CN217340171U - Chromatography system for urokinase production - Google Patents

Abstract

The utility model discloses a chromatography system for urokinase production, which comprises a chromatography column, wherein the chromatography column comprises a hollow elution cylinder with both upper and lower openings, an upper end cap assembly, a lower end cap assembly and a filter screen arranged in the lower end cap assembly. The cover assembly and the lower end cover assembly are respectively arranged on the top and the bottom of the elution cylinder; the upper end cover assembly is provided with a flushing port and a stationary phase inlet, and the flushing port is respectively connected with a water pipe, an eluent pipe and a dissolving liquid pipe through a conveying main pipe , the inlet ends of the water pipe, the eluent pipe and the liberation liquid pipe are respectively connected with the water cylinder, the eluent cylinder and the liberation liquid cylinder; the lower end cover assembly is provided with a liquid discharge port, and the liquid discharge port passes through a suction pump and a three-way valve respectively. A waste liquid pipe and a dissolving liquid collecting pipe are connected, and the outlet ends of the waste liquid pipe and the dissolving liquid collecting pipe are respectively connected with a waste liquid tank and a dissolving liquid collecting tank. The whole elution process uses automated equipment, which saves labor, and the low temperature environment is controlled during the process to ensure the activity and quality of urokinase.

Description

Chromatography System for Urokinase Production

technical field

The utility model belongs to the technical field of pharmaceutical equipment, in particular to a chromatography system for urokinase production.

Background technique

Urokinase refers to a protease isolated and extracted from healthy human urine. It acts directly on the endogenous fibrinolysis system and can catalyze the cleavage of plasminogen into plasmin. Plasmin can not only degrade fibrin clots, but also It can also degrade fibrinogen, coagulation factor V and coagulation factor VIII in the blood circulation, thereby exerting a thrombolytic effect, increasing the activity of vascular ADP enzyme, inhibiting ADP-induced platelet aggregation, and preventing thrombosis. Clinically, it is mainly used for the treatment of acute myocardial infarction, acute cerebral thrombosis, cerebrovascular embolism, thrombotic occlusive disease, pulmonary embolism and central retinal vein thrombosis.

Urokinase is prepared by reprocessing crude urokinase, and the crude urokinase needs to be adsorbed on silica gel during the extraction process. After adsorption, it is put into a chromatography column, and the urokinase is removed from the silica gel by the eluent. It is divided into the following steps: fresh urine collection - impurity removal - dilute hydrochloric acid to adjust the pH of urine to acidity - silica gel adsorption - cleaning - elution - ammonium sulfate precipitation and filtration - siphon drying The crude product is obtained - silica gel treatment and reuse. Because urokinase in urine is excreted from the human body, it will be degraded in 8 hours at room temperature (<24°C), and when the temperature exceeds 30°C, urokinase will be degraded in 2 hours. Therefore, in the whole process, a low temperature environment must be maintained to avoid the degradation of urokinase. However, the existing urokinase production equipment is simple, lacks the low temperature environment required by urokinase, and the quality of urokinase cannot be guaranteed.

Secondly, since the mobile phase (including water, eluent and elution solution) in the chromatography column is settled down, filtered and collected by its own gravity, the sedimentation speed is too slow, resulting in a long elution time. The existing operation usually requires the staff to manually pressurize the top of the chromatographic column to accelerate the downward movement of the mobile phase. This operation increases the labor intensity of the staff, and the components in the eluent and elution solution will be sucked by the staff. have an impact on their health. In addition, in the traditional process, the mobile phase is usually added to the chromatography column manually, which requires manual observation of the liquid level state of the mobile phase in the chromatography column, which is time-consuming and labor-intensive.

Utility model content

The purpose of the utility model is to provide a chromatography system for urokinase production in order to solve the deficiencies of the prior art.

The utility model is implemented through the following technical solutions:

A chromatographic system for urokinase production, comprising a chromatographic column, the chromatographic column comprises a hollow elution cylinder with both upper and lower openings, an upper end cap assembly, a lower end cap assembly and a filter screen laterally arranged in the lower end cap assembly, the filter screen It can prevent the silica gel from flowing out from the bottom during the chromatography process. The upper end cap assembly and the lower end cap assembly are detachably arranged on the top and bottom of the elution cylinder respectively; the upper end cap assembly is provided with a flushing port and a stationary phase inlet. , the flushing port is respectively connected with a water pipe, an eluent pipe and a liberation liquid pipe through a conveying main pipe, and the inlet ends of the water pipe, the eluent pipe and the liberation liquid pipe are respectively connected with a water cylinder and an eluent cylinder. and the liberation liquid cylinder; the lower end cover assembly is provided with a liquid discharge port, and the liquid discharge port is respectively connected with a waste liquid pipe and a liberation liquid collection pipe through a suction pump and a three-way valve, and the waste liquid pipe and The outlet ends of the liberation liquid collection pipe are respectively connected with a waste liquid tank and a liberation liquid collection tank.

By arranging a suction pump under the chromatographic column, the utility model can speed up and adjust the moving speed of the mobile phase in the elution cylinder under the suction action of the suction pump, without the need for manual pressurization by the staff, thereby reducing the working time of the staff. Labor intensity. When in use, the silica gel adsorbed with urokinase enters the chromatography column through the stationary phase inlet, and the silica gel is blocked on the filter screen. First, the tap water from the water tank is passed into the chromatography column through the water pipe, and the silica gel is cleaned with tap water. The cleaned waste liquid flows into the waste liquid tank through the waste liquid pipe, and the cleaning is stopped when the color of the waste liquid is as clear as water. . Then pass the eluent from the eluent cartridge (ie: ammonia water and urokinase activity enhancer) into the chromatography column through the eluent tube, and use the eluent to elute the impurities on the silica gel. The liquid flows into the waste liquid tank through the waste liquid pipe. After the elution is completed, pass the elution solution (ie: ammonium chloride and urokinase activity enhancer) into the chromatography column through the elution solution tube, and use the elution solution to desorb the urokinase in the silica gel. , so that the urokinase is removed from the silica gel, and the urokinase after the removal flows into the removal solution collection tank with the removal solution through the removal solution collection tube to complete the elution of the crude urokinase product.

Optionally, the extracting liquid collection tank and the chromatography column are all arranged in the same incubator, the inner wall of the incubator is provided with a thermal insulation layer, the outer wall is provided with a controller, and the incubator is provided with a A refrigerator and a temperature sensor, the controller is respectively electrically connected with the temperature sensor, the refrigerator and the suction pump. The settings of the refrigerator, temperature sensor and controller create the low temperature environment needed to ensure the quality and activity of urokinase. When the temperature sensor detects that the temperature in the incubator is too high and needs to be cooled, it transmits the information to the controller, and the controller starts the refrigerator to cool down the incubator; when the temperature sensor detects that the temperature in the incubator is within a suitable range When inside, the signal is transmitted to the controller, and the controller turns off the refrigerator, so as to accurately control the temperature of the incubator.

Optionally, a sealing door is provided on one side of the incubator. The sealing door is provided for easy access to the chromatography column and elution collection tank.

Optionally, the bottoms of the water cylinder, the eluent cylinder and the liberation liquid cylinder are all higher than the top of the incubator; the inlet ends of the water pipe, the eluent tube and the liberation liquid pipe are respectively arranged on the water cylinder. , the bottom of the eluent cartridge and the elution cartridge. Gravity is used to make the water, eluent and elution solution in the water cylinder, eluent cylinder and elution solution cylinder flow into the chromatographic column, so as to avoid the electric energy loss caused by the suction pump.

Optionally, the water pipe, the eluent pipe and the liberation liquid pipe are all provided with a flow rate valve, and the flow rate valve and the three-way valve are respectively electrically connected to the controller. The opening and closing of the flow rate valve and the three-way valve are controlled by the controller. The flow rate valves are used to adjust the flow rates of water, eluent, and elution solution passing through the water pipe, eluent pipe, and elution liquid pipe, respectively.

Optionally, the water cylinder, the eluent cylinder, and the liberation liquid cylinder are all provided with a liquid level gauge, and the water cylinder, the eluent cylinder, and the liberation liquid cylinder are all provided with an alarm controlled by the liquid level gauge. . When the liquid level gauge detects that the liquid level is too high or too low, the alarm will start the alarm, so as to remind the staff to add or stop adding water, eluent or solution in time.

Optionally, the bottom of the chromatography column is communicated with one end of the shaped pipe, the other end of the U-shaped pipe is vertically upward and its height is not lower than the height of the top of the chromatography column, and the lower part and the upper part of the U-shaped pipe are respectively A low liquid level photoelectric sensor and a high liquid level photoelectric sensor are arranged; the low liquid level photoelectric sensor and the high liquid level photoelectric sensor are respectively connected with the controller in communication. The low-level photoelectric sensor and the high-level photoelectric sensor are used to monitor the lower limit liquid level and upper limit liquid level of the mobile phase in the chromatography column, respectively. When the sensor detects that the liquid level is higher than the upper limit liquid level, it transmits the signal to the controller, and the controller opens or closes the flow rate valve on the corresponding pipeline in time.

Optionally, the lower end cap assembly includes an upper ring member and a lower ring member, and the bottom of the lower ring member is sealed and provided with the liquid discharge port; the upper ring member and the lower ring member are both ring-shaped A number of vertical screw holes are arranged in the direction, and the vertical screw holes at the corresponding positions on the upper ring member and the lower ring member are fastened and connected by bolts; the filter screen is arranged between the upper ring member and the lower ring member. between. When the filter needs to be replaced after long-term use, the filter can be removed by loosening the bolts, and then the new filter can be placed between the upper ring and the lower ring, and then fastened with bolts. The structure can facilitate the replacement of the filter screen.

Optionally, the upper end cover assembly is also provided with a vent. The setting of the vent can prevent the negative pressure in the column and prevent the mobile phase from moving downward.

Compared with the prior art, the present utility model has the following beneficial effects: 1. The present utility model sets the incubator and the refrigerator and the temperature sensor in the incubator, and then places the chromatographic column and the extraction liquid collection tank in the incubator. It can be placed in a specific low temperature environment to avoid the degradation of urokinase and ensure the activity and quality of urokinase; 2. The utility model automates the entire elution process, reduces manual participation in the process, and avoids the need for staff 3. The utility model can speed up and adjust the moving speed of the mobile phase in the elution cylinder by arranging a suction pump under the chromatographic column, which is helpful for Improve the elution rate and shorten the production cycle; and no need for manual pressure by the staff, reducing the labor intensity of the staff.

Description of drawings

Fig. 1 is the overall structure schematic diagram of the present utility model;

FIG. 2 is a schematic structural diagram of the lower end cover assembly of the present invention.

In the figure: 1. Elution cartridge, 2. Upper end cap assembly, 3. Lower end cap assembly, 4. Filter screen, 5. Stationary phase inlet, 6. Main delivery pipe, 7. Water pipe, 8. Eluent pipe, 9. Elution tube, 10. Water tank, 11. Eluent drum, 12. Elution drum, 13. Suction pump, 14. Three-way valve, 15. Waste liquid pipe, 16. Elution liquid collection tube, 17. Waste liquid Tank, 18. Removal liquid collection tank, 19. Incubator, 20. Insulation layer, 21. Controller, 22. Refrigerator, 23. Temperature sensor, 24. Flow rate valve, 25. Liquid level gauge, 26. Alarm, 27. U-shaped tube, 28. Low liquid level photoelectric sensor, 29. High liquid level photoelectric sensor, 30. Upper ring, 31. Lower ring, 32. Screw hole, 33. Air vent.

Detailed ways

The present invention will be further described below with reference to the accompanying drawings and embodiments, but the protection scope of the present invention is not limited by the embodiments.

Example 1

As shown in Figure 1-2, the chromatography system for urokinase production includes a chromatography column, the chromatography column includes a hollow elution cylinder 1 with both upper and lower openings, an upper end cap assembly 2, a lower end cap assembly 3, and a laterally arranged lower end cap The filter screen 4 in the assembly 3, the filter screen 4 acts as a barrier to the silica gel, which can prevent the silica gel from flowing out from the bottom during the chromatography process. and the bottom; the upper end cap assembly 2 is provided with a flushing port and a stationary phase inlet 5, and the flushing port is respectively connected with a water pipe 7, an eluent pipe 8 and an eluent pipe 9 through a conveying main pipe 6, the water pipe 7, the eluent pipe 8 and the inlet end of the liberation liquid pipe 9 are respectively connected with a water cylinder 10, an eluent cylinder 11 and a liberation liquid cylinder 12; the lower end cover assembly 3 is provided with a liquid discharge port, and the liquid discharge port passes a suction pump 13 and a three-way valve 14. A waste liquid pipe 15 and a dissolving liquid collecting pipe 16 are respectively connected, and the outlet ends of the waste liquid pipe 15 and the dissolving liquid collecting pipe 16 are respectively connected with a waste liquid tank 17 and a dissolving liquid collecting tank 18 .

In the present invention, the suction pump 13 is arranged under the chromatographic column, and under the suction action of the suction pump 13, the moving speed of the mobile phase in the elution cylinder 1 can be accelerated and adjusted, and the manual pressure of the staff is not required, thereby reducing the need for manual pressure. The labor intensity of the staff. When in use, the silica gel adsorbed with urokinase enters the chromatography column through the stationary phase inlet 5 , and the silica gel is blocked on the filter screen 4 . First, the tap water from the water tank 10 is passed into the chromatography column through the water pipe 7, and the silica gel is washed with tap water. The washed waste liquid flows into the waste liquid tank 17 through the waste liquid pipe 15, and the color of the waste liquid is the same as that of water. Stop cleaning when cleaning. Then, the eluent (ie: ammonia water and urokinase activity enhancing agent) from the eluent cartridge 11 is passed into the chromatography column through the eluent tube 8, and the silica gel is eluted with the eluent to elute the impurities. The waste liquid flows into the waste liquid tank 17 through the waste liquid pipe 15 . After the elution is completed, the elution solution (ie: ammonium chloride and urokinase activity enhancer) from the elution solution cartridge 12 is passed into the chromatography column through the elution solution tube 9, and the elution solution is used to treat the urokinase in the silica gel. Carry out desorption to remove urokinase from the silica gel, and the removed urokinase flows into the removal solution collection tank 18 with the removal solution through the removal solution collection tube 16 to complete the elution of the crude urokinase product.

Wherein, the extracting liquid collection tank 18 and the chromatography column are all arranged in the same constant temperature box 19, the inner wall of the constant temperature box 19 is provided with a thermal insulation layer 20, the outer wall is arranged with a controller 21, and the constant temperature box 19 is arranged with a refrigerator 22 and The temperature sensor 23 and the controller 21 are respectively electrically connected with the temperature sensor 23 , the refrigerator 22 and the suction pump 13 . The settings of the refrigerator 22, temperature sensor 23 and controller 21 create the low temperature environment required to ensure the quality and activity of urokinase. When the temperature sensor 23 detects that the temperature in the incubator is too high and needs to be lowered, the information is transmitted to the controller 21, and the controller 21 starts the refrigerator 22 to cool down the incubator; when the temperature sensor 23 detects that the incubator 23 When the internal temperature is within an appropriate range, a signal is transmitted to the controller 21, and the controller 21 turns off the refrigerator 22, thereby accurately controlling the temperature of the incubator.

Among them, one side of the thermostatic box 19 is provided with a sealing door. The sealing door is provided for easy access to the chromatography column and elution collection tank.

Wherein, the bottoms of the water cylinder 10, the eluent cylinder 11 and the eluting liquid cylinder 12 are all higher than the top of the incubator; The bottom of the cylinder 11 and the stripping liquid cylinder 12. Utilize the action of gravity to make the water, eluent and eluate in the water cylinder 10, the eluent cylinder 11 and the eluting liquid cylinder 12 flow into the chromatographic column, so as to avoid the electric energy loss caused by the suction of the suction pump 13.

Wherein, the water pipe 7 , the eluent pipe 8 and the stripping liquid pipe 9 are all provided with a flow rate valve 24 , and the flow rate valve 24 and the three-way valve 14 are respectively electrically connected to the controller 21 . The opening and closing of the flow rate valve 24 and the three-way valve 14 are controlled by the controller 21 . The flow rate valve 24 is used to adjust the flow rates of the water, the eluent and the elution solution passing through the water pipe 7 , the eluent pipe 8 and the elution liquid pipe 9 respectively.

Among them, the water tank 10, the eluent tank 11, and the removal liquid tank 12 are all provided with a liquid level gauge 25, and the water tank 10, the eluent liquid tank 11, and the removal liquid tank 12 are all provided with an alarm controlled by the liquid level meter 25. 26. When the liquid level gauge 25 detects that the liquid level is too high or too low, the alarm 26 starts an alarm, so as to remind the staff to add or stop adding water, eluent or solution in time.

Wherein, the bottom of the chromatography column is communicated with one end of the U-shaped pipe 27, the other end of the U-shaped pipe 27 is vertically upward and its height is not lower than the height of the top of the chromatography column, and the lower part and the upper part of the U-shaped pipe 27 are respectively provided with low liquid The level photoelectric sensor 28 and the high liquid level photoelectric sensor 29; the low liquid level photoelectric sensor 28 and the high liquid level photoelectric sensor 29 are respectively connected to the controller 21 for communication. The low-level photoelectric sensor 28 and the high-level photoelectric sensor 29 are respectively used to monitor the lower limit liquid level and the upper limit liquid level of the mobile phase in the chromatography column. When the liquid level photoelectric sensor 29 detects that the liquid level is higher than the upper limit liquid level, it transmits a signal to the controller 21, and the controller 21 opens or closes the flow rate valve 24 on the corresponding pipeline in time.

Wherein, the lower end cover assembly 3 includes an upper ring member 30 and a lower ring member 31, the bottom of the lower ring member 31 is sealed and is provided with a drain port; the upper ring member 30 and the lower ring member 31 are provided with a number of vertical screws in the circumferential direction. The holes 32 and the vertical screw holes 32 at corresponding positions on the upper ring member 30 and the lower ring member 31 are connected by bolts; the filter screen 4 is arranged between the upper ring member 30 and the lower ring member 31 . When the filter screen 4 needs to be replaced after long-term use, the filter screen 4 can be removed by loosening the bolts, and then the new filter screen 4 is placed between the upper ring 30 and the lower ring Can be solid. This structure can facilitate the replacement of the filter screen 4 .

Wherein, the upper end cover assembly 2 is also provided with a vent 33 . The arrangement of the vent 33 can prevent the negative pressure phenomenon in the chromatography column and prevent the downward movement of the mobile phase.

The present utility model is not limited to the above-mentioned optional embodiments, and anyone can draw other various forms of products under the inspiration of the present utility model. The technical solutions within the scope defined by the claims all fall within the protection scope of the present invention.

Claims (9)

1. The chromatographic system for urokinase production, comprising a chromatographic column, is characterized in that: the chromatographic column comprises a hollow elution cylinder (1), an upper end cap assembly (2), a lower end cap assembly (3) that are all open up and down ) and a filter screen (4) laterally arranged in the lower end cap assembly (3), the upper end cap assembly (2) and the lower end cap assembly (3) are detachably arranged on the top and the bottom of the elution cylinder (1) respectively. the bottom; the upper end cap assembly (2) is provided with a flushing port and a stationary phase inlet (5), and the flushing port is respectively connected with a water pipe (7) and an eluent pipe (8) through a conveying main pipe (6). ) and a solution pipe (9), the inlet ends of the water pipe (7), the eluent tube (8) and the solution tube (9) are respectively connected with a water cylinder (10), an eluent cylinder (11) and The release liquid cylinder (12); the lower end cover assembly (3) is provided with a liquid discharge port, and the liquid discharge port is respectively connected with a waste liquid pipe (15) through a suction pump (13) and a three-way valve (14). ) and a liberation liquid collection pipe (16), the waste liquid pipe (15) and the liberation liquid collection pipe (16) are respectively connected with a waste liquid tank (17) and a liberation liquid collection tank (18) at the outlet ends. 2. The chromatographic system for urokinase production according to claim 1 is characterized in that: the described solution collection tank (18) and the chromatographic column are all arranged in the same incubator (19), and the incubator is The inner wall of (19) is provided with a thermal insulation layer (20), the outer wall is provided with a controller (21), and a refrigerator (22) and a temperature sensor (23) are provided in the thermostatic box (19). The controller (21) is respectively electrically connected with the temperature sensor (23), the refrigerator (22) and the suction pump (13). 3. The chromatography system for urokinase production according to claim 2, wherein a sealing door is provided on one side of the incubator (19). 4. The chromatography system for urokinase production according to claim 3, characterized in that: the bottoms of the water cylinder (10), the eluent cylinder (11) and the elution cylinder (12) are all higher than the constant temperature the top of the tank (19); the inlet ends of the water pipe (7), the eluent pipe (8) and the elution liquid pipe (9) are respectively arranged on the water cylinder (10), the eluent cylinder (11) and the Disengage the bottom of the liquid cartridge (12). 5. The chromatographic system for urokinase production according to claim 2, wherein the water pipe (7), the eluent pipe (8) and the elution liquid pipe (9) are all provided with a flow rate valve ( 24), the flow rate valve (24) and the three-way valve (14) are respectively electrically connected to the controller (21). 6. The chromatographic system for urokinase production according to claim 1, characterized in that: the water cylinder (10), the eluent cylinder (11), and the extraction liquid cylinder (12) are all provided with a liquid level gauge (25), an alarm (26) controlled by the liquid level gauge (25) is provided on the water cylinder (10), the eluent cylinder (11), and the release liquid cylinder (12). 7. The chromatography system for urokinase production according to claim 2, wherein the bottom of the chromatography column is communicated with one end of the U-shaped pipe (27), and the other end of the U-shaped pipe (27) is vertically upward and Its height is not lower than the height of the top of the chromatography column, and the lower and upper parts of the U-shaped tube (27) are respectively provided with a low-level photoelectric sensor (28) and a high-level photoelectric sensor (29); the The low liquid level photoelectric sensor (28) and the high liquid level photoelectric sensor (29) are respectively connected in communication with the controller (21). 8. The chromatography system for urokinase production according to claim 1, wherein the lower end cap assembly (3) comprises an upper annular member (30) and a lower annular member (31), the lower annular member The bottom of (31) is sealed and provided with the liquid discharge port; the upper ring member (30) and the lower ring member (31) are provided with a number of vertical screw holes (32) in the circumferential direction. The vertical screw holes (32) at corresponding positions on the upper ring member (30) and the lower ring member (31) are fastened and connected by bolts; the filter screen (4) is arranged on the upper ring member (30) and the lower ring member (31). between the ring members (31). 9 . The chromatography system for urokinase production according to claim 1 , wherein the upper end cap assembly ( 2 ) is further provided with a vent ( 33 ). 10 .

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