Low-temperature protein purification equipment for nano antibody

Abstract

The low-temperature protein purification equipment comprises an outer barrel, a sealing cover is arranged on the top wall of the outer barrel, a fixing frame is arranged on the top wall of the sealing cover, a rotating groove is formed in the bottom wall of the fixing frame, and a rotating ring is arranged on the inner wall of the rotating groove. According to the technical scheme, through cooperation of the outer barrel, the fixing frame, the low-temperature pipe and other devices, the low-temperature pipe can be conveniently driven by the devices to rotate, the inner cavity of the outer barrel can be conveniently and evenly cooled, the inner cavity of the outer barrel can be evenly cooled, and the cooling effect of the outer barrel is improved; the nanometer antibody protein can be conveniently purified and prepared in a low-temperature state; through cooperation of the inner barrel, the inner membrane, the outer barrel and the like, the device is convenient for large-scale purification of nano antibody protein, and has the advantages of high liquid outflow speed and high purification efficiency.

Description

TECHNICAL FIELD

[0001] The utility model belongs to the technical field of protein purification equipment, specifically to a kind of low temperature protein purification equipment for nanobody. BACKGROUND

[0002] Biopharmaceutical is a pharmaceutical industry field that arises and develops vigorously in the 21st century, and the main categories include recombinant protein drugs, monoclonal antibody drugs, vaccines and the like, which play an irreplaceable role in the treatment or prevention of various major diseases such as malignant tumors, autoimmune diseases, genetic diseases and infectious diseases, and protein purification is an important step in the production process of biological products, and the preparation of part of nanobody protein is in a low temperature environment, and the application number: 202222771530.9 discloses a protein purification equipment, which belongs to a biological pharmaceutical production equipment and is used for large-scale purification of recombinant proteins, especially monoclonal antibodies, compared with a traditional liquid chromatography purification column, the equipment is more suitable for large-scale purification of recombinant proteins, especially antibody drugs, has the advantages of fast liquid outflow speed and high purification efficiency, and the protein purification equipment is not convenient for protein cooling and purification, it is difficult to uniformly cool, and it is not convenient to prepare low-temperature proteins, so as to avoid the influence of inconsistent temperature on protein purification and preparation. SUMMARY

[0003] In view of the above problems, the low temperature protein purification equipment for nanobody is provided to effectively solve the problems that the protein purification equipment is not convenient for protein cooling and purification, it is difficult to uniformly cool, and it is not convenient to prepare low-temperature proteins, so as to avoid the influence of inconsistent temperature on protein purification and preparation.

[0004] To achieve the above purpose, the utility model provides the following technical scheme: a kind of low temperature protein purification equipment for nanobody, including outer bucket, the top wall of the outer bucket is provided with closure cover, the top wall of the closure cover is provided with fixed frame, the bottom wall of the fixed frame is opened with rotation groove, the inner wall of the rotation groove is provided with swivel ring, the bottom wall of the swivel ring is provided with low temperature pipe, the outer wall and inner wall of the rotation groove are respectively provided with liquid inlet groove and liquid outlet groove, the left and right sides of the outer wall of the fixed frame are respectively provided with liquid inlet pipe and liquid outlet pipe, the liquid inlet pipe is communicated with the liquid inlet groove, the liquid outlet pipe is communicated with the liquid outlet groove, the inner wall of the liquid inlet groove and the liquid outlet groove is provided with liquid guide ring, the outer wall of the liquid guide ring is opened with through-hole, the left and right ends of the low temperature pipe are respectively arranged on the inner wall of the through-hole on the left and right sides, the top wall of the fixed frame is provided with motor, the outer wall of the low temperature pipe is provided with transmission frame, and the output end of the motor is connected with the top end of the transmission frame.

[0005] Preferably, the inner wall of the outer barrel is provided with an inner barrel, the inner wall of the inner barrel is uniformly provided with holes, the inner wall of the inner barrel is provided with an inner film, the bottom wall of the outer barrel is provided with a discharge port, and the top wall of the sealing cover is provided with a feeding port.

[0006] Preferably, the outer wall of the transmission frame is uniformly provided with stirring fins, and the stirring fins are arranged on the outer wall of the low-temperature pipe.

[0007] Preferably, the outer barrel and the inner barrel are made of stainless steel, and the diameter of the holes on the inner barrel wall ranges from 0.1 to 10 mm.

[0008] Preferably, the inner wall of the inner barrel is provided with a limiting ring, the outer wall of the limiting ring is uniformly provided with buckles, the inner film is a perforated film, and the hole diameter is uniform.

[0009] Compared with the prior art, the device has the advantages that:

[0010] 1. The nano antibody low-temperature protein purification device can drive the low-temperature pipe to rotate, uniformly cool the inner cavity of the outer barrel, and prepare nano antibody protein under low-temperature conditions.

[0011] 2. The nano antibody low-temperature protein purification device can purify a large amount of nano antibody protein, has the advantages of fast liquid outflow speed and high purification efficiency.

[0012] 3. The nano antibody low-temperature protein purification device can stir and cool the raw materials, uniformly cool and stir, and perform purification work under a low-temperature environment. BRIEF DESCRIPTION OF DRAWINGS

[0013] The accompanying drawings are used to provide a further understanding of the present application, and constitute a part of the specification, together with the embodiments of the present application, to explain the present application, and do not constitute a limitation on the present application.

[0014] In the drawings:

[0015] Figure 1 It is a structure schematic view of the nano antibody low-temperature protein purification device of the present application.

[0016] Figure 2 It is a sectional structure schematic view of the outer barrel of the present application.

[0017] Figure 3 It is a sectional structure schematic view of the outer barrel of the present application. Figure 2 A enlarged view of the A part of the present application

[0018] In the figure: 100, outer barrel; 101, sealing cover; 102, fixed frame; 103, swivel ring; 104, low temperature pipe; 105, liquid inlet pipe; 106, liquid outlet pipe; 107, liquid guide ring; 108, motor; 109, transmission frame; 110, inner barrel; 111, inner membrane; 112, discharge port; 113, feed port; 114, stirring fin; 115, limiting ring; 116, buckle. DETAILED DESCRIPTION

[0019] The technical solutions in the embodiments of the utility model will be clearly and completely described below with reference to the drawings in the embodiments of the utility model. Obviously, the described embodiments are only part of the embodiments of the utility model, rather than all the embodiments. Based on the embodiments in the utility model, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the utility model.

[0020] Given by Figure 1 , Figure 2 and Figure 3 The utility model discloses a kind of low temperature protein purification equipment for nanobody, the top wall of outer barrel 100 is equipped with sealing cover 101, outer barrel 100 is sealed by sealing cover 101, the top wall of sealing cover 101 is provided with fixed frame 102, swivel ring 103 is supported by fixed frame 102, the bottom wall of fixed frame 102 is equipped with rotation groove, the inner wall of rotation groove is provided with swivel ring 103, swivel ring 103 is closed to the inner wall of rotation groove by, the bottom wall of swivel ring 103 is fixedly connected with low temperature pipe 104, the inner cavity of outer barrel 100 is cooled by low temperature pipe 104, the outer wall and the inner wall of rotation groove are equipped with liquid inlet groove and liquid outlet groove respectively, the left and right sides of fixed frame 102 outer wall are fixedly connected with liquid inlet pipe 105 and liquid outlet pipe 106 respectively, liquid inlet pipe 105 is communicated with liquid inlet groove, liquid outlet pipe 106 is communicated with liquid outlet groove, the inner wall of liquid inlet groove and liquid outlet groove is rotatably connected with liquid guide ring 107, liquid inlet groove and liquid outlet groove are closed to the inner wall by liquid guide ring 107, the outer wall of liquid guide ring 107 is equipped with through-hole, the left and right ends of low temperature pipe 104 are respectively arranged in the inner wall of left and right sides through-hole, the top wall of fixed frame 102 is fixedly connected with motor 108, transmission frame 109 is rotatable by motor 108, the outer wall of low temperature pipe 104 is fixedly connected with transmission frame 109, low temperature pipe 104 is rotated by transmission frame 109, the output end of motor 108 is connected with the top end of transmission frame 109.

[0021] In this embodiment: the prepared raw materials are injected into the inner cavity of the outer barrel 100, and then the closure cover 101 is closed to seal the outer barrel 100. The liquid inlet pipe 105 and the liquid outlet pipe 106 are connected to the cooling machine through the pipeline. The low-temperature cooling liquid is injected into the liquid inlet pipe 105 and the liquid outlet pipe 106 through the cooling machine. The liquid inlet pipe 105 injects into the liquid inlet groove, and the cooling liquid is injected into the inner cavity of the low-temperature pipe 104 through the liquid inlet groove, so that the cooling liquid enters the low-temperature pipe 104 for cooling and cooling. Then the cooling liquid flows out from the right end of the low-temperature pipe 104 into the liquid outlet groove, and the cooling liquid is injected into the inner cavity of the liquid outlet pipe 106 for discharge, and then flows back to the cooling machine through the conduit. The motor 108 drives the transmission frame 109 to rotate, and the low-temperature pipe 104 is driven by the transmission frame 109 to rotate. The rotation of the low-temperature pipe 104 facilitates uniform cooling of the raw materials. The device can conveniently drive the low-temperature pipe 104 to rotate, facilitate uniform cooling of the inner cavity of the outer barrel 100, and facilitate nanobody protein purification under low-temperature conditions.

[0022] The inner wall of the outer barrel 100 is provided with an inner barrel 110. The inner wall of the inner barrel 110 is uniformly provided with holes. The inner wall of the inner barrel 110 is fixedly connected with an inner membrane 111. The bottom wall of the outer barrel 100 is fixedly connected with a discharge port 112. The top wall of the closure cover 101 is provided with a feed port 113.

[0023] In this embodiment: first step: select the appropriate pore size of the inner membrane 111 according to the particle size of the purification medium gel beads (such as agarose gel, dextran gel, etc.). Second step: close the discharge port 112 and open the feed port 113. The liquid containing the purification medium is poured into the inner membrane 111 through the feed port 113 and is placed for 10-30 minutes. Third step: the solution containing the protein to be purified is poured into the inner membrane 111 through the feed port 113. Fourth step: start the motor 108 to drive the low-temperature pipe 104 to rotate for cooling and stirring. After uniform stirring, place for 10-2 hours. Then open the discharge port 112 to discharge the liquid in the container. The gel beads are trapped in the inner membrane 111. Fifth step: then sequentially pour the washing liquid, eluent and other liquids into the inner membrane 111 through the feed port 113. After each addition, place for 10-30 minutes and repeat the fourth step until the protein to be purified is eluted. The bottom wall of the outer barrel 100 can also be installed with a differential machine to drive the inner barrel 110 to rotate for liquid spinning, so as to utilize the centrifugal force for rapid liquid discharge. The device can facilitate large-scale purification of nanobody protein, has the advantages of fast liquid outflow speed and high purification efficiency.

[0024] The outer wall of the transmission frame 109 is uniformly fixedly connected with stirring fins 114. The stirring fins 114 are made of a material that facilitates temperature conduction. The stirring fins 114 are arranged on the outer wall of the low-temperature pipe 104.

[0025] In the embodiment: by setting transmission frame 109 facilitate the stirring fin 114 rotation, through the stirring fin 114 rotation facilitates the raw materials for stirring, while through the stirring fin 114 facilitates the conduction of temperature, can make the device is convenient for raw materials for stirring cooling, convenient for uniform cooling stirring, facilitate the purification operation in low temperature environment.

[0026] The outer barrel 100 and the inner barrel 110 are made of stainless steel material, the diameter of the hole on the wall of the inner barrel 110 is 0.1-10mm.

[0027] In the embodiment: the use of stainless steel material can avoid rust, improve the practicability of the device, facilitate the subsequent protein purification operation.

[0028] The inner wall of the inner barrel 110 is fixedly connected with a limiting ring 115, which supports the inner membrane 111. The outer wall of the limiting ring 115 is uniformly connected with a buckle 116. The inner membrane 111 is a perforated membrane with uniform hole diameter.

[0029] In the embodiment: through the limiting ring 115 and the buckle 116 cooperate with the inner membrane 111 for limiting and fixing, convenient for installation and fixing, convenient for subsequent purification operation.

Claims

1. A low-temperature protein purification device for nanobodies, comprising an outer container (100), characterized in that: The top wall of the outer barrel (100) is provided with a sealing cover (101), and the top wall of the sealing cover (101) is provided with a fixing frame (102). The bottom wall of the fixing frame (102) is provided with a rotating groove, and the inner wall of the rotating groove is provided with a rotating ring (103). The bottom wall of the rotating ring (103) is provided with a low-temperature tube (104). The outer wall and inner wall of the rotating groove are respectively provided with a liquid inlet groove and a liquid outlet groove. The left and right sides of the outer wall of the fixing frame (102) are respectively provided with a liquid inlet pipe (105) and a liquid outlet pipe (106). The liquid inlet pipe (105) and the... The inlet tank is connected to the outlet tank, and the outlet pipe (106) is connected to the outlet tank. The inner walls of the inlet tank and the outlet tank are provided with liquid guide rings (107). The outer walls of the liquid guide rings (107) are provided with through holes. The left and right ends of the low temperature tube (104) are respectively provided on the inner walls of the through holes on the left and right sides. The top wall of the fixing frame (102) is provided with a motor (108), and the outer wall of the low temperature tube (104) is provided with a transmission frame (109). The output end of the motor (108) is connected to the top end of the transmission frame (109).

2. The low-temperature protein purification device for nanobodies according to claim 1, characterized in that: The inner wall of the outer barrel (100) is provided with an inner barrel (110), the inner wall of the inner barrel (110) is provided with holes evenly distributed, the inner wall of the inner barrel (110) is provided with an inner membrane (111), the bottom wall of the outer barrel (100) is provided with a discharge port (112), and the top wall of the sealing cover (101) is provided with a feed port (113).

3. The low-temperature protein purification device for nanobodies according to claim 1, characterized in that: The outer wall of the transmission frame (109) is uniformly provided with stirring fins (114), which are disposed on the outer wall of the low temperature tube (104).

4. The low-temperature protein purification device for nanobodies according to claim 2, characterized in that: The outer barrel (100) and the inner barrel (110) are made of stainless steel, and the diameter of the hole on the wall of the inner barrel (110) ranges from 0.1 to 10 mm.

5. The low-temperature protein purification device for nanobodies according to claim 2, characterized in that: The inner wall of the inner barrel (110) is provided with a limiting ring (115), and the outer wall of the limiting ring (115) is uniformly provided with buckles (116). The inner membrane (111) is a perforated membrane with uniform pore size.

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