A chromatography column packing system and a chromatography column packing method

Abstract

The present application relates to a kind of chromatographic column packing system and chromatographic column packing method, belong to the technical field of separation and purification, the end cap assembly center of chromatographic column packing system is provided with gas / liquid mixed valve, the inner surface of end cap assembly is provided with lower sieve plate and lower distribution plate;The center of piston assembly is provided with multi-way valve, the inner surface of piston assembly is provided with upper sieve plate, multi-way valve is provided with at least sieve plate exhaust valve, piston assembly internal exhaust valve, mobile phase valve.Chromatographic column packing method is in the column barrel inside and is filled with packing, standing, exhaust, compressed column bed, completes the process of packing column.The present application can improve the packing efficiency of large chromatographic column, so that packing reaches ideal height while having good column efficiency, and the method is simple to implement, efficient and stable.

Description

Technical Field

[0001] This invention relates to a chromatography column packing system and a chromatography column packing method, belonging to the technical field of separation and purification. Background Technology

[0002] Chromatography can be used to effectively separate target products from complex mixtures, remove impurities from target substances, purify final products, or replace product solutions. It features high separation efficiency and can separate heat-sensitive or easily inactivated products, such as proteins, polysaccharides, and peptides, under relatively mild conditions. The chromatography column is the key to achieving chromatographic separation.

[0003] Chromatography columns are essential equipment in the chromatography industry for purifying process liquids and separating desired substances from them. They are typically used in large-scale preparative purification of biological products by major extract manufacturers, pharmaceutical companies, cosmetics companies, and food companies. With industrial development, automated chromatography columns, due to their high degree of automation, process controllability, and high chromatography efficiency, have gradually become the mainstream equipment in the chromatography industry.

[0004] The current mainstream method for packing chromatography columns is as follows: before loading the packing material into the column, air bubbles inside the column (including the upper and lower sieve plates and the inside of the tubing) are removed. The packing material is first homogenized in a homogenizer, and then the homogenized packing material is loaded into the column. If the packing effect is not good, the packing material in the column needs to be pushed from the inside of the column into the homogenizer, and the packing material needs to be returned to the homogenizer for homogenization again. The column needs to be cleaned and vented again, which is very time-consuming and can also damage the packing material, especially for large-diameter chromatography columns.

[0005] To address the shortcomings of existing technologies, this application provides a chromatography column packing system and a chromatography column packing method that eliminates the need to return poorly packed packing material to the homogenizer and avoids operations such as re-cleaning the column. Summary of the Invention

[0006] To address the aforementioned problems, this invention discloses a chromatography column packing system and a chromatography column packing method, the specific technical solution of which is as follows: A chromatography column packing system includes a column cylinder, an end cap assembly at the bottom of the column cylinder, and a piston assembly at the top. A gas / liquid mixing valve is provided at the center of the end cap assembly, and a lower distribution plate is provided on the inner surface of the end cap assembly. A multi-way valve is provided at the center of the piston assembly, and an upper distributor is provided on the lower surface of the piston assembly. The upper distributor has a channel at its center for the multi-way valve to pass through. The lower surface of the upper distributor rises almost from the periphery toward the center, forming a conical surface that extends upward from the periphery to the center. The lower surface of the upper distributor is covered with an upper sieve plate, which is also a conical surface extending upward from the periphery to the center, consistent with the shape of the lower surface of the upper distributor.

[0007] Furthermore, the upper surface of the end cap assembly is provided with a lower distributor. The upper surface of the lower distributor is a conical surface that extends downward from the periphery to the center, and a lower sieve plate with the same shape direction is laid on it. The lower distributor plate is located between the lower sieve plate and the lower distributor.

[0008] Furthermore, the lower distribution plate has a central hole at its center, with the front of the lower distribution plate facing downwards. The front of the lower distribution plate has several elongated, downward-protruding distribution teeth that radiate outwards from the center hole toward the edge of the lower distribution plate. Between adjacent distribution teeth, there are half-distribution teeth that radiate outwards from the periphery of the lower distribution plate to the angle between adjacent distribution teeth. The lower distribution plate has several through holes between adjacent distribution teeth and between distribution teeth and their adjacent half-distribution teeth.

[0009] Furthermore, the multi-way valve is equipped with at least an upper screen plate exhaust valve, a piston assembly internal exhaust valve, and a flow phase valve; the flow phase inlet / outlet pipe connected to the flow phase valve passes through the center of the piston assembly and extends beyond the upper screen plate at the bottom; the exhaust channel connected to the upper screen plate exhaust valve has its bottom flush with the center of the upper screen plate and surrounds the flow phase inlet / outlet pipe; the exhaust channel connected to the piston assembly internal exhaust valve is located at the upper edge inside the piston assembly.

[0010] Furthermore, the upper sieve plate has a multi-layer mesh structure.

[0011] A chromatography column packing method based on the above-mentioned chromatography column packing system involves dispersing, settling, venting, and compressing the column bed inside the column to complete the packing process. The specific process is as follows: Step 1: Packing material dispersion: The packing material is homogenized in a homogenizing tank to obtain uniformly dispersed packing material; Step 2: Column packing: Pack the packing material that was broken up in Step 1 into the chromatography column. If the packing material settles naturally and forms a uniform column, the column packing is complete and a qualified column bed is formed. If there are air bubbles in the packing material or the column efficiency is not up to standard, proceed to Step 3. Step 3: Inject buffer solution: Raise the piston assembly and inject buffer solution into the column at the same time during the raising process. After the column is filled with buffer solution, use compressed air to blow the column bed and the upper sieve plate to completely break up the column bed. The surface of the upper sieve plate needs to be kept dry. Step 4: Column bed sedimentation: After the packing material is evenly dispersed, it settles naturally, or the buffer solution is pumped into the upper end of the column and the buffer solution is pumped out from the lower end of the column to compact the column bed. Step 5: Column venting: First, close the gas / liquid mixing valve of the end cap assembly. The inside of the column is connected to the outside through the multi-port valve of the piston assembly. Press down the piston assembly, and the buffer solution will be discharged from the multi-port valve, bringing out the air at the same time. Then close the multi-port valve of the piston assembly and open the gas / liquid mixing valve of the end cap assembly. At this time, the inside of the column can only be connected to the outside through the gas / liquid mixing valve of the end cap assembly. Continue to press down the piston assembly, and the buffer solution will be discharged from the gas / liquid mixing valve of the end cap assembly, squeezing out the air bubbles inside the gas / liquid mixing valve of the end cap assembly. Step 6: Compress the packing: The lowering piston assembly compresses the packing, stopping when it reaches the target position according to the compressibility factor, thus completing the packing.

[0012] Furthermore, in step 2, the packing is pumped into the column through the multi-port valve of the piston assembly, or drawn into the column through the gas / liquid mixing valve of the end cap assembly.

[0013] Furthermore, in step 3, the upper screen plate is kept dry before venting. The method to keep the surface of the upper screen plate dry is to remove the upper screen plate from the liquid surface of the column before venting, and then blow the upper screen plate dry with compressed gas.

[0014] Furthermore, in step 3, compressed air is blown into the packing column from the gas / liquid mixing valve of the end cap assembly, and the blowing continues until the packing is uniform, with the compressed air pressure not less than 1.5 Bar.

[0015] Furthermore, in step 5, the air bubbles on the surface of the sieve plate are vented by pressing down the piston assembly, or by pumping buffer solution into the column while the piston assembly is stationary.

[0016] The beneficial effects of this invention are: This invention solves the problems of long time consumption and high losses in conventional methods (external packing). It enables the mixing of uneven packing inside the column by blowing, and at the same time, it can remove air bubbles, which greatly improves the efficiency of column packing and saves time and procedures.

[0017] The conical design of the upper sieve plate in this invention allows bubbles to move in a concentrated manner to the center of the piston assembly, facilitating the smooth removal of bubbles.

[0018] The lower distribution plate of the present invention enables the airflow to be evenly diffused to the lower sieve plate, which is beneficial to dispersing the packing material inside the column. Attached Figure Description

[0019] Figure 1 This is a schematic diagram of the method process of the present invention. Figure 2 This is a longitudinal sectional view of the chromatography column packing system of the present invention. Figure 3 yes Figure 2A schematic diagram showing the state of the packing being dispersed by the air intake of a multi-way valve. Figure 4 These are schematic diagrams of the front and back sides of the lower distribution plate of the present invention. Figure 5 This is a schematic diagram of the upper distributor of the present invention.

[0020] List of reference numerals in the attached diagram: 1—Piston assembly, 2—Column, 3—Lower sieve plate, 4—Lower distribution plate, 5—Lower distributor, 6—End cap assembly, 7—Gas / liquid mixing valve, 8—Multi-port valve, 9—Upper distributor, 10—Through hole, 11—Upper sieve plate, 12—Distribution teeth, 13—Half-section distribution teeth, 14—Vent hole, a—Upper sieve plate exhaust valve, a1—Exhaust passage connected to the upper sieve plate exhaust valve, b—Flow phase valve, b1—Flow phase inlet / outlet pipe connected to the flow phase valve, c—Internal exhaust valve of piston assembly, c1—Exhaust passage connected to the internal exhaust valve of piston assembly. Detailed Implementation

[0021] The present invention will be further illustrated below with reference to the accompanying drawings and specific embodiments. It should be understood that the following specific embodiments are for illustrative purposes only and are not intended to limit the scope of the invention.

[0022] Appendix Figure 2 , 3 The magenta lines in section 5 are auxiliary lines and are used for reference only. Figure 2 , 3 The middle serves as a flow path for gases or liquids. Figure 5 The center serves as a reference for the horizontal plane.

[0023] like Figure 1 As shown, this invention provides a column packing method for chromatography columns, addressing situations where column efficiency fails to meet requirements after packing material is introduced, i.e., the column efficiency is unsatisfactory after packing material is introduced. In this case, the invention eliminates the need to return the packing material for re-homogenization; instead, re-homogenization and air bubble removal are performed directly within the column cylinder 2. Specifically, the packing material is dispersed, allowed to settle, vented, and the column bed compressed inside the column cylinder 2.

[0024] like Figure 1 As shown, when the column efficiency is not up to standard, the buffer solution should first be pumped or drawn into the column 2. During the process of pumping or drawing in the buffer solution, the piston assembly 1 is raised upward to twice the height of the column bed to provide space for the buffer solution.

[0025] After the buffer injection is complete, see Figure 3 Compressed air is blown in from the bottom of column 2 at a pressure of 1.5 kg to blow and mix the buffer solution and packing material, thus breaking up the packing material and forming a homogenate.

[0026] After allowing the homogenized filler to settle naturally for a period of time, proceed with the operation of the lower piston assembly 1, as described above. Figure 2 The buffer solution is discharged from the multi-port valve 8 of the piston assembly 1, and the air bubbles in the column 2 are also concentrated at the top of the buffer solution and discharged along with it. When the piston assembly 1 approaches or contacts the packing material, the multi-port valve 8 of the piston assembly 1 is closed, and the gas / liquid mixing valve 7 of the end cap assembly 6 is opened. At this time, the inside of the column 2 can only communicate with the outside through the gas / liquid mixing valve 7 of the end cap assembly 6. Continuing to press down the piston assembly 1, the buffer solution is discharged from the gas / liquid mixing valve 7 of the end cap assembly 6, squeezing out the air bubbles inside the gas / liquid mixing valve 7 of the end cap assembly 6. In this way, all the air bubbles inside the column 2 can be completely removed.

[0027] Finally, compact the packing material, compressing it to the target position according to the compressibility factor, and stop to complete the column loading.

[0028] The following is combined with Figure 2-5 The structural design of the chromatography column packing system described in this invention is explained in detail below: The bottom of the column 2 is equipped with an end cap assembly 6, and a gas / liquid mixing valve 7 is located in the center of the end cap assembly 6. The main functions of the end cap assembly 6 are: 1. To pressurize the buffer solution into the column 2 through the gas / liquid mixing valve 7. Pressurizing the buffer solution from the bottom can disperse the packing material due to the force of the buffer solution entering the column 2. The buffer solution can also be injected from the piston assembly 1 at the top of the column 2, or simultaneously from the bottom and top of the column 2, or injected in a certain proportion, or a certain amount can be injected from the top of the column 2 first to allow the packing material to flow before injecting from the bottom. 2. To blow compressed gas into the column 2 through the gas / liquid mixing valve 7, allowing the high-pressure gas to disperse the packing material and achieve homogenization of the packing material inside the column 2. 3. During the stage of purging buffer and air bubbles, after the buffer and air bubbles in the column 2 are mainly discharged from the piston assembly 1 at the top of the column 2, close the multi-port valve 8 of the piston assembly 1, open the gas / liquid mixing valve 7 at the bottom of the column 2, squeeze out the buffer from the gas / liquid mixing valve 7, and carry out the air bubbles in the gas / liquid mixing valve 7 channel, so as to completely remove the air bubbles inside the column 2.

[0029] A lower distributor 5 is provided on the upper surface of the end cap assembly 6. The upper surface of the lower distributor 5 is a tapered surface extending downwards from the periphery to the center, and a lower sieve plate 3 with the same shape orientation is laid on it. A lower distribution plate 4 is located between the lower sieve plate 3 and the lower distributor 5. See also Figure 4The lower distribution plate 4 has a central hole at its center, with its front facing downwards. Several elongated, downward-protruding distribution teeth 12 radiate from the central hole towards the edge of the lower distribution plate 4. Between adjacent distribution teeth 12, there are semi-distribution teeth 13 extending from the periphery of the lower distribution plate 4 to the angle between adjacent distribution teeth 12. Several vent holes 14 are provided between adjacent distribution teeth 12 and between distribution teeth 12 and their adjacent semi-distribution teeth 13. The lower distributor 5, lower screen plate 3, and lower distribution plate 4 work together to regulate the airflow of compressed gas entering the column cylinder 2. The downward-protruding teeth 12 disperse the airflow by blowing air upwards. Combined with the multiple vent holes 14, this ensures both uniform distribution of the mobile phase during equipment use and provides uniform airflow to disperse the packing material during aeration and homogenization.

[0030] A multi-way valve 8 is provided at the center of the piston assembly 1. After the packing in the column cylinder 2 is evenly dispersed by the buffer solution, the air bubbles and buffer solution in the column cylinder 2 are discharged from the multi-way valve 8 during the downward pressing of the piston assembly 1.

[0031] The piston assembly 1 has an upper distributor 9 on its lower surface, see [reference]. Figure 5 The upper distributor 9 has a through hole 10 at its center for the multi-way valve 8 to pass through. The lower surface of the upper distributor 9 rises almost upwards from the periphery towards the center, forming a conical surface extending upwards from the periphery to the center. An upper sieve plate 11 is laid on the lower surface of the upper distributor 9. The shape of the upper sieve plate 11 is also a conical surface extending upwards from the periphery to the center, consistent with the shape of the lower surface of the upper distributor 9. The conical surface allows air bubbles from the periphery to flow back to the center, facilitating their concentrated discharge and avoiding dead zones around the periphery, thus preventing air bubble residue.

[0032] See Figure 2The multi-port valve 8 is equipped with at least an upper screen plate exhaust valve a, a piston assembly internal exhaust valve c, and a flow phase valve b; the flow phase inlet / outlet pipe b1, which is connected to the flow phase valve, passes through the center of the piston assembly 1 and extends beyond the upper screen plate 11 at the bottom; the exhaust channel connected to the upper screen plate exhaust valve a has its bottom flush with the center of the upper screen plate 11 and surrounds the flow phase inlet / outlet pipe; the exhaust channel c1, which is connected to the piston assembly internal exhaust valve c, is located at the upper edge inside the piston assembly 1. The following describes the use of the multi-way valve 8 in detail: During the stage of draining buffer and bubbles, first open the upper sieve plate exhaust valve a, and lower the piston assembly 1. The air inside the cylinder 2 assembly is discharged from the upper sieve plate exhaust valve a. During the downward pressing of the piston assembly 1, there will be a certain positive pressure inside the piston assembly. The pressure inside the piston assembly 1 can be controlled by controlling the opening and closing of the upper sieve plate exhaust valve a, which is about 1 to 2 kg. Bubbles can only go to the place with low pressure, and the point with low pressure is the outlet of the upper sieve plate exhaust valve a. Continue to lower the piston assembly 1 until the buffer solution is discharged from the upper sieve plate exhaust valve a, and then close the exhaust valve. At this time, the bubbles on the surface of the upper sieve plate 11 have been discharged. The upper sieve plate 11 is set as a conical structure with a high center and low periphery, which makes it easy for air to concentrate in the middle for discharge. Continue to lower the piston assembly 1, and the buffer solution is discharged from the exhaust valve c inside the piston assembly. At the same time, the bubbles inside the upper sieve plate 11 and the bubbles between the upper sieve plate 11 and the piston assembly 1 are discharged. The upper sieve plate 11 is a multi-layer mesh structure. The previous venting only removed the bottom first layer of surface bubbles. After all the air bubbles in piston assembly 1 have been expelled, close the internal exhaust valve c of the piston assembly. Air bubbles near the center of the surface of the upper screen plate 11 will be discharged through the upper screen plate exhaust valve a, while air bubbles at the edges will be discharged through the internal exhaust valve c of the piston assembly. Air bubbles inside the upper screen plate 11 and between the upper screen plate 11 and the upper distributor 9 will be discharged with a slight positive pressure through the port of the internal exhaust valve c of the piston assembly. At this time, the downward pressure inside piston assembly 1 is adjusted to about 1 kg.

[0033] Those skilled in the art will understand that, unless otherwise defined, all terms used herein (including technical and scientific terms) have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. It should also be understood that terms such as those defined in general dictionaries should be understood to have the same meaning as in the context of the prior art, and should not be interpreted in an idealized or overly formal sense unless defined as herein.

[0034] The term "connection" as used in this application can mean a direct connection between components or an indirect connection between components through other components.

[0035] Based on the above-described preferred embodiments of the present invention, and through the foregoing description, those skilled in the art can make various changes and modifications without departing from the inventive concept. The technical scope of this invention is not limited to the contents of the specification, but must be determined according to the scope of the claims.

Claims

1. A chromatography column packing system, comprising a column cylinder (2), characterized in that, The bottom of the cylinder is provided with an end cap assembly (6), and the top is provided with a piston assembly (1). The end cap assembly (6) is provided with a gas / liquid mixing valve (7) at its center, and a lower distribution plate (4) is provided on the inner surface of the end cap assembly (6). The piston assembly (1) is provided with a multi-way valve (8) at its center, and an upper distributor (9) is provided on the lower surface of the piston assembly (1). The upper distributor (9) is provided with a channel for the multi-way valve (8) to pass through at its center. The lower surface of the upper distributor (9) rises from all sides toward the center, forming a conical surface that extends upward from all sides to the center. The lower surface of the upper distributor (9) is covered with an upper sieve plate (11), and the shape of the upper sieve plate (11) is also a conical surface that extends upward from the periphery to the center, which is consistent with the shape direction of the lower surface of the upper distributor (9). The upper surface of the end cap assembly (6) is provided with a lower distributor (5). The upper surface of the lower distributor is a tapered surface that extends downward from the periphery to the center, and a lower sieve plate (3) with the same shape direction is laid on it. The lower distribution plate (4) is located between the lower sieve plate (3) and the lower distributor (5). The multi-way valve (8) is provided with at least an upper screen plate exhaust valve (a), a piston assembly internal exhaust valve (c), and a flow phase valve (b); the flow phase inlet / outlet pipe (b1) connected to the flow phase valve passes through the center of the piston assembly (1) and extends beyond the upper screen plate (11) at the bottom; the exhaust channel (a1) connected to the upper screen plate exhaust valve has its bottom flush with the center of the upper screen plate (11) and surrounds the flow phase inlet / outlet pipe; the exhaust channel (c1) connected to the piston assembly internal exhaust valve is located at the upper edge inside the piston assembly (1).

2. The chromatography column packing system according to claim 1, characterized in that, The lower distribution plate (4) has a central hole in the center. The lower distribution plate (4) faces downward. The lower distribution plate (4) has several long strip-shaped downward protruding distribution teeth (12) that radiate from the center hole toward the edge of the lower distribution plate (4). Between adjacent distribution teeth, there are half-distribution teeth (13) that radiate from the lower distribution plate (4) to the angle between adjacent distribution teeth. The lower distribution plate (4) has several through holes between adjacent distribution teeth and between distribution teeth and their adjacent half-distribution teeth.

3. The chromatography column packing system according to claim 1, characterized in that, The upper sieve plate (11) has a multi-layer mesh structure.

4. A method for packing a chromatography column based on the chromatography column packing system according to any one of claims 1-3, characterized in that, Inside the column, the packing material is dispersed, allowed to settle, vented, and compressed to complete the column loading process. The specific process is as follows: Step 1: Packing material dispersion: The packing material is homogenized in a homogenizing tank to obtain uniformly dispersed packing material. Step 2: Column packing: Pack the packing material that was broken up in Step 1 into the chromatography column. If the packing material settles naturally and forms a uniform column, the column packing is complete and a qualified column bed is formed. If there are air bubbles in the packing material or the column shape is not qualified, proceed to Step 3. Step 3: Inject buffer solution: Raise the piston assembly and inject buffer solution into the column at the same time during the raising process. After the column is filled with buffer solution, use compressed air to blow the column bed and the upper sieve plate to completely break up the column bed. The surface of the upper sieve plate needs to be kept dry. Step 4: Column bed sedimentation: After the packing material is evenly dispersed, it settles naturally, or the buffer solution is pumped into the upper end of the column and the buffer solution is pumped out from the lower end of the column to compact the column bed. Step 5: Column venting: First, close the gas / liquid mixing valve of the end cap assembly. The inside of the column is connected to the outside through the multi-port valve of the piston assembly. Press down the piston assembly, and the buffer solution will be discharged from the multi-port valve, bringing out the air at the same time. Then close the multi-port valve of the piston assembly and open the gas / liquid mixing valve of the end cap assembly. At this time, the inside of the column can only be connected to the outside through the gas / liquid mixing valve of the end cap assembly. Continue to press down the piston assembly, and the buffer solution will be discharged from the gas / liquid mixing valve of the end cap assembly, squeezing out the air bubbles inside the gas / liquid mixing valve of the end cap assembly. Step 6: Compress the packing: The lowering piston assembly compresses the packing, stopping when it reaches the target position according to the compressibility factor, thus completing the packing.

5. The column packing method for chromatography according to claim 4, characterized in that, In step 2, the packing is pumped into the column through the multi-port valve of the piston assembly, or drawn into the column through the gas / liquid mixing valve of the end cap assembly.

6. The column packing method for chromatography according to claim 4, characterized in that, In step 3, the upper screen plate is kept dry before venting. The method to keep the surface of the upper screen plate dry is to remove the upper screen plate from the liquid surface of the column before venting, and then blow the upper screen plate dry with compressed gas.

7. The column packing method for chromatography according to claim 4, characterized in that, In step 3, compressed air is blown into the packing column from the gas / liquid mixing valve of the end cap assembly and blown until the packing is uniform, and the pressure of the compressed air is not less than 1.5 Bar.

8. The column packing method for chromatography according to claim 4, characterized in that, In step 5, the air bubbles on the surface of the sieve plate are vented by pressing down the piston assembly, or by pumping buffer solution into the column while the piston assembly is stationary.

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