A protease chromatography extraction device

Abstract

The invention discloses a protease chromatography extraction device, relates to the technical field of protease extraction, and aims to solve the technical problem of low extraction efficiency of the protease chromatography extraction device, including a chromatography column and a rotating mechanism, the chromatography column is hollow inside to form an installation cavity, a column A is fixedly provided at the top of the installation cavity, three through holes A are provided in an annular equidistant structure on the column A, a column B is fixedly provided at the bottom of the installation cavity, three through holes B are provided in an annular equidistant structure on the column B, a gel mechanism is arranged in the gap between the column A and the column B, and the gel mechanism is rotatably connected to the installation cavity; the gel mechanism includes three gel cavities, and the two ends of the three gel cavities are respectively connected with the three through holes A and the three through holes B to respectively form a mixed liquid filtration channel, a protease elution channel and a cleaning channel; the rotating mechanism is arranged at the top of the chromatography column, and the output end of the rotating mechanism is connected to the gel mechanism. The invention realizes the automatic continuous extraction effect of protease, and has the advantage of high protease extraction efficiency.

Description

Technical Field

[0001] The present invention relates to the technical field of protease extraction, and more specifically to a protease chromatography extraction device. Background Art

[0002] Protease is a general term for a class of enzymes that hydrolyze protein peptide chains. According to the way they degrade polypeptides, they are divided into two categories: endopeptidases and terminal peptidases. The former can cut large molecular weight polypeptide chains from the middle to form smaller molecular weight proteins and peptidases; the latter can be divided into carboxypeptidases and aminopeptidases, which hydrolyze polypeptide chains one by one from the free carboxyl end or free amino end of the polypeptide to generate amino acids.

[0003] The existing protease production process includes raw material preparation, extraction, centrifugal separation, chromatographic separation, concentration and drying, wherein raw material preparation is to collect enzyme sources and use crushing equipment to crush them; extraction is to use stirring extraction equipment to add extracting agents for stirring extraction to obtain a protease mixture; centrifugal separation is to centrifuge and filter the protease supernatant to remove the precipitate and obtain the protease supernatant; chromatographic separation is to use chromatographic separation equipment to take the supernatant for chromatographic separation and collect the protease eluate; concentration and drying is to concentrate the eluate to obtain a protease solution, which is then dried to obtain a protease product.

[0004] The chromatographic extraction methods of proteases mainly include gel filtration (molecular sieve chromatography), ion exchange chromatography, affinity chromatography and countercurrent chromatography. The principle of gel filtration chromatography is to utilize the degree of adsorption of gel particles to different molecules to extract proteases. However, the existing protease chromatographic extraction equipment simply fills the gel particles in the chromatography column cavity. The extraction process is to fill the chromatography column cavity with gel particles of an appropriate height to form a gel layer according to factors such as the specific protease solution concentration, and the protease supernatant is removed from the gel chromatography column at the top of the chromatography column. The liquid and macromolecular substances fall through the gel layer, and the protease is adsorbed in the gel layer. The liquid and the protease in the gel layer are then taken away by the eluent, and the protease eluent is extracted for subsequent analysis and purification. Finally, the gel layer is rinsed with a cleaning solution to rinse off the residual substances in the gel layer. The above-mentioned protease chromatographic extraction equipment has a single function, a simple chromatography column structure, and a complicated chromatographic extraction process of the protease with low efficiency. In view of this, we propose a protease chromatographic extraction equipment. Summary of the invention

[0005] The object of the present invention is to provide a protease chromatography extraction device to solve the technical problem of low extraction efficiency of the protease chromatography extraction device.

[0006] In order to solve the above technical problems, the present invention provides the following technical solutions: a protease chromatography extraction device, comprising a chromatography column and a rotating mechanism, wherein the interior of the chromatography column is hollow to form an installation cavity, a column A is fixedly arranged on the top of the installation cavity, and three through holes A are provided on the column A in an annular structure with equal spacing, wherein the gap distance between any two through holes A is greater than the diameter of the through holes A, three flow guide tubes A are connected to the top of the chromatography column, and the three flow guide tubes A are respectively connected to the tops of the three through holes A, a column B is fixedly arranged on the bottom of the installation cavity, and three through holes B are provided on the column B in an annular structure with equal spacing, and the through holes B are adapted to the through holes A, and three flow guide tubes B are connected to the bottom of the chromatography column, and the three flow guide tubes B are respectively connected to the bottoms of the three through holes B, a gel mechanism is arranged in the gap between the column A and the column B, and the gel mechanism is rotatably connected to the installation cavity;

[0007] The gel mechanism comprises three gel chambers, and the two ends of the three gel chambers are respectively connected to the three through holes A and the three through holes B to form a mixed liquid filtration channel, a protease elution channel and a cleaning channel respectively;

[0008] The rotating mechanism is arranged at the top of the chromatography column, and the output end of the rotating mechanism passes through the chromatography column and the column A in sequence and is connected to the gel mechanism. The present invention designs the chromatography column, the rotating mechanism, the column A, the column B and the gel mechanism structure, so that the two ends of the three gel cavities are respectively connected with the three through holes A and the three through holes B to respectively form a mixed liquid filtration channel, a protease elution channel and a cleaning channel. When the protease is extracted, the protease mixed liquid is filtered from the mixed liquid filtration channel, and the protease is left on the gel particles. The gel mechanism is driven to rotate by the rotating mechanism, so that the gel cavity originally in the mixed liquid filtration channel rotates to the protease elution channel and forms a new channel, and the protease in the gel cavity is washed out by the eluent to obtain the protease eluent, so as to be analyzed and purified later, and the gel cavity originally in the protease elution channel is eluted and rotated to the cleaning channel and forms a new channel, and the cleaning channel is cleaned by the cleaning liquid, and the gel cavity originally in the cleaning channel that has been cleaned rotates to the mixed liquid filtration channel to form a new channel, so as to achieve the automatic continuous extraction effect of the protease, greatly save time, and solve the technical problem of low extraction efficiency of the protease chromatography extraction equipment.

[0009] Preferably, the rotating mechanism includes a motor and a connecting shaft, the motor is fixedly mounted at the top of the chromatography column, the connecting shaft is passed through the center of the column A, and the connecting shaft is rotationally connected to the column A, and the top end of the connecting shaft passes through the top of the chromatography column and is fixedly connected to the motor output shaft.

[0010] Preferably, three embedding grooves A are opened at the bottom end of the column A relative to the three through holes A, and a sealing ring A is embedded in the embedding groove A. The bottom end of the sealing ring A is in contact with the top end of the gel mechanism, and a snap-in groove A is opened at the center of the bottom end of the column A.

[0011] Preferably, three embedding grooves B are provided at the top of the column B relative to the three through holes B, and sealing rings B are embedded in the embedding grooves B, and the sealing rings B are in contact with and connected to the bottom end of the gel mechanism 5;

[0012] Wherein, the sealing ring A and the sealing ring B are both made of polytetrafluoroethylene.

[0013] Preferably, the gel mechanism comprises a gel bottom column and a gel top plate, the gel bottom column is rotatably connected to the mounting cavity through a plurality of rotating rings A, a rotating groove is provided at the center position of the top end of the gel bottom column, and a movable groove is provided at the bottom end of the rotating groove, a mounting hole is provided on the gel bottom column relative to the three through holes B, a gel assembly is arranged on the mounting hole, the three gel assemblies are linked by a linkage assembly, an adjustment assembly is arranged on the linkage assembly, the gel top plate is arranged above the gel bottom column, the gel top plate and the gel bottom column are fixedly connected by three connecting columns arranged in a ring-shaped equidistant structure, the top end of the gel top plate is in contact with the sealing ring A, three through holes C are provided on the gel top plate relative to the three through holes A, and a snap-in groove B is provided at the center position of the gel top plate.

[0014] Preferably, the gel assembly includes a mounting top ring and a mounting bottom ring, the mounting top ring is rotatably arranged at the top end of the mounting hole, the mounting top ring is rotatably connected to the gel top plate via a swivel B, the bottom end of the mounting top ring is an annular structure with equal spacing and is provided with a plurality of inclined guide grooves, the mounting bottom ring is fixedly arranged at the bottom end of the mounting hole, the bottom end of the mounting bottom ring is in contact with the sealing ring B, the mounting bottom ring is provided with a plurality of centripetal guide grooves in an annular structure with equal spacing relative to the number of the inclined guide grooves, a bearing mesh plate is fixedly installed on the top end of the ring cavity of the mounting bottom ring, and the centripetal guide groove is movably connected to the inclined guide groove via a blocking unit.

[0015] Preferably, the enclosure unit comprises a movable column A and a movable column B, wherein the movable column A is movably arranged on the oblique guide groove, and the movable column B is movably arranged on the centripetal guide groove, and the movable column A is rotatably connected to the movable column B via a rotating rod, and a connecting plate is rotatably connected to the rotating rod, and a baffle is fixedly arranged on the connecting plate;

[0016] The baffles are triangular in structure, and any two adjacent baffles are in contact and connected, and the two ends of each baffle are in contact and connected with the mounting top ring and the mounting bottom ring respectively, and a plurality of the baffles, the mounting top ring, the mounting bottom ring and the bearing mesh plate form a filling cavity, and the filling cavity is filled with gel particles;

[0017] The gel cavity is formed by the filling cavity and the through hole C being connected.

[0018] Preferably, the baffle has an inclined structure.

[0019] Preferably, the linkage assembly includes a rotating shaft, three sub-tooth rings and a mother tooth ring, the rotating shaft is rotatably arranged on the rotating groove, a gear is fixedly provided on the top of the rotating shaft, a clamping groove C is opened on the top of the gear, the bottom end of the connecting shaft passes through the clamping groove A and the clamping groove B in sequence and is fixedly connected to the bottom end of the clamping groove C, the three sub-tooth rings are respectively fixedly arranged on the top outer edge surfaces of the three mounting top rings, the gear is meshingly connected with the three sub-tooth rings, the mother tooth ring is rotatably arranged on the mounting cavity relative to the three sub-tooth rings, and the mother tooth ring is meshingly connected with the three sub-tooth rings.

[0020] Preferably, the adjustment assembly includes a clamping slider, a plurality of connecting rods, a movable ring and a sleeve, the clamping slider is sleeved on the connecting shaft relative to the clamping slide groove C, the two ends of the clamping slider are respectively clamped with the clamping slide groove B and the clamping slide groove C, the top of the clamping slider is clamped with the clamping slide groove A, wherein the height of the clamping slider is less than the height difference between the top of the clamping slide groove A and the bottom of the clamping slide groove B, the bottom end of the clamping slider is rotatably connected with a rotating ring C, and the plurality of connecting rods are arranged on the rotating shaft in a ring-shaped equidistant structure. The top end of the connecting rod is fixedly connected to the rotating ring C, the movable ring is movably arranged on the movable groove, the top end of the movable ring is fixedly connected to the bottom end of the connecting rod, the sleeve is fixedly arranged at the bottom end of the chromatography column, two plug holes A are provided on the sleeve, an adjusting rod is passed through the sleeve, the top end of the adjusting rod passes through the bottom end of the chromatography column and the bottom of the gel bottom column in turn, extends into the movable groove and is rotatably connected to the movable ring, and a plug hole B is provided at the bottom of the adjusting rod, and the plug hole B is plugged with the plug hole A through a latch unit.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] 1. The present invention designs the structure of a chromatography column, a rotating mechanism, a column A, a column B and a gel mechanism so that the two ends of the three gel cavities are respectively connected to the three through holes A and the three through holes B to form a mixed liquid filtration channel, a protease elution channel and a cleaning channel respectively. When the protease is extracted, the protease mixed liquid is filtered from the mixed liquid filtration channel, and the protease is left on the gel particles. The gel mechanism is driven to rotate by the rotating mechanism, so that the gel cavity originally in the mixed liquid filtration channel rotates to the protease elution channel and forms a new channel. The protease in the gel cavity is washed out by the eluent to obtain a protease eluent for subsequent protease analysis and purification. The gel cavity originally in the protease elution channel is eluted and rotated to the cleaning channel to form a new channel. The cleaning channel is cleaned with the cleaning solution. The gel cavity originally in the cleaning channel that has been cleaned rotates to the mixed liquid filtration channel to form a new channel, thereby achieving an automated continuous extraction effect of the protease, greatly saving time, and solving the technical problem of low extraction efficiency of the protease chromatography extraction equipment.

[0023] 2. The present invention adopts a structural design of the gel assembly so that the top ring is rotatably installed, the movable column A moves on the inclined guide groove, the position of the rotating rod changes, and the movable column B moves on the centripetal guide groove, causing a number of triangular baffles to move adaptively, resulting in a change in the size of the filling cavity surrounded by the baffles, the installation top ring, the installation bottom ring and the bearing mesh plate, and the overall shape of the gel particles filled in the filling cavity is adaptively changed. When the diameter of the overall shape of the gel particles increases, the height decreases, which is used to improve the filtration efficiency of the mixed liquid. When the diameter of the overall shape of the gel particles decreases, the height increases, which is used to improve the filtration effect of the mixed liquid. Compared with the traditional simple structure of filling gel particles in the chromatography column cavity, the height of the gel particles is easy to adjust, which saves the time for adjusting the height of the gel particles, further improves the extraction efficiency of the protease, and further solves the technical problem of low extraction efficiency of the protease chromatography extraction equipment.

[0024] 3. The present invention arranges the baffles in an inclined structure so that a plurality of baffles form a truncated cone-like structure that is larger at the top and smaller at the bottom. When the filling cavity is adjusted to be smaller, the baffles push the gel particles obliquely upward, which facilitates the adaptation of the overall shape of the gel particles filled in the filling cavity and is not easy to damage the gel particles, thereby preventing the gel particles from breaking and affecting the extraction effect of the protease.

[0025] 4. The present invention arranges the gel mechanism so that the adjusting rod can be raised and lowered, and is fixed by plugging the latch unit. The raising and lowering of the adjusting rod can drive the movable ring, the connecting rod, the rotating ring C and the clamping slider to rise and fall, and the height of the clamping slider is set to be smaller than the height difference between the top of the clamping slot A and the bottom of the clamping slot B, so that when the clamping slot C is clamped with the clamping slot B through the clamping slider, the gear rotates to drive the clamping slider to rotate, causing the gel top plate to rotate at the same time, so that the sub-toothed ring will not rotate by itself, and the entire gel mechanism as a whole (except for the partial adjusting components below the adjusting rod) rotates for the extraction of protease. When the segment rod is lifted and lowered until the movable ring moves to the top of the movable groove, the plug-in hole A located above is connected with the plug-in hole B and is fixed by the latch unit. At this time, the engaging slider rises and disengages from the engaging slide C, and the engaging slider enters the top of the engaging slide A, engaging the engaging slide A with the engaging slide B, so that the gel top plate and the gel bottom column cannot rotate. At this time, the gear drives the sub-tooth ring to rotate, causing the mother tooth ring to rotate relative to the installation cavity, thereby causing the three installation top rings to rotate at the same time, and adjusting the sizes of the three filling cavities at the same time, which is used to adjust the height of the gel layer formed by the gel particles according to the protease mixture before protease extraction. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is a schematic diagram of the overall structure of the present invention;

[0027] Figure 2 It is a schematic diagram of the overall cross-sectional structure of the present invention;

[0028] Figure 3 for Figure 2 A magnified schematic diagram of the local structure;

[0029] Figure 4 It is a schematic diagram of the partial structure disassembly of the present invention;

[0030] Figure 5 It is a structural schematic diagram of the column A of the present invention;

[0031] Figure 6 for Figure 4 A magnified schematic diagram of the local structure;

[0032] Figure 7 It is a schematic diagram of the explosion structure of the gel mechanism of the present invention;

[0033] Figure 8 for Figure 7 A magnified schematic diagram of the local structure;

[0034] Fig. 9 It is a schematic diagram of the cross-sectional structure of the gel mechanism of the present invention;

[0035] Fig.10 It is a schematic diagram of the enlarged structure of the local structure of the present invention;

[0036] Fig.11 It is a schematic diagram of the structure of the enclosure unit of the present invention.

[0037] Description of the numbers in the figure:

[0038] 1. Chromatographic column; 2. Rotating mechanism; 3. Column A; 4. Column B; 5. Gel mechanism;

[0039] 10. Installation cavity; 11. Flow guide pipe A; 12. Flow guide pipe B;

[0040] 21. Motor; 22. Coupling shaft;

[0041] 31. Through hole A; 32. Embedded groove A; 33. Sealing ring A; 34. Snap-fitting slide groove A;

[0042] 41. Through hole B; 42. Embossed groove B; 43. Sealing ring B;

[0043] 51. Gel bottom column; 52. Gel top plate; 53. Mounting hole; 54. Gel assembly; 55. Linkage assembly; 56. Adjustment assembly; 57. Connecting column; 58. Through hole C; 59. Snap-in slide groove B; 510. Rotating ring A; 511. Rotating groove; 512. Movable groove;

[0044] 540, swivel B; 541, top ring installation; 542, inclined guide groove; 543, bottom ring installation; 544, centripetal guide groove; 545, load-bearing mesh plate; 546, enclosure unit;

[0045] 551, rotating shaft; 552, gear; 553, engaging slideway C; 554, sub-gear ring; 555, main gear ring;

[0046] 560, swivel C; 561, clamping slider; 562, connecting rod; 563, movable ring; 564, sleeve; 565, plug-in hole A; 566, adjusting rod; 567, plug-in hole B; 568, latch unit;

[0047] 5461. Movable column A; 5462. Movable column B; 5463. Rotating rod; 5464. Connecting plate; 5465. Baffle. DETAILED DESCRIPTION

[0048] like Figures 1 to 11As shown, the present invention relates to a protease chromatography extraction device, comprising a chromatography column 1 and a rotating mechanism 2, wherein the interior of the chromatography column 1 is hollow to form an installation cavity 10, a column A3 is fixedly arranged on the top of the installation cavity 10, and three through holes A31 are provided on the column A3 in an annular structure with equal spacing, wherein the gap distance between any two through holes A31 is greater than the diameter of the through holes A31, three flow guide tubes A11 are connected to the top of the chromatography column 1, and the three flow guide tubes A11 are respectively connected to the tops of the three through holes A31, a column B4 is fixedly arranged on the bottom of the installation cavity 10, and three through holes B41 are provided on the column B4 in an annular structure with equal spacing, and the through holes B41 are adapted to the through holes A31, and three flow guide tubes B12 are connected to the bottom of the chromatography column 1, and the three flow guide tubes B12 are respectively connected to the bottoms of the three through holes B41, a gel mechanism 5 is arranged in the gap between the column A3 and the column B4, and the gel mechanism 5 is rotatably connected to the installation cavity 10;

[0049] The gel mechanism 5 includes three gel chambers, and the two ends of the three gel chambers are respectively connected to three through holes A31 and three through holes B41 to form a mixed liquid filtration channel, a protease elution channel and a cleaning channel;

[0050] The rotating mechanism 2 is arranged at the top of the chromatography column 1, and the output end of the rotating mechanism 2 passes through the chromatography column 1 and the column A3 in sequence and is connected to the gel mechanism 5. The present invention prevents the through hole A31 from being connected to two gel chambers at the same time by setting the gap between any two through holes A31 to be greater than the diameter of the through hole A31, so as to prevent the protease mixture, the protease eluate and the cleaning channel from being mixed.

[0051] In an embodiment of the present invention, the rotating mechanism 2 includes a motor 21 and a coupling 22, the motor 21 is fixedly mounted at the top of the chromatography column 1, the coupling 22 is passed through the center of the column A3, and the coupling 22 is rotationally connected to the column A3, and the top of the coupling 22 passes through the top of the chromatography column 1 and is fixedly connected to the output shaft of the motor 21. The present invention drives the coupling 22 to rotate by setting the motor 21.

[0052] In the embodiment of the present invention, three embedding grooves A32 are provided at the bottom of the column A3 relative to the three through holes A31, and a sealing ring A33 is embedded in the embedding groove A32. The bottom end of the sealing ring A33 is in contact with the top end of the gel mechanism 5, and a clamping groove A34 is provided at the center of the bottom end of the column A3.

[0053] Three embedding grooves B42 are provided at the top of the column B4 at positions corresponding to the three through holes B41. Sealing rings B43 are embedded in the embedding grooves B42. The sealing rings B43 are in contact with and connected to the bottom of the gel mechanism 5.

[0054] Among them, the sealing ring A33 and the sealing ring B43 are both made of polytetrafluoroethylene. The present invention, through the setting of the sealing ring A33 and the sealing ring B43, makes the rotating mechanism 2 contact with the cylinder A3 and the cylinder B4 respectively through the sealing ring A33 and the sealing ring B43, prevents the mixed liquid filtering channel, the protease elution channel and the cleaning channel from mixing liquid, and sets the sealing ring A33 and the sealing ring B43 to be made of polytetrafluoroethylene, which is a known sealing ring material with the smallest friction coefficient, and the friction coefficient is very low, so that the gel mechanism 5 is easy to rotate, and the seal is not easy to wear, thereby extending its service life, and polytetrafluoroethylene can withstand the erosion of most chemical substances such as strong acids, strong alkalis, oxidants and reducing agents, so that it can maintain stable performance when the mixed liquid and the elution liquid are in contact.

[0055] In the embodiment of the present invention, the gel mechanism 5 includes a gel bottom column 51 and a gel top plate 52. The gel bottom column 51 is rotatably connected to the installation cavity 10 through a plurality of rotating rings A510. A rotating groove 511 is provided at the center position of the top end of the gel bottom column 51. A movable groove 512 is provided at the bottom end of the rotating groove 511. A mounting hole 53 is provided on the gel bottom column 51 at a position corresponding to the three through holes B41. A gel assembly 54 is arranged on the mounting hole 53. The three gel assemblies 54 are linked by a linkage assembly 55. An adjustment assembly 56 is arranged on the linkage assembly 55. The gel top plate 52 is arranged above the gel bottom column 51. The gel top plate 52 and the gel bottom column 51 are fixedly connected by three connecting columns 57 arranged in a ring-shaped equidistant structure. The top end of the gel top plate 52 is in contact with the sealing ring A33. Three through holes C58 are provided on the gel top plate 52 at positions corresponding to the three through holes A31. A snap-in sliding groove B59 is provided at the center position of the gel top plate 52.

[0056] In an embodiment of the present invention, the gel assembly 54 includes an installation top ring 541 and an installation bottom ring 543. The installation top ring 541 is rotatably arranged at the top of the installation hole 53. The installation top ring 541 is rotatably connected to the gel top plate 52 through a swivel B540. The bottom end of the installation top ring 541 is provided with a plurality of inclined guide grooves 542 in an annular structure with equal spacing. The installation bottom ring 543 is fixedly arranged at the bottom end of the installation hole 53. The bottom end of the installation bottom ring 543 is in contact with the sealing ring B43. The installation bottom ring 543 is provided with a plurality of centripetal guide grooves 544 in an annular structure with equal spacing relative to the plurality of inclined guide grooves 542. A load-bearing mesh plate 545 is fixedly installed at the top of the ring cavity of the installation bottom ring 543. The centripetal guide groove 544 is movably connected to the inclined guide groove 542 through a blocking unit 546.

[0057] The enclosure unit 546 includes a movable column A5461 and a movable column B5462. The movable column A5461 is movably arranged on the oblique guide groove 542, and the movable column B5462 is movably arranged on the centripetal guide groove 544. The movable column A5461 and the movable column B5462 are rotatably connected by a rotating rod 5463. A connecting plate 5464 is rotatably connected to the rotating rod 5463, and a baffle 5465 is fixedly arranged on the connecting plate 5464.

[0058] The baffles 5465 are in a triangular structure, and any two adjacent baffles 5465 are in contact and connected, and the two ends of each baffle 5465 are in contact and connected with the mounting top ring 541 and the mounting bottom ring 543 respectively. A plurality of baffles 5465, the mounting top ring 541, the mounting bottom ring 543 and the bearing mesh plate 545 form a filling cavity, and the filling cavity is filled with gel particles.

[0059] The gel cavity is connected by a filling cavity and a through hole C58. The present invention, through the structural design of the gel assembly 54, makes the top ring 541 rotatably installed, the movable column A5461 moves on the inclined guide groove 542, the position of the rotating rod 5463 changes, and the movable column B5462 moves on the centripetal guide groove 544, so that a plurality of triangular baffles 5465 are adaptively movable, so that the size of the filling cavity surrounded by a plurality of baffles 5465, the top ring 541, the bottom ring 543 and the bearing mesh plate 545 changes, and the overall shape of the gel particles filled in the filling cavity changes adaptively. When the diameter of the overall shape of the gel particles increases, the height decreases, which is used to improve the filtration efficiency of the mixed solution. When the diameter of the overall shape of the gel particles decreases, the height increases, which is used to improve the filtration effect of the mixed solution. Compared with the traditional simple structure of filling gel particles in the chromatography column cavity, the height of the gel particles is convenient to adjust, which saves the time of adjusting the height of the gel particles, and further improves the extraction efficiency of the protease, thereby further solving the technical problem of low extraction efficiency of the protease chromatography extraction equipment.

[0060] In the embodiment of the present invention, the baffle 5465 is in an inclined structure. The present invention arranges the baffle 5465 in an inclined structure, so that a plurality of baffles 5465 form a truncated cone structure with a large top and a small bottom, so that when the filling cavity is adjusted to be smaller, the baffle 5465 pushes the gel particles obliquely upward, so that the overall shape of the gel particles filled in the filling cavity can be adapted to change, and the gel particles are not easily damaged, so as to prevent the gel particles from breaking and affecting the extraction effect of the protease.

[0061] In an embodiment of the present invention, the linkage assembly 55 includes a rotating shaft 551, three sub-tooth rings 554 and a mother tooth ring 555. The rotating shaft 551 is rotatably arranged on the rotating groove 511. A gear 552 is fixedly arranged on the top of the rotating shaft 551. A snap-in groove C553 is opened on the top of the gear 552. The bottom end of the connecting shaft 22 passes through the snap-in groove A34 and the snap-in groove B59 in sequence and is fixedly connected to the bottom end of the snap-in groove C553. The three sub-tooth rings 554 are respectively fixed on the top outer edge surfaces of the three mounting top rings 541. The gear 552 is meshed with the three sub-tooth rings 554. The mother tooth ring 555 is rotatably arranged on the mounting cavity 10 relative to the three sub-tooth rings 554. The mother tooth ring 555 is meshed with the three sub-tooth rings 554. The present invention designs the linkage assembly 55 so that the rotation of the connecting shaft 22 can drive the gear 552 to rotate, and by setting a mother gear ring 555 to mesh with the three sub-gear rings 554, the three sub-gear rings 554 rotate in the same state, so that when one of the sub-gear rings 554 is rotated, the three mounting top rings 541 rotate simultaneously.

[0062] In the embodiment of the present invention, the adjustment component 56 includes a clamping slider 561, a plurality of connecting rods 562, a movable ring 563 and a sleeve 564. The clamping slider 561 is sleeved on the connecting shaft 22 relative to the clamping slide groove C553. The two ends of the clamping slider 561 are respectively clamped with the clamping slide groove B59 and the clamping slide groove C553. The top of the clamping slider 561 is clamped with the clamping slide groove A34. The height of the clamping slider 561 is less than the height difference between the top of the clamping slide groove A34 and the bottom of the clamping slide groove B59. The bottom end of the clamping slider 561 is rotatably connected with a rotating ring C560. The plurality of connecting rods 562 are arranged in a ring-shaped structure with equal spacing through the rotating shaft 55 1, the top of the connecting rod 562 is fixedly connected to the rotating ring C560, the movable ring 563 is movably arranged on the movable groove 512, the top of the movable ring 563 is fixedly connected to the bottom of the connecting rod 562, the sleeve 564 is fixedly arranged at the bottom end of the chromatography column 1, two plug holes A565 are arranged on the sleeve 564, and an adjusting rod 566 is passed through the sleeve 564. The top of the adjusting rod 566 passes through the bottom end of the chromatography column 1 and the bottom of the gel bottom column 51 in sequence and extends into the movable groove 512 and is rotatably connected to the movable ring 563. The bottom of the adjusting rod 566 is provided with a plug hole B567, and the plug hole B567 is plugged with the plug hole A565 through a latch unit 568. The present invention adopts the above arrangement, so that the adjusting rod 566 can be raised and lowered, and is fixed by plugging the latch unit 568. The raising and lowering of the adjusting rod 566 can drive the movable ring 563, the connecting rod 562, the rotating ring C560 and the clamping slider 561 to rise and fall, and the height of the clamping slider 561 is set to be smaller than the height difference between the top of the clamping slot A34 and the bottom of the clamping slot B59, so that when the clamping slot C553 is clamped with the clamping slot B59 through the clamping slider 561, the gear 552 rotates to drive the clamping slider 561 to rotate, so that the gel top plate 52 rotates at the same time, so that the sub-toothed ring 554 does not rotate by itself, and the entire gel mechanism 5 rotates as a whole (except for the part of the adjusting component 56 below the adjusting rod 566) for the extraction of protease. When the segment rod 566 is lifted and lowered until the movable ring 563 moves to the top of the movable groove 512, the plug-in hole A565 located above is connected with the plug-in hole B567, and is plugged and fixed by the latch unit 568. At this time, the engaging slider 561 rises and disengages from the engaging slide C553, and the engaging slider 561 enters the top of the engaging slide A34, engaging the engaging slide A with the engaging slide B59, so that the gel top plate 52 and the gel bottom column 51 cannot rotate. At this time, the gear 552 drives the sub-tooth ring 554 to rotate, causing the mother tooth ring 555 to rotate relative to the installation cavity 10, thereby causing the three installation top rings 541 to rotate at the same time, and adjusting the sizes of the three filling cavities at the same time, which is used to adjust the height of the gel layer formed by the gel particles according to the protease mixture before protease extraction.

[0063] Working principle: This embodiment provides a protease chromatography extraction device. When in use, the height of the gel layer formed by the gel particles is adjusted according to the protease mixture. When the hand-held adjusting rod 566 rises to the movable ring 563 and moves to the top of the movable groove 512, the plug-in hole A565 located above is connected with the plug-in hole B567, and is plugged and fixed by the latch unit 568. At this time, the clamping slider 561 rises and disengages from the clamping slide C553, and the clamping slider 561 enters the top of the clamping slide A34, clamping the clamping slide A with the clamping slide B59, so that the gel top plate 52 and the gel bottom column 51 cannot rotate. The motor 21 is controlled by the external control mechanism to drive the connecting shaft 22 to rotate. When the gear 552 drives the sub-gear ring 554 to rotate, the mother gear ring 555 rotates relative to the installation cavity 10, so that the three installation top rings 541 rotate at the same time. The rotation of the installation top ring 541 drives the movable column A5461 to move on the inclined guide groove 542, the position of the rotating rod 5463 changes, and the movable column B5462 moves on the centripetal guide groove 544, causing a number of triangular baffles 5465 to move adaptively, causing the size of the filling cavity surrounded by the baffles 5465, the installation top ring 541, the installation bottom ring 543 and the bearing mesh plate 545 to change. The baffle 5465 pushes the gel particles obliquely upward, and the overall shape of the gel particles filled in the filling cavity changes adaptively until the adjustment is completed;

[0064] When the hand-held adjustment rod 566 is lowered until the movable ring 563 moves to the bottom of the movable groove 512, the plug-in hole A565 located below is connected with the plug-in hole B567, and is plugged and fixed by the latch unit 568, and the clamping slide C553 is clamped with the clamping slide B59 through the clamping slider 561. At this time, the motor 21 is controlled by the external control mechanism to drive the connecting shaft 22 to rotate, and the gear 552 rotates to drive the clamping slider 561 to rotate, causing the gel top plate 52 to rotate at the same time, so that the sub-toothed ring 554 will not rotate by itself, and the entire gel mechanism 5 rotates as a whole (except for the partial adjustment 56 components below the adjustment rod 566);

[0065] The protease mixture is filtered through the mixed liquid filtration channel, and the protease remains on the gel particles. The rotating mechanism 2 drives the gel mechanism 5 to rotate, so that the gel cavity originally in the mixed liquid filtration channel rotates to the protease elution channel and forms a new channel. The protease in the gel cavity is washed out by the eluent to obtain a protease eluent for subsequent analysis and purification. The gel cavity originally in the protease elution channel is rotated to the cleaning channel after elution and forms a new channel. The cleaning channel is cleaned with the cleaning liquid. The gel cavity originally in the cleaning channel that has been cleaned rotates to the mixed liquid filtration channel to form a new channel, thereby realizing the automated continuous extraction effect of the protease, greatly saving time, and solving the technical problem of low extraction efficiency of the protease chromatography extraction equipment.

[0066] The embodiments of the present invention disclose preferred embodiments, but are not limited thereto. A person skilled in the art can easily understand the spirit of the present invention based on the above embodiments and make different extensions and changes. However, as long as they do not deviate from the spirit of the present invention, they are all within the protection scope of the present invention.

Claims

1. A protease chromatography extraction device, characterized in that: The invention comprises a chromatography column (1) and a rotating mechanism (2), wherein the chromatography column (1) is hollow inside to form an installation cavity (10), a column A (3) is fixedly arranged on the top of the installation cavity (10), and three through holes A (31) are formed on the column A (3) in an annular structure with equal spacing, wherein the gap between any two of the through holes A (31) is greater than the diameter of the through holes A (31), and three flow guide tubes A (11) are connected to the top of the chromatography column (1), and the three flow guide tubes A (11) are respectively connected to the tops of the three through holes A (31). A column B (4) is fixedly provided at the bottom end of the installation cavity (10), and three through holes B (41) are formed in an annular structure with equal spacing, and the through holes B (41) are adapted to the through holes A (31). Three flow guide tubes B (12) are connected to the bottom end of the chromatography column (1), and the three flow guide tubes B (12) are respectively connected to the bottom ends of the three through holes B (41). A gel mechanism (5) is arranged in the gap between the column A (3) and the column B (4), and the gel mechanism (5) is rotatably connected to the installation cavity (10); The gel mechanism (5) comprises three gel chambers, and two ends of the three gel chambers are respectively connected to the three through holes A (31) and the three through holes B (41) to form a mixed liquid filtration channel, a protease elution channel and a cleaning channel respectively; The rotating mechanism (2) is arranged at the top of the chromatography column (1), and the output end of the rotating mechanism (2) passes through the chromatography column (1) and the column body A (3) in sequence and is connected to the gel mechanism (5).

2. The protease chromatography extraction equipment according to claim 1, characterized in that: The rotating mechanism (2) comprises a motor (21) and a connecting shaft (22), wherein the motor (21) is fixedly mounted on the top end of the chromatography column (1), the connecting shaft (22) is passed through the center of the column A (3), and the connecting shaft (22) is rotationally connected to the column A (3), and the top end of the connecting shaft (22) passes through the top end of the chromatography column (1) and is fixedly connected to the output shaft of the motor (21).

3. The protease chromatography extraction equipment according to claim 2, characterized in that: The bottom end of the column A (3) is provided with three embedding grooves A (32) at positions relative to the three through holes A (31), and a sealing ring A (33) is embedded in the embedding groove A (32). The bottom end of the sealing ring A (33) is in contact with and connected to the top end of the gel mechanism (5), and a snap-in sliding groove A (34) is provided at the center position of the bottom end of the column A (3).

4. The protease chromatography extraction equipment according to claim 3, characterized in that: Three embedding grooves B (42) are formed at the top of the column B (4) at positions corresponding to the three through holes B (41), and sealing rings B (43) are embedded in the embedding grooves B (42). The sealing rings B (43) are in contact with and connected to the bottom end of the gel mechanism (5); Wherein, the sealing ring A (33) and the sealing ring B (43) are both made of polytetrafluoroethylene.

5. The protease chromatography extraction equipment according to claim 4, characterized in that: The gel mechanism (5) comprises a gel bottom column (51) and a gel top plate (52), wherein the gel bottom column (51) is rotatably connected to the mounting cavity (10) via a plurality of swivels A (510), a swivel groove (511) is provided at the center of the top of the gel bottom column (51), and a movable groove (512) is provided at the bottom of the swivel groove (511), and a mounting hole (53) is provided on the gel bottom column (51) at a position corresponding to the three through holes B (41), and a gel assembly (54) is arranged on the mounting hole (53), and the three gel assemblies (54) are connected via a linkage assembly (5 5) linkage, wherein an adjustment component (56) is arranged on the linkage component (55), the gel top plate (52) is arranged above the gel bottom column (51), the gel top plate (52) and the gel bottom column (51) are fixedly connected by three connecting columns (57) arranged in a ring-shaped equidistant structure, the top end of the gel top plate (52) is in contact with the sealing ring A (33), three through holes C (58) are provided on the gel top plate (52) at positions corresponding to the three through holes A (31), and a snap-in groove B (59) is provided at the center of the gel top plate (52).

6. The protease chromatography extraction equipment according to claim 5, characterized in that: The gel assembly (54) comprises a mounting top ring (541) and a mounting bottom ring (543), wherein the mounting top ring (541) is rotatably arranged at the top end of the mounting hole (53), the mounting top ring (541) is rotatably connected to the gel top plate (52) via a swivel B (540), the bottom end of the mounting top ring (541) is provided with a plurality of inclined guide grooves (542) in an annular structure with equal spacing, the mounting bottom ring (543) is fixedly arranged at the bottom end of the mounting hole (53), the bottom end of the mounting bottom ring (543) is in contact with the sealing ring B (43), the mounting bottom ring (543) is provided with a plurality of centripetal guide grooves (544) in an annular structure with equal spacing relative to the number of the inclined guide grooves (542), a bearing mesh plate (545) is fixedly installed at the top end of the ring cavity of the mounting bottom ring (543), and the centripetal guide groove (544) is movably connected to the inclined guide groove (542) via a blocking unit (546).

7. The protease chromatography extraction equipment according to claim 6, characterized in that: The enclosure unit (546) comprises a movable column A (5461) and a movable column B (5462), wherein the movable column A (5461) is movably disposed on the oblique guide groove (542), and the movable column B (5462) is movably disposed on the centripetal guide groove (544), and the movable column A (5461) and the movable column B (5462) are rotatably connected via a rotating rod (5463), a connecting plate (5464) is rotatably connected to the rotating rod (5463), and a baffle (5465) is fixedly disposed on the connecting plate (5464); The baffles (5465) are triangular in structure, and any two adjacent baffles (5465) are in contact and connected, and two ends of each baffle (5465) are in contact and connected with the mounting top ring (541) and the mounting bottom ring (543) respectively, and a plurality of baffles (5465), the mounting top ring (541), the mounting bottom ring (543) and the bearing mesh plate (545) form a filling cavity, and the filling cavity is filled with gel particles; The gel cavity is formed by the filling cavity and the through hole C (58) being connected.

8. The protease chromatography extraction equipment according to claim 7, characterized in that: The baffle (5465) has an inclined structure.

9. The protease chromatography extraction equipment according to claim 8, characterized in that: The linkage assembly (55) comprises a rotating shaft (551), three sub-toothed rings (554) and a main toothed ring (555); the rotating shaft (551) is rotatably mounted on the rotating groove (511); a gear (552) is fixedly mounted on the top of the rotating shaft (551); a clamping groove C (553) is formed on the top of the gear (552); the bottom end of the connecting shaft (22) passes through the clamping groove A (34) and the clamping groove B (59) in sequence and is connected to the clamping groove A (34) and the clamping groove B (59). The bottom end of the clamping groove C (553) is fixedly connected, the three sub-tooth rings (554) are respectively fixedly arranged on the top outer edge surfaces of the three mounting top rings (541), the gear (552) is meshingly connected with the three sub-tooth rings (554), the mother tooth ring (555) is rotatably arranged on the mounting cavity (10) relative to the three sub-tooth rings (554), and the mother tooth ring (555) is meshingly connected with the three sub-tooth rings (554).

10. The protease chromatography extraction equipment according to claim 9, characterized in that: The adjusting assembly (56) comprises a clamping slider (561), a plurality of connecting rods (562), a movable ring (563) and a sleeve (564); the clamping slider (561) is sleeved on the connecting shaft (22) relative to the clamping groove C (553); the two ends of the clamping slider (561) are respectively clamped with the clamping groove B (59) and the clamping groove C (553); the top of the clamping slider (561) is clamped with the clamping groove A (34); wherein the height of the clamping slider (561) is smaller than the height difference between the top of the clamping groove A (34) and the bottom of the clamping groove B (59); the bottom of the clamping slider (561) is rotatably connected with a rotating ring C (560); the plurality of connecting rods (562) are arranged on the rotating shaft (551) in an annular structure with equal spacing; the ...; the connecting rods (562) are arranged on the rotating shaft (551) in an annular structure; the connecting rods (562) are arranged on the rotating shaft (551) in annular structure; the connecting rods (562) are arranged on the rotating shaft (551) in annular structure; the connecting rods (562) are arranged ) is fixedly connected to the rotating ring C (560), the movable ring (563) is movably arranged on the movable groove (512), the top of the movable ring (563) is fixedly connected to the bottom of the connecting rod (562), the sleeve (564) is fixedly arranged at the bottom of the chromatography column (1), two plug holes A (565) are provided on the sleeve (564), an adjusting rod (566) is passed through the sleeve (564), the top of the adjusting rod (566) passes through the bottom of the chromatography column (1) and the bottom of the gel bottom column (51) in turn, extends into the movable groove (512) and is rotatably connected to the movable ring (563), the bottom of the adjusting rod (566) is provided with a plug hole B (567), the plug hole B (567) and the plug hole A (565) are plugged and matched through a latch unit (568).

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