Device for extracting and purifying active protein

Abstract

The utility model provides an extraction and purification device for active protein. The extraction and purification device comprises a primary centrifugal kettle, a secondary centrifugal kettle and a tertiary centrifugal kettle, the interior of each centrifugal kettle is rotationally connected with a kettle core, and a filtrate space is formed between the kettle core and the centrifugal kettle shell; the kettle core comprises a twenty-six-surface outer framework made of steel, an inner framework with the same shape as the outer framework and an ultrafiltration membrane clamped between the outer framework and the inner framework; a filtrate outlet pipe is mounted on the centrifugal kettle shell, and the other end of the filtrate outlet pipe is communicated with the kettle core of the next stage of centrifugal kettle; and a concentrated outlet pipe communicated with the outside is arranged in the kettle core. The utility model provides a cascade centrifugal kettle, the kettle core arranged inside is provided with the ultrafiltration membrane of which the aperture is sequentially reduced, and the ultrafiltration membrane separates the filtrate and the concentrated solution of two separated products, so that the filtrate and the concentrated solution can be conveniently and respectively collected subsequently without worrying about the problem of secondary mixing, the operation difficulty is reduced, and the working efficiency is improved. The purity of protein purification is ensured, and the purification efficiency is improved.

Description

Technical Field

[0001] The utility model relates to the field of protein purification, in particular to an extraction and purification device for active protein. Background Art

[0002] Biological protein, also known as active protein, is the effective ingredient of many drugs or preparations. This type of protein is generally obtained from biological fermentation. The types of proteins produced by biological metabolism are mixed, so they need to be extracted and purified to extract the required active proteins. The methods currently used in the field include extraction, evaporation concentration or centrifugation, among which the extractant and heating evaporation will damage the active protein (chemical agents or high temperature will cause protein denaturation). Although the centrifugation method has little damage to the active protein, the separation product precipitate and filtrate obtained by the centrifugal equipment are currently in a centrifuge tube. The filtrate needs to be extracted from the top of the precipitate later. If the operation is not careful, the precipitate will be extracted together to cause mixing, resulting in low purity of protein purification. Utility Model Content

[0003] The purpose of the present application is to provide an extraction and purification device for active proteins, aiming to solve the above-mentioned problems in the prior art.

[0004] The embodiment of the present application provides an extraction and purification device for active protein, including a primary centrifugal pot, a secondary centrifugal pot and a tertiary centrifugal pot; each centrifugal pot is rotatably connected to a pot core inside, and a gap is left between the pot core and the outer shell of the centrifugal pot to form a filtrate space; the pot core includes an outer skeleton made of steel in the shape of a icosahedron, an inner skeleton with the same shape as the outer skeleton, and an ultrafiltration membrane clamped between the outer skeleton and the inner skeleton; the pore size of the ultrafiltration membrane in the primary centrifugal pot is the largest, and the pore size of the ultrafiltration membrane in the tertiary centrifugal pot is the smallest; a filtrate outlet pipe is installed on the outer shell of the centrifugal pot, the filtrate outlet pipe is connected to the filtrate space, and the other end of the filtrate outlet pipe is connected to the pot core of the next-stage centrifugal pot; a concentration outlet pipe connected to the outside world is arranged in the pot core; a motor is arranged at the bottom of the centrifugal pot, and the motor is transmission-connected to the bottom of the pot core.

[0005] Furthermore, the bottom of the centrifugal pot is installed on the base, and the motor is installed in the base; a lower connecting block is fixed on the outer frame at the lower end of the pot core; and the output shaft of the motor is drivingly connected to the lower connecting block.

[0006] Furthermore, a cylindrical body is arranged at the upper end of the base, the lower connecting block is located in the cylindrical body, and a lubricating pad is arranged between the cylindrical body and the pot core; the outer shell of the centrifugal pot is fixedly connected to the cylindrical body and the filtrate space is closed by the cylindrical body.

[0007] Furthermore, an inlet is provided at the upper end of the pot core, and a rolling ring is fixedly connected to the outer side of the inlet; a universal ball is installed on the outer shell of the centrifugal pot corresponding to the rolling ring, and the rolling ring is in contact with the universal ball.

[0008] Furthermore, the volumes of the primary centrifugal pot, the secondary centrifugal pot and the tertiary centrifugal pot decrease in sequence.

[0009] Furthermore, cooling pipes are arranged in the bases on both sides of the motor.

[0010] The beneficial effects of the utility model are as follows: the utility model provides a centrifugal pot with stepped centrifugation, the pot core arranged inside is provided with an ultrafiltration membrane with successively decreasing pore sizes, and the centrifugal force generated by the rotation of the pot core is used to directly purify and concentrate the protein according to the molecular weight. The ultrafiltration membrane separates the two separated products, the filtrate and the concentrate, so that it is convenient to collect the filtrate and the concentrate separately in the subsequent process without worrying about the problem of secondary mixing, thereby reducing the difficulty of operation, ensuring the purity of protein purification, and improving the efficiency of purification. Secondly, as the centrifugation is carried out in different centrifugal pots, a protein concentrate with a molecular weight cutoff can be obtained in each centrifugal pot, so that it is convenient to classify and utilize proteins of different sizes and activities. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 It is a schematic diagram of the overall structure of the utility model.

[0012] Figure 2 This is a schematic diagram of the structure of the pot core.

[0013] Figure 3 This is an enlarged view of the base structure.

[0014] In the figure:

[0015] 1. Primary centrifugal pot; 2. Secondary centrifugal pot; 3. Tertiary centrifugal pot; 4. Pot core; 5. Filtrate space; 6. External frame; 7. Ultrafiltration membrane; 8. Filtrate outlet pipe; 9. Concentrate outlet pipe; 10. Motor; 11. Base; 12. Lower connecting block; 13. Cylindrical body; 14. Lubricating pad; 15. Outer shell; 16. Inlet; 17. Rolling ring; 18. Universal ball; 19. Cooling pipe; 20. Feed pipe. DETAILED DESCRIPTION

[0016] The following will be combined with the drawings in the embodiments of the utility model to clearly and completely describe the technical solutions in the embodiments of the utility model. Obviously, the described embodiments are only part of the embodiments of the utility model, not all of the embodiments. Based on the embodiments in the utility model, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the utility model.

[0017] like Figure 1The extraction and purification device of an active protein shown in the figure includes a primary centrifugal pot 1, a secondary centrifugal pot 2 and a tertiary centrifugal pot 3; the three centrifugal pots can be used in combination to perform step-by-step separation and purification of proteins. The volumes of the primary centrifugal pot 1, the secondary centrifugal pot 2 and the tertiary centrifugal pot 3 decrease successively as the capacity of the processed protein liquid decreases. And the filter pore size decreases successively to concentrate small molecular proteins. For example, the ultrafiltration membrane 7 in the primary centrifugal pot 1 can pass molecular weights less than 50kd, the ultrafiltration membrane 7 in the secondary centrifugal pot 2 can pass molecular weights less than 30kd, and the tertiary centrifugal pot 3 can pass molecular weights less than 10kd or 5kd. Thus, the small molecular proteins are separated and concentrated from the large molecular proteins step by step.

[0018] Specifically, each centrifugal pot is rotatably connected to a pot core 4, and a gap is left between the pot core 4 and the centrifugal pot shell 15 to form a filtrate space 5; Figure 2 As shown, the pot core 4 includes an exoskeleton 6 made of steel in the shape of a icosahedron, an inner frame of the same shape as the exoskeleton 6, and an ultrafiltration membrane 7 clamped between the exoskeleton 6 and the inner frame. Thus, when the pot core 4 rotates, the protein molecules that can pass through the filter membrane are centrifuged into the filtrate space 5, while the large molecular weight proteins that cannot pass through the ultrafiltration membrane 7 are retained in the pot core 4. The icosahedron-shaped pot core 4 can prevent the internal liquid level from rising and escaping to the outside of the pot core 4 during rotation, so that the pot core 4 can be centrifuged without leakage even without a cover.

[0019] The pore size of the ultrafiltration membrane 7 in the primary centrifugal pot 1 is the largest, and the pore size of the ultrafiltration membrane 7 in the tertiary centrifugal pot 3 is the smallest, thereby realizing step-by-step separation. A filtrate outlet pipe 8 is installed on the centrifugal pot shell 15, and the filtrate outlet pipe 8 is connected to the filtrate space 5, and the other end of the filtrate outlet pipe 8 is connected to the pot core 4 of the next-stage centrifugal pot. After the upper-stage centrifugal pot stops working, the filtered filtrate can be discharged into the pot core 4 of the next-stage centrifugal pot through the filtrate outlet pipe 8, so as to facilitate the next level of centrifugal concentration. A concentration outlet pipe 9 connected to the outside world is provided in the pot core 4; the concentration outlet pipe 9 is suspended in the pot core 4 and is not affected by the rotation of the pot core 4. After the centrifugation is completed, the concentrated and retained protein liquid can be discharged through the concentration outlet pipe 9 for timely collection. A motor 10 is provided at the bottom of the centrifugal pot, and the motor 10 is transmission-connected to the bottom of the pot core 4.

[0020] In this embodiment, Figure 3 As shown, the bottom of the centrifugal kettle is mounted on the base 11, and the motor 10 is mounted in the base 11. A lower connecting block 12 is fixed to the outer frame 6 at the lower end of the kettle core 4; the output shaft of the motor 10 is in driving connection with the lower connecting block 12. The motor 10 provides driving force to rotate the kettle core 4. In order to prevent the ultrafiltration membrane 7 from being damaged by pressure, a low-speed and long-term centrifugation method is used, so the requirements for the motor 10 are reduced, and the requirements for the stability of the rotation of the kettle core 4 are also reduced.

[0021] A cylindrical body 13 is provided at the upper end of the base 11, the lower connecting block 12 is located in the cylindrical body 13, and a lubricating pad 14 is provided between the cylindrical body 13 and the pot core 4; the centrifugal pot shell 15 is fixedly connected to the cylindrical body 13 and closes the filtrate space 5 through the cylindrical body 13.

[0022] The upper end of the pot core 4 is provided with an inlet 16, and a rolling ring 17 is fixedly connected to the outer side of the inlet 16; a universal ball 18 is installed on the centrifugal pot shell 15 corresponding to the rolling ring 17, and the rolling ring 17 contacts the universal ball 18. The rolling ring 17 changes the outer shape of the inlet 16 into a circle to facilitate the rotation of the pot core 4. The universal ball 18 can provide the upper part of the pot core 4 with a rotation support force to prevent the pot core 4 from tilting during rotation.

[0023] In this embodiment, cooling pipes 19 are provided in the bases 11 on both sides of the motor 10 to cool the motor 10 and prevent the heat of the motor 10 from being transferred to the protein solution and causing protein denaturation.

[0024] When in use, the initial protein solution enters the pot core 4 from the feed pipe 20 of the primary centrifugal pot 1, and then centrifuges to separate, purify and concentrate the initial protein solution according to the molecular weight. The remaining protein solution after centrifugation to remove the macromolecular protein is sent to the secondary centrifugal pot 2 through the filtrate outlet pipe 8 of the primary centrifugal pot 1 for secondary centrifugation. The concentrated solution after centrifugation in the pot core 4 of the primary centrifugal pot 1 is discharged and collected through the concentrated outlet pipe 9. After the secondary centrifugal pot 2 is finished working, the filtrate produced is sent to the pot core 4 of the tertiary centrifugal pot 3 from the filtrate outlet pipe 8 of the secondary centrifugal pot 2. The protein solution retained in the secondary centrifugal pot 2 is discharged and collected from the concentrated outlet pipe 9 of the secondary centrifugal pot 2. The protein filtrate produced after centrifugation in the tertiary centrifugal pot 3 contains only small molecular weight proteins (not exceeding 10kd or 5kd), and is discharged from the filtrate outlet pipe 8 of the tertiary centrifugal pot 3. The protein solution retained in the pot core 4 of the tertiary centrifugal pot 3 is discharged and collected from the concentrated outlet pipe 9 of the tertiary centrifugal pot 3. In this way, the initial protein solution can be separated and purified according to the molecular weight, and a protein solution with any molecular retention amount can be collected.

[0025] It is obvious to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the present invention can be implemented in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments should be regarded as exemplary and non-restrictive from any point of view, and the scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims be included in the present invention. Any reference numeral in a claim should not be regarded as limiting the claim to which it relates.

Claims

1. An extraction and purification device for active protein, characterized in that: It comprises a primary centrifugal pot, a secondary centrifugal pot and a tertiary centrifugal pot; each centrifugal pot is rotatably connected to a pot core inside, and a gap is left between the pot core and the outer shell of the centrifugal pot to form a filtrate space; the pot core comprises an outer skeleton made of steel in the shape of a icosahedron, an inner skeleton with the same shape as the outer skeleton, and an ultrafiltration membrane clamped between the outer skeleton and the inner skeleton; the pore size of the ultrafiltration membrane in the primary centrifugal pot is the largest, and the pore size of the ultrafiltration membrane in the tertiary centrifugal pot is the smallest; a filtrate outlet pipe is installed on the outer shell of the centrifugal pot, the filtrate outlet pipe is connected to the filtrate space, and the other end of the filtrate outlet pipe is connected to the pot core of the next-stage centrifugal pot; a concentration outlet pipe connected to the outside is arranged in the pot core; a motor is arranged at the bottom of the centrifugal pot, and the motor is transmission-connected to the bottom of the pot core.

2. The active protein extraction and purification device according to claim 1, characterized in that: The bottom of the centrifugal pot is installed on the base, and the motor is installed in the base; a lower connecting block is fixed on the outer frame at the lower end of the pot core; and the output shaft of the motor is drivingly connected with the lower connecting block.

3. The active protein extraction and purification device according to claim 2, characterized in that: A cylindrical body is arranged at the upper end of the base, the lower connecting block is located in the cylindrical body, and a lubricating pad is arranged between the cylindrical body and the pot core; the outer shell of the centrifugal pot is fixedly connected to the cylindrical body and closes the filtrate space through the cylindrical body.

4. The active protein extraction and purification device according to claim 3, characterized in that: An inlet is arranged at the upper end of the pot core, and a rolling ring is fixedly connected to the outer side of the inlet; a universal ball is installed on the outer shell of the centrifugal pot corresponding to the rolling ring, and the rolling ring is in contact with the universal ball.

5. The active protein extraction and purification device according to claim 4, characterized in that: The volumes of the primary centrifugal pot, the secondary centrifugal pot and the tertiary centrifugal pot decrease in sequence.

6. The active protein extraction and purification device according to claim 5, characterized in that: Cooling pipes are arranged in the bases on both sides of the motor.

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