzymes to bind to substrates, inhibitors or coenzymes. This property can be used to separate and purify enzymes. First, a support is selected, such as agarose, to connect the substrate, competitive inhibitor or coenzyme to the support in the form of a covalent bond. Then, the solution containing the enzyme flows through a chromatography column containing a specific substrate, competitive inhibitor or coenzyme. The enzyme is retained on the support, and then the impurities that are not adsorbed are removed after sufficient washing. Then, a buffer containing a certain concentration of substrate, competitive inhibitor or coenzyme is used for competitive elution.

[0003] However, existing chromatography columns often do not have a temperature control function, which causes the enzyme to lose its activity due to inappropriate temperature during the separation and elution process; and, in the process of pouring the enzyme-containing solution or eluent into the chromatography column, bubbles will be generated in the internal matrix of the chromatography column, and the bubbles will cause the flow rate of the liquid to be eluted through the matrix to decrease, and the liquid will pass through the matrix unevenly, which will cause the target enzyme to be unable to fully bind to the matrix, affecting the enzyme separation and purification results.

[0004] Therefore, how to provide an enzyme separation device to achieve temperature control during enzyme affinity chromatography and avoid the generation of bubbles in the chromatography column, thereby improving the separation and purification effect of the enzyme, has become a difficult problem to be solved urgently by those skilled in the art. Summary of the invention

[0005] The utility model aims to provide an enzyme separation device to solve the problems that temperature control cannot be achieved during enzyme affinity chromatography and bubbles are easily generated in the internal matrix of the chromatography column.

[0006] In order to solve the above technical problems, the utility model adopts the following technical solutions:

[0007] The utility model discloses an enzyme separation device, comprising a frame, wherein a plurality of groups of temperature control clamps are movably connected to the frame, a gas pressure control column is clamped on the top of the frame by a first group of the temperature control clamps, a defoaming chromatography column is clamped in the middle of the frame by a second group of the temperature control clamps, and an inner cavity of the gas pressure control column and an inner cavity of the defoaming chromatography column are connected by a hose; a collection bucket is placed at the bottom of the frame, and the collection bucket is located directly below the defoaming chromatography column;

[0008] A PLC controller and an electric air pump are also installed at the bottom of the frame. The air outlet of the electric air pump is connected to the side wall of the air pressure control column through an air pipe, and the lumen of the air pipe is connected to the inner cavity of the air pressure control column; the PLC controller is electrically connected to the temperature control clamp, the air pressure control column, the defoaming chromatography column, and the electric air pump respectively.

[0009] Preferably, the frame includes a base, support rods are symmetrically installed on the left and right sides of the top of the base, and a top plate is inserted into the top of the support rods; two groups of temperature control clamps are installed on the support rods, and the PLC controller and the electric air pump are installed on the base.

[0010] Preferably, the temperature control clamp comprises a round tube and a round rod, the outer end of the round rod is slidably connected in the round tube, the inner end of the round rod is connected with an arc plate, the curvature of the arc plate matches the curvature of the side wall of the air pressure control column and the defoaming chromatography column respectively; a semiconductor refrigeration sheet is installed on the inner end surface of the arc plate, and the semiconductor refrigeration sheet is electrically connected to the PLC controller;

[0011] A sliding hole is provided on the side wall of the outer end of the circular tube, and the circular tube is sleeved on the support rod through the sliding hole. The outer end of the circular tube is connected with a first tightening screw through a thread, and the first tightening screw and the support rod are butted together; the side wall of the inner end of the circular tube is connected with a second tightening screw through a thread, and the second tightening screw and the circular rod are butted together.

[0012] Preferably, a first electric valve is installed at the input end of the air pressure control column, a second electric valve is installed at the output end of the air pressure control column, the output end of the air pressure control column is connected to the hose, and a barometer is installed on the side wall of the air pressure control column; the first electric valve, the second electric valve and the barometer are all electrically connected to the PLC controller.

[0013] Preferably, the debubble chromatography column comprises a column barrel and a barrel cover, the barrel cover is threadedly connected to the top of the column barrel, a plurality of limit sliding rods are evenly distributed on the inner wall of the column barrel in a circumferential manner, a debubble plate is slidably connected to the inside of the column barrel, and the debubble plate cooperates with the plurality of limit sliding rods and slides up and down along the limit sliding rods;

[0014] The input pipe at the top of the barrel cover is connected to the hose; an output pipe is arranged at the bottom center of the column barrel, a third electric valve is installed on the output pipe of the column barrel, and the third electric valve is electrically connected to the PLC controller.

[0015] Preferably, the main body of the debubble plate is a circular plate, and a plurality of slide grooves are evenly distributed in a circle on the side wall of the circular plate, and the slide grooves match the limiting slide rods; a plurality of sieve holes are opened on the body of the circular plate, and a plurality of gap rods are installed on the bottom surface of the circular plate, and the plurality of gap rods are evenly distributed in a circle, and a float is connected to the bottom of the plurality of gap rods.

[0016] Preferably, the circular plate is made of ultra-high molecular weight polyethylene material, and the float is made of EVA foam.

[0017] Compared with the prior art, the beneficial technical effects of the utility model are:

[0018] The utility model discloses an enzyme separation device, comprising a frame, wherein a plurality of groups of temperature control clamps are movably connected to the frame, a gas pressure control column is clamped on the top of the frame by a first group of the temperature control clamps, a defoaming chromatography column is clamped in the middle of the frame by a second group of the temperature control clamps, and an inner cavity of the gas pressure control column and an inner cavity of the defoaming chromatography column are communicated with each other by a hose; the utility model realizes temperature control of the gas pressure control column and the defoaming chromatography column by the temperature control clamps, thereby preventing the enzyme from losing activity due to inappropriate temperature during the separation process.

[0019] Before adding materials to the defoaming chromatography column, the utility model firstly passes the added liquid into the air pressure control column, and performs pressurization treatment on the added liquid to remove the internal bubbles of the added liquid.

[0020] The utility model discloses a debubble chromatography column, the interior of which is movably connected with a debubble plate, which has a buffering effect on the poured liquid, thereby preventing bubbles from being generated after the liquid is directly poured into the matrix. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] The utility model will be further described below in conjunction with the accompanying drawings.

[0022] Figure 1 This is a schematic diagram of the structure of the enzyme separation device of the utility model;

[0023] Figure 2 This is a cross-sectional view of the defoaming chromatography column of the utility model;

[0024] Figure 3 This is a schematic diagram of the structure of the defoaming plate of the utility model.

[0025] Description of reference numerals: 1. rack; 2. temperature control clamp; 3. air pressure control column; 4. defoaming chromatography column; 5. hose; 6. collection bucket; 7. PLC controller; 8. electric air pump; 9. air pipe;

[0026] 101. base; 102. support rod; 103. top plate;

[0027] 201, round tube; 202, round rod; 203, arc plate; 204, first tightening screw; 205, second tightening screw;

[0028] 301, first electric valve; 302, second electric valve; 303, barometer;

[0029] 401, column barrel; 402, barrel cover; 403, limit slide rod; 404, debubble plate; 405, third electric valve; 441, round plate; 442, slide groove; 443, sieve hole; 444, gap rod; 445, float. DETAILED DESCRIPTION

[0030] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0031] like Figure 1-3 As shown, an enzyme separation device comprises a frame 1, on which a plurality of groups of temperature control clamps 2 are movably connected, the top of the frame 1 is clamped with an air pressure control column 3 by a first group of the temperature control clamps 2, the middle of the frame 1 is clamped with a defoaming chromatography column 4 by a second group of the temperature control clamps 2, and the inner cavity of the air pressure control column 3 and the inner cavity of the defoaming chromatography column 4 are connected by a hose 5; a collecting bucket 6 is placed at the bottom of the frame 1, and the collecting bucket 6 is located directly below the defoaming chromatography column 4;

[0032] A PLC controller 7 and an electric air pump 8 are also installed at the bottom of the frame 1. The air outlet end of the electric air pump 8 is connected to the side wall of the air pressure control column 3 through an air pipe 9, and the lumen of the air pipe 9 is connected to the inner cavity of the air pressure control column 3; the PLC controller 7 is electrically connected to the temperature control clamp 2, the air pressure control column 3, the defoaming chromatography column 4, and the electric air pump 8 respectively.

[0033] Specifically, the frame 1 includes a base 101, and support rods 102 are symmetrically installed on the left and right sides of the top of the base 101, and a top plate 103 is inserted at the top of the support rod 102; two groups of temperature control clamps 2 are installed on the support rods 102, and the PLC controller 7 and the electric air pump 8 are installed on the base 101.

[0034] Specifically, the temperature control clamp 2 includes a round tube 201 and a round rod 202, the outer end of the round rod 202 is slidably connected to the round tube 201, and the inner end of the round rod 202 is connected to an arc plate 203, and the curvature of the arc plate 203 matches the curvature of the side wall of the air pressure control column 3 and the defoaming chromatography column 4 respectively; a semiconductor refrigeration sheet is installed on the inner end surface of the arc plate 203, and the semiconductor refrigeration sheet is electrically connected to the PLC controller 7;

[0035] A sliding hole is opened on the side wall of the outer end of the circular tube 201, and the circular tube 201 is sleeved on the support rod 102 through the sliding hole. The outer end of the circular tube 201 is connected with a first tightening screw 204 through a thread, and the first tightening screw 204 and the support rod 102 are abutted together; the side wall of the inner end of the circular tube 201 is connected with a second tightening screw 205 through a thread, and the second tightening screw 205 and the circular rod 202 are abutted together.

[0036] Specifically, the input end of the air pressure control column 3 is installed with a first electric valve 301, the output end of the air pressure control column 3 is installed with a second electric valve 302, the output end of the air pressure control column 3 is connected to the hose 5, and a barometer 303 is installed on the side wall of the air pressure control column 3; the first electric valve 301, the second electric valve 302 and the barometer 303 are all electrically connected to the PLC controller 7. Specifically, the input end of the air pressure control column 3 is connected with an external reagent bottle, and the external reagent bottle contains a crude enzyme solution, a substrate, a competitive inhibitor or a coenzyme. In addition, in a specific implementation, a funnel can also be inserted into the input end of the air pressure control column 3 to facilitate the liquid in the external reagent bottle to be poured into the column cavity of the air pressure control column 3 through the funnel.

[0037] Specifically, the debubble chromatography column 4 includes a column barrel 401 and a barrel cover 402, the barrel cover 402 is threadedly connected to the top of the column barrel 401, a plurality of limiting slide bars 403 are evenly distributed on the inner wall of the column barrel 401 in a circumferential manner, and a debubble plate 404 is slidably connected to the inside of the column barrel 401, and the debubble plate 404 cooperates with the plurality of limiting slide bars 403 and slides up and down along the limiting slide bars 403;

[0038] The input pipe at the top of the barrel cover 402 is connected to the hose 5; an output pipe is provided at the bottom center of the column barrel 401, and a third electric valve 405 is installed on the output pipe of the column barrel 401, and the third electric valve 405 is electrically connected to the PLC controller 7.

[0039] Specifically, the main body of the debubble plate 404 is a circular plate 441, and a plurality of slide grooves 442 are evenly distributed in a circle on the side wall of the circular plate 441, and the slide grooves 442 match the limiting slide rod 403; a plurality of sieve holes 443 are opened on the body of the circular plate 441, and a plurality of gap rods 444 are installed on the bottom surface of the circular plate 441, and the plurality of gap rods 444 are evenly distributed in a circle, and a float 445 is connected to the bottom of the plurality of gap rods 444.

[0040] Specifically, the circular plate 441 is made of ultra-high molecular weight polyethylene material, and the float 445 is made of EVA foam.

[0041] The use process of the utility model is as follows:

[0042] Clamping and installation operation: after the arc plate 203 in the temperature control clamp 2 respectively abuts against the air pressure control column 3 and the debubble chromatography column 4, the first tightening screw 204 and the second tightening screw 205 are tightened in sequence to complete the clamping operation of the air pressure control column 3 and the debubble chromatography column 4; thereafter, the output end of the air pressure control column 3 and the input end of the debubble chromatography column 4 are respectively connected through the hose 5 to complete the installation operation of the utility model.

[0043] Temperature control operation: The semiconductor refrigeration plate is controlled by a PLC controller to perform refrigeration operation. The refrigeration end of the semiconductor refrigeration plate abuts against the side walls of the air pressure control column 3 and the debubble chromatography column 4, and the temperature of the air pressure control column 3 and the debubble chromatography column 4 is controlled by heat conduction.

[0044] Pressurization operation: After the liquid is passed into the air pressure control column 3, the PLC controller controls the first electric valve 301 and the second electric valve 302 to close, and controls the electric air pump 8 to work according to the reading of the barometer 303 until the reading of the barometer 303 reaches a preset threshold, and uses high air pressure to remove bubbles in the added liquid.

[0045] Separation and purification operation: the liquid after the pressurized debubble treatment is passed into the debubble chromatography column 4, and the PLC controller reasonably controls the opening and closing of the first electric valve 301, the second electric valve 302 and the third electric valve 405, so that the required added solution is passed into the debubble chromatography column 4; after the liquid is passed into the debubble chromatography column 4, it is buffered by the debubble plate 404 and then flows into the original matrix in the debubble chromatography column 4, so as to avoid bubbles generated after the liquid is directly poured into the matrix.

[0046] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0047] The embodiments described above are only descriptions of the preferred methods of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should fall within the protection scope determined by the claims of the present invention.

Claims

1. An enzyme separation device, characterized in that: The invention comprises a frame (1), wherein a plurality of groups of temperature control clamps (2) are movably connected to the frame (1), a gas pressure control column (3) is clamped at the top of the frame (1) by a first group of the temperature control clamps (2), a defoaming chromatography column (4) is clamped at the middle of the frame (1) by a second group of the temperature control clamps (2), and the inner cavity of the gas pressure control column (3) and the inner cavity of the defoaming chromatography column (4) are connected by a hose (5); a collecting bucket (6) is placed at the bottom of the frame (1), and the collecting bucket (6) is located directly below the defoaming chromatography column (4); A PLC controller (7) and an electric air pump (8) are also installed at the bottom of the frame (1); the air outlet of the electric air pump (8) is connected to the side wall of the air pressure control column (3) through an air pipe (9), and the lumen of the air pipe (9) is connected to the inner lumen of the air pressure control column (3); the PLC controller (7) is electrically connected to the temperature control clamp (2), the air pressure control column (3), the defoaming chromatography column (4), and the electric air pump (8), respectively.

2. The enzyme separation device according to claim 1, characterized in that: The frame (1) comprises a base (101), support rods (102) are symmetrically mounted on the left and right sides of the top of the base (101), and a top plate (103) is plugged into the top of the support rod (102); two groups of temperature control clamps (2) are mounted on the support rods (102), and the PLC controller (7) and the electric air pump (8) are mounted on the base (101).

3. The enzyme separation device according to claim 2, characterized in that: The temperature control clamp (2) comprises a circular tube (201) and a round rod (202), the outer end of the round rod (202) is slidably connected in the circular tube (201), the inner end of the round rod (202) is connected to an arc plate (203), the curvature of the arc plate (203) matches the curvature of the side wall of the air pressure control column (3) and the defoaming chromatography column (4); a semiconductor refrigeration plate is installed on the inner end surface of the arc plate (203), and the semiconductor refrigeration plate is electrically connected to the PLC controller (7); A sliding hole is provided on the side wall of the outer end of the circular tube (201), and the circular tube (201) is sleeved on the support rod (102) through the sliding hole. The outer end of the circular tube (201) is connected to a first tightening screw (204) through a thread, and the first tightening screw (204) and the support rod (102) are in contact with each other; a second tightening screw (205) is connected to the side wall of the inner end of the circular tube (201) through a thread, and the second tightening screw (205) and the circular rod (202) are in contact with each other.

4. The enzyme separation device according to claim 3, characterized in that: The input end of the air pressure control column (3) is installed with a first electric valve (301), the output end of the air pressure control column (3) is installed with a second electric valve (302), the output end of the air pressure control column (3) is connected to the hose (5), and a barometer (303) is installed on the side wall of the air pressure control column (3); the first electric valve (301), the second electric valve (302) and the barometer (303) are all electrically connected to the PLC controller (7).

5. The enzyme separation device according to claim 4, characterized in that: The debubble chromatography column (4) comprises a column barrel (401) and a barrel cover (402), wherein the barrel cover (402) is threadedly connected to the top of the column barrel (401), a plurality of limiting slide bars (403) are evenly distributed on the inner wall of the column barrel (401) in a circular shape, a debubble plate (404) is slidably connected to the interior of the column barrel (401), and the debubble plate (404) cooperates with the plurality of limiting slide bars (403) and slides up and down along the limiting slide bars (403); The input pipe at the top of the barrel cover (402) and the hose (5) are connected together; an output pipe is provided at the bottom center of the column barrel (401), and a third electric valve (405) is installed on the output pipe of the column barrel (401), and the third electric valve (405) is electrically connected to the PLC controller (7).

6. The enzyme separation device according to claim 5, characterized in that: The main body of the debubble plate (404) is a circular plate (441), and a plurality of slide grooves (442) are evenly distributed in a circle on the side wall of the circular plate (441), and the slide grooves (442) match the limiting slide rod (403); a plurality of sieve holes (443) are opened on the plate body of the circular plate (441), and a plurality of gap rods (444) are installed on the bottom surface of the circular plate (441), and the plurality of gap rods (444) are evenly distributed in a circle, and a float (445) is connected to the bottom of the plurality of gap rods (444).

7. The enzyme separation device according to claim 6, characterized in that: The circular plate (441) is made of ultra-high molecular weight polyethylene material, and the floating body (445) is made of EVA foam.

Images