Primary emulsion basic protein powder purification system and purification method thereof

Abstract

The colostrum basic protein powder purification system comprises a centrifugal pretreatment device, an affinity chromatography device and an ion exchange chromatography device, the centrifugal pretreatment device comprises a centrifugal tank body and a feeding pipe connected with the centrifugal tank body, and a centrifugal paddle is installed at the bottom of the centrifugal tank body; a filter is arranged in the center of the top of the centrifugal tank body; the outlet end of the filter is connected to affinity chromatography equipment; the affinity chromatography equipment comprises two affinity chromatography columns which are arranged in parallel, two-position three-way electromagnetic valves are respectively mounted at the liquid inlet ends and the liquid outlet ends of the affinity chromatography columns, and two inlets of the two-position three-way electromagnetic valves at the inlet ends of the affinity chromatography columns are respectively connected with the centrifugal pretreatment equipment and the flushing equipment; two outlets of the two-position three-way electromagnetic valve at the outlet end of the affinity chromatography column are respectively connected with the waste liquid recovery equipment and the ion exchange chromatography equipment; and an adjustable column bed mechanism is arranged in the affinity chromatography column. The defects in the prior art can be overcome, and the treatment efficiency of colostrum basic protein purification equipment is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of deep processing of dairy products, in particular to a colostrum alkaline protein powder purification system and a purification method thereof. Background Art

[0002] Colostrum basic protein (CBP) is a protein unique to colostrum that has multiple effects, including regulating intestinal function, enhancing immunity, and promoting bone health. Due to the extremely low content of CBP in colostrum, the presence of various impurity proteins in colostrum, and the competition between host cell proteins and nucleic acids for binding to the target protein on the chromatography medium, the purification of CBP is difficult. In the prior art, the purification of CBP is generally achieved using multi-stage chromatographic separation technology. However, existing chromatography equipment has limited separation efficiency, and to improve the purification efficiency of CBP, the investment in chromatography equipment must be increased, resulting in the production cost of CBP remaining high. Summary of the Invention

[0003] The technical problem to be solved by the present invention is to provide a colostrum alkaline protein powder purification system and a purification method thereof, which can solve the deficiencies of the existing technology and improve the processing efficiency of the colostrum alkaline protein purification equipment.

[0004] In order to solve the above technical problems, the technical solutions adopted by the present invention are as follows.

[0005] A colostrum alkaline protein powder purification system comprises a centrifugal pretreatment device, an affinity chromatography device and an ion exchange chromatography device. The centrifugal pretreatment device comprises a centrifugal tank body and a feed pipe connected to the centrifugal tank body, a centrifugal blade is installed at the bottom of the centrifugal tank body, a filter is provided at the top center of the centrifugal tank body, and the outlet end of the filter is connected to the affinity chromatography device; the affinity chromatography device comprises two affinity chromatography columns arranged in parallel, the liquid inlet end and the liquid outlet end of the affinity chromatography column are respectively installed with a two-position three-way solenoid valve for switching the two affinity chromatography columns, the two inlets of the two-position three-way solenoid valve at the inlet end of the affinity chromatography column are respectively connected to the centrifugal pretreatment device and the flushing device, and the two outlets of the two-position three-way solenoid valve at the outlet end of the affinity chromatography column are respectively connected to the waste liquid recovery device and the ion exchange chromatography device; an adjustable column bed mechanism is provided in the affinity chromatography column.

[0006] Preferably, a buffer tank is installed between the affinity chromatography device and the ion exchange chromatography device.

[0007] Preferably, a low-temperature refrigeration device is installed inside the centrifugal pretreatment equipment, affinity chromatography equipment, buffer tank and ion exchange chromatography equipment.

[0008] Preferably, the filter includes a shell, guide plates are installed at both ends of the shell, a filter membrane is installed between the two guide plates, a number of guide grooves are opened on the guide plates, and the guide grooves are symmetrically arranged with the center of the guide plate as the symmetry axis. The guide direction of the guide groove on the guide plate at the liquid inlet end of the filter is the same as the stirring direction of the centrifugal blade, and the guide direction of the guide groove on the guide plate at the liquid outlet end of the filter is opposite to the stirring direction of the centrifugal blade.

[0009] Preferably, an arc-shaped extension piece is symmetrically fixed on the guide groove of the guide plate at the liquid inlet end of the filter, close to the filter membrane. A swing blade is provided above the liquid inlet surface of the filter membrane. The swing blade is mounted on a connecting rod, and the connecting rod is driven to rotate by a stepper motor, which is integrated into the housing.

[0010] Preferably, the adjustable column bed mechanism includes a sleeve, both ends of the sleeve are respectively slidably engaged in a slide groove, the slide groove is fixed on the inner wall of the affinity chromatography column, a sealing ring is provided in the slide groove, a drive motor is installed on the inner wall of the affinity chromatography column, the drive motor drives the sleeve to rotate through a drive wheel, a central column is provided in the center of the sleeve, a plurality of partitions are provided on the outside of the central column, and chromatographic fillers are filled between adjacent partitions.

[0011] Preferably, an air bag is provided on the outside of the central column, and the air bag is connected to an external air pump through an air tube.

[0012] A purification method for the colostrum alkaline protein powder purification system comprises the following steps: continuously injecting colostrum protein lysate into a centrifugal tank through a feed pipe, centrifuging the colostrum protein lysate in the centrifugal tank under the rotation and agitation of centrifugal blades, and continuously passing through a filter under the action of injection pressure; filtrate passing through the filter enters an affinity chromatography device, an affinity chromatography column adsorbs the colostrum alkaline protein, two affinity chromatography columns are controlled by a two-position three-way solenoid valve, one affinity chromatography column is connected to the centrifugal tank for chromatographic filtration, and the other affinity chromatography column is connected to a flushing device for elution; the filtrate discharged after the chromatographic filtration enters a waste liquid recovery device, and the eluate containing the colostrum alkaline protein enters an ion exchange chromatography device, the ion exchange chromatography device adsorbs impurities, and purifies the colostrum alkaline protein.

[0013] The beneficial effect brought about by adopting the above technical solution is that the present invention improves the processing efficiency of the colostrum lysate in the purification equipment by improving the structure of the purification equipment. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is a structural diagram of a specific embodiment of the present invention.

[0015] Figure 2 It is a structural diagram of a centrifugal pretreatment device in a specific embodiment of the present invention.

[0016] Figure 3 It is a structural diagram of a filter in a specific embodiment of the present invention.

[0017] Figure 4 It is a partial enlarged view of the guide groove on the guide plate at the liquid inlet end of the filter close to the filter membrane in a specific embodiment of the present invention.

[0018] Figure 5 1 is a structural diagram of an affinity chromatography device in a specific embodiment of the present invention.

[0019] Figure 6 It is a structural diagram of an adjustable column bed mechanism in a specific embodiment of the present invention. DETAILED DESCRIPTION

[0020] Our R&D team has been deeply involved in colostrum alkaline protein purification for many years and filed a patent application for this process in 2024. During our refinement of the colostrum alkaline protein purification process, we discovered that the existing chromatography equipment had become a bottleneck for mass production of colostrum alkaline protein. To further improve production efficiency, we redeveloped the entire purification equipment suite, addressing many of the shortcomings of the existing chromatography equipment.

[0021] The entire purification system consists of a series of centrifugal pretreatment equipment 1, affinity chromatography equipment 2, buffer tank 3, and ion exchange chromatography equipment 4. To prevent the impact of temperature on protein molecules, a low-temperature refrigeration device 5 is used throughout the lysis process for low-temperature protection.

[0022] We employed a continuous feed process during the centrifugation and affinity chromatography process, significantly improving the efficiency of affinity chromatography. The centrifugal pretreatment apparatus 1 comprises a centrifuge tank 101 and a feed tube 102 connected to the tank. Centrifuge blades 103 are mounted at the bottom of the tank 101, and a filter 104 is located at the top center of the tank 101. The outlet of the filter 104 is connected to the affinity chromatography apparatus 2. We utilized feed injection pressure to continuously filter the lysate through the filter 104, allowing the filtrate to flow continuously into the affinity chromatography apparatus 2. The affinity chromatography device 2 includes two affinity chromatography columns 201 arranged in parallel. The liquid inlet and liquid outlet of the affinity chromatography column 201 are respectively installed with a two-position three-way solenoid valve 202 for switching between the two affinity chromatography columns 201. The two inlets of the two-position three-way solenoid valve 202 at the inlet end of the affinity chromatography column 201 are respectively connected to the centrifugal pretreatment device 1 and the flushing device 6. The two outlets of the two-position three-way solenoid valve 202 at the outlet end of the affinity chromatography column 201 are respectively connected to the waste liquid recovery device 7 and the ion exchange chromatography device 4. Under the control of the two-position three-way solenoid valve 202, the two affinity chromatography columns 201 are connected to the centrifugal tank 101 for chromatographic filtration, and the other affinity chromatography column 201 is connected to the flushing device 6 for elution. The eluate containing colostrum basic protein enters the buffer tank 3 and then enters the ion exchange chromatography device 4 to adsorb impurities, completing the purification of the colostrum basic protein.

[0023] Because the continuous centrifugal filtration process is used, the performance requirements for filter 104 are high, so we have redesigned filter 104. Filter 104 includes a housing 1041, with guide plates 1042 installed at both ends of housing 1041. A filter membrane 1043 is installed between the two guide plates 1042. A plurality of guide grooves 1044 are provided on the guide plates 1042. The guide grooves 1044 are symmetrically arranged with the center of the guide plates 1042 as the axis of symmetry. The guide grooves 1044 on the guide plate 1042 at the liquid inlet end of filter 104 have the same direction of flow as the stirring direction of centrifugal blades 103, while the guide grooves 1044 on the guide plate 1042 at the liquid outlet end of filter 104 have the opposite direction of flow as the stirring direction of centrifugal blades 103. The lysate enters the filter membrane 1043 through the guide grooves 1044 of the front guide plate 1042, reducing the influence of the guide grooves 1044 on the flow velocity. The filter membrane 1043 of the embodiment of the present invention is the filter membrane 1043 of the embodiment of the present invention.The ... By regularly changing the direction of swinging blades 1046, the high-speed flow of the lysate is utilized to clean the surface of filter membrane 1043, thereby extending the maintenance cycle of filter 104 and improving the equipment's uptime. Furthermore, when swinging blades 1046 are positioned between the two arc-shaped extensions 1045, the flow direction of the lysate can be adjusted by fine-tuning the angle of swinging blades 1046, thereby reducing turbulence generated by the lysate passing through guide grooves 1044.

[0024] The column bed of traditional affinity chromatography columns is installed once, and it is important to maintain the stability of the column bed during use to ensure that the packing remains undisturbed after homogenization and column pressing. However, these traditional affinity chromatography columns have slow elution rates, and if operated in parallel and continuously, five or six affinity chromatography columns are required to ensure uninterrupted chromatographic separation. To address this drawback of traditional affinity chromatography columns, we have designed a new affinity chromatography column with an adjustable column bed. This utilizes the centrifugal force generated by low-speed rotation to improve the elution efficiency of colostrum basic proteins attached to the packing. The sleeve 210 of the adjustable column bed mechanism 203 slides into the slide 211, which is fixed to the inner wall of the affinity chromatography column 201. A sealing ring 212 is provided in the slide 211, and a drive motor 213 is installed on the inner wall of the affinity chromatography column 201. The drive motor 213 drives the sleeve 210 to rotate via the drive wheel 214. A central column 215 is provided at the center of the sleeve 210, and a plurality of partitions 216 are provided on the outside of the central column 215. Chromatographic fillers are filled between adjacent partitions 216. When performing chromatographic separation, the sleeve 210 remains stationary. When elution is performed, the drive motor 213 drives the sleeve 210 to rotate; the rotation process adopts a smooth acceleration and deceleration method, and the absolute value of the acceleration is controlled at 100 rad / min. 2 The maximum speed is limited to 350 rad / min. Before activating the drive motor 213, the affinity chromatography column 201 is first eluted with the eluent. When the eluent flow rate stabilizes, the motor 213 is pneumatically driven. Finally, when the sleeve 210 stops rotating, the eluent injection is stopped. By packing the chromatography filler in sections, combined with speed control and eluent protection, disturbances to the filler caused by the rotation of the sleeve 210 can be effectively reduced.

[0025] In addition, an airbag 217 is disposed outside the central column 215 and connected to an external air pump 219 via an air tube 218. During chromatographic separation, airbag 217 is inflated to increase the packing density and improve the flow uniformity of the lysate. During elution, airbag 217 is deflated to reduce the mutual squeezing force between the packing and prepare for centrifugation. After centrifugation, airbag 217 is reinflated to quickly restore the packing density and prepare for the next round of chromatography.

[0026] The basic principles, main features, and advantages of the present invention are shown and described above. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The above embodiments and descriptions are merely illustrative of the principles of the present invention. Various changes and modifications may be made to the present invention without departing from the spirit and scope of the present invention. Such changes and modifications are intended to fall within the scope of the present invention. The scope of protection claimed in the present invention is defined by the appended claims and their equivalents.

Claims

1. A colostrum alkaline protein powder purification system, comprising a centrifugal pretreatment device (1), an affinity chromatography device (2) and an ion exchange chromatography device (4), characterized in that: The centrifugal pretreatment device (1) comprises a centrifugal tank body (101) and a feed pipe (102) connected to the centrifugal tank body (101), a centrifugal blade (103) is installed at the bottom of the centrifugal tank body (101), a filter (104) is arranged at the top center of the centrifugal tank body (101), and the outlet end of the filter (104) is connected to the affinity chromatography device (2); the affinity chromatography device (2) comprises two affinity chromatography columns (201) arranged in parallel, and the liquid inlet end and the liquid outlet end of the affinity chromatography column (201) are respectively installed with A two-position three-way solenoid valve (202) is used to realize the switching of the two affinity chromatography columns (201). The two inlets of the two-position three-way solenoid valve (202) at the inlet end of the affinity chromatography column (201) are respectively connected to the centrifugal pretreatment device (1) and the flushing device (6), and the two outlets of the two-position three-way solenoid valve (202) at the outlet end of the affinity chromatography column (201) are respectively connected to the waste liquid recovery device (7) and the ion exchange chromatography device (4); an adjustable column bed mechanism (203) is provided in the affinity chromatography column (201).

2. The colostrum alkaline protein powder purification system according to claim 1, characterized in that: A buffer tank (3) is installed between the affinity chromatography device and the ion exchange chromatography device.

3. The colostrum alkaline protein powder purification system according to claim 2, characterized in that: A low-temperature refrigeration device (5) is installed inside the centrifugal pretreatment equipment (1), affinity chromatography equipment (2), buffer tank (3) and ion exchange chromatography equipment (4).

4. The colostrum alkaline protein powder purification system according to claim 1, characterized in that: The filter (104) comprises a housing (1041), guide plates (1042) are installed at both ends of the housing (1041), a filter membrane (1043) is installed between the two guide plates (1042), a plurality of guide grooves (1044) are provided on the guide plates (1042), and the guide grooves (1044) are symmetrically arranged with the center of the guide plates (1042) as the symmetry axis. The guide direction of the guide grooves (1044) on the guide plate (1042) at the liquid inlet end of the filter (104) is the same as the stirring direction of the centrifugal blades (103), and the guide direction of the guide grooves (1044) on the guide plate (1042) at the liquid outlet end of the filter (104) is opposite to the stirring direction of the centrifugal blades (103).

5. The colostrum alkaline protein powder purification system according to claim 4, characterized in that: An arc-shaped extension piece (1045) is symmetrically fixed to a guide groove (1044) on a guide plate (1042) at the liquid inlet end of the filter (104) close to the filter membrane (1043). A swing blade (1046) is provided above the liquid inlet surface of the filter membrane (1043). The swing blade (1046) is mounted on a connecting rod (1047). The connecting rod (1047) is driven to rotate by a stepping motor (1048). The stepping motor (1048) is integrated into the housing (1041).

6. The colostrum alkaline protein powder purification system according to claim 1, characterized in that: The adjustable column bed mechanism (203) includes a sleeve (210), both ends of the sleeve (210) are respectively slidably engaged in a slide groove (211), the slide groove (211) is fixed on the inner wall of the affinity chromatography column (201), a sealing ring (212) is provided in the slide groove (211), a driving motor (213) is installed on the inner wall of the affinity chromatography column (201), the driving motor (213) drives the sleeve (210) to rotate through a driving wheel (214), a central column (215) is provided at the center of the sleeve (210), a plurality of partitions (216) are provided on the outside of the central column (215), and chromatographic fillers are filled between adjacent partitions (216).

7. The colostrum alkaline protein powder purification system according to claim 6, characterized in that: An air bag (217) is provided on the outside of the central column (215), and the air bag (217) is connected to an external air pump (219) through an air pipe (218).

8. A method for purifying colostrum alkaline protein powder according to any one of claims 1 to 7, characterized in that The following steps are involved: The colostrum protein lysate is continuously injected into the centrifugal tank (101) through the feed pipe (102), and the colostrum protein lysate is centrifuged in the centrifugal tank (101) under the rotation and stirring of the centrifugal blade (103), and continuously passes through the filter (104) under the action of the injection pressure; the filtrate passing through the filter (104) enters the affinity chromatography device (2), and the affinity chromatography column (201) adsorbs the colostrum alkaline protein. The two affinity chromatography columns (201) are controlled by the two-position three-way solenoid valve (202) to realize one affinity chromatography column (201) connected to the centrifugal tank (101) for chromatographic filtration, and the other affinity chromatography column (201) is connected to the flushing device (6) for elution. The filtrate discharged after the chromatographic filtration enters the waste liquid recovery device (7), and the eluate containing the colostrum alkaline protein enters the ion exchange chromatography device (4). The ion exchange chromatography device (4) adsorbs impurities and purifies the colostrum alkaline protein.

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