Method for removing bacteriophage endotoxin

Abstract

The invention relates to the technical field of biology, in particular to a method for removing bacteriophage endotoxin, which comprises the following steps: step 1, selecting ion exchange resin as an adsorption material; step 2, removal pretreatment: putting ion exchange resin into a chromatographic column, and washing and soaking the ion exchange resin in the chromatographic column with a 0.5-1M sodium hydroxide standard solution; and step three, removing endotoxin from the bacteriophage: adjusting the pH value of the ion exchange resin to 7.2-7.4, adding the bacteriophage of which the endotoxin is to be removed into the chromatographic column, and standing for 10-15 minutes. According to the method, the ion exchange resin is used as an adsorption material and is loaded into the chromatographic column to remove the phage endotoxin, other equipment is not needed, and the operation is simple; the ion exchange resin is high in durability and stability and can be repeatedly used and stably stored, so that the cost is reduced; meanwhile, the removal process is efficient and rapid.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, in particular to a method for removing toxins from bacteriophages. Background Art

[0002] Endotoxins are lipopolysaccharides (LPS) found in the cell walls of Gram-negative bacteria. They are released into the environment when the bacteria die and lyse. Bacteriophages, as viruses, do not inherently contain endotoxins. However, endotoxin contamination often occurs during the phage preparation process due to the lysis of the host bacteria. Currently, methods for removing phage endotoxins primarily include chemical, biological, and physical methods.

[0003] Existing methods for removing toxins from bacteriophages, such as ultrafiltration and gel chromatography, are costly and require high professional skills from operators. At the same time, existing low-cost methods include dialysis, but this method has a cumbersome operating process and requires more than 48 hours of removal time.

[0004] In summary, how to solve the problems of high cost, complicated operation and long time consumption has become a difficult problem that needs to be solved urgently in this field. Therefore, it is necessary to propose a low-cost, simple to operate and rapid method for removing toxins from bacteriophages. Summary of the Invention

[0005] To address the above-mentioned problems, the present invention provides a method for removing toxins from bacteriophages. Ion exchange resin is used as an adsorption material and loaded into a chromatography column to remove toxins from bacteriophages. This method does not require the use of other equipment and is simple to operate. The ion exchange resin is durable and stable, can be reused, and can be stably stored, reducing costs. Furthermore, the removal process is efficient and rapid, taking less than 2 hours.

[0006] In order to achieve the above object, the technical solution of the present invention is as follows: A method for removing toxins from bacteriophages, comprising the following steps:

[0007] Step 1: Select ion exchange resin as the adsorption material.

[0008] Step 2: Removal of pretreatment: Place the ion exchange resin into a chromatography column, and wash and soak the ion exchange resin in the chromatography column with a 0.5M-1M sodium hydroxide standard solution.

[0009] Step 3: Removal of endotoxins from phages: Adjust the pH of the ion exchange resin to 7.2-7.4, add the phages to be endotoxin-free to the chromatography column, and let it stand for 10-15 minutes.

[0010] Furthermore, in step 2, the preparation method of the 0.5M-1M sodium hydroxide standard solution is as follows: weighing 20-40g of sodium hydroxide solid into a beaker, adding 100-200mL of distilled water to the beaker, using a shaking component to dissolve the sodium hydroxide solid to obtain a sodium hydroxide solution and cooling it. After the sodium hydroxide solution is cooled, it is drained into a 1000mL volumetric flask with a glass rod.

[0011] Wash the beaker and glass rod 2-3 times with distilled water and transfer them to a volumetric flask. Continue adding distilled water to the volumetric flask. When the concave liquid surface of the sodium hydroxide solution in the volumetric flask is aligned with the scale line of the volumetric flask, stop adding distilled water to obtain a 0.5M-1M sodium hydroxide standard solution.

[0012] Furthermore, in step 2, the washing times are 2-3 times, and after washing, the mixture is soaked in a 0.5M-1M sodium hydroxide standard solution for more than 12 hours.

[0013] Furthermore, in step 3, the pH is adjusted by washing the ion exchange resin 3-4 times with 10x phosphate buffered saline at a flow rate of 0.5 mL / min to obtain an ion exchange resin with a pH of 7.2-7.4.

[0014] Furthermore, a 10x phosphate buffered saline solution is prepared by weighing 40-80 g of sodium chloride solid, 1-2 g of potassium chloride solid, 16.15 g-32.3 g of disodium hydrogen phosphate dodecahydrate solid, and 1.2 g-2.4 g of potassium dihydrogen phosphate solid; placing the weighed solids into a beaker in sequence, adding 300-800 mL of distilled water to the beaker, and using a shaking assembly to dissolve the solids in the beaker to obtain a phosphate buffered saline solution, and adjusting the pH of the phosphate buffered saline solution to 7.2-7.4 using hydrochloric acid.

[0015] Drain the pH-adjusted phosphate buffered saline solution into a 500-1000 mL volumetric flask with a glass rod. Wash the beaker and glass rod 2-3 times with distilled water and transfer them to the volumetric flask. Continue adding distilled water to the volumetric flask. Stop adding distilled water after the concave liquid surface of the phosphate buffered saline solution in the volumetric flask is aligned with the scale line of the volumetric flask to obtain a 10x phosphate buffered saline solution.

[0016] Furthermore, in step 3, when the phage is taken out after standing for 10-15 minutes, 2-3 ml of the front phage is discarded and the remaining phage is collected using a sterile, enzyme-free collection tube. The collection tube contains the phage with endotoxin removed.

[0017] Furthermore, the titer and activity of the phage in the collection tube were tested; both the titer and activity of the phage were tested using the double-plate method.

[0018] Furthermore, the chromatography column from which the phage endotoxins had been removed was washed 2-3 times with a 0.5M-1M sodium hydroxide standard solution, and the pH of the ion exchange resin in the chromatography column was adjusted to 7.2-7.4 with a 10x phosphate buffered saline solution. Two parts of anhydrous ethanol and eight parts of 0.1M sodium chloride solution were added and soaked until the next use.

[0019] Furthermore, the oscillation component includes a connecting shaft, and a plurality of sliding shafts are fixedly connected to the side wall of the connecting shaft in a circumferential direction. The sliding shafts are all slidably fitted with sliding plates, and the tops of the sliding plates are all fixedly connected to fixed plates.

[0020] A connecting port is provided at the top of the connecting shaft, and a connecting column is rotatably fitted in the connecting port. One end of the connecting column located at the upper part of the connecting shaft is fixedly connected to the base plate, and one end of the connecting column located at the lower part of the connecting shaft is fixedly connected to a driving member for driving the connecting column to rotate; fixed columns are fixedly connected to the fixed plates, and arc rods are hinged on the outer side walls of the fixed columns, and the ends of the arc rods away from the fixed columns are rotatably fitted with rotating shafts, and several rotating shafts are respectively fixedly connected to the top and bottom of the base plate.

[0021] The top of the base plate is fixedly connected with a fixed shaft, and a fixed cup is ball-hinged on the fixed shaft; the bottom of the connecting shaft is fixedly connected with a support box for protecting the driving component.

[0022] A protective component for limiting the tilt angle of the fixed cup is also provided on the top of the fixed plate.

[0023] Furthermore, the protection component includes a support column, which is fixedly connected to the top of the fixed plate, and a protection rod is fixedly connected to the side wall of the support column close to the connecting shaft.

[0024] The above scheme has the following beneficial effects:

[0025] 1. In the present invention, the ion exchange resin is used as an adsorption material for removing toxins from bacteriophages based on ion exchange. Bacteriophage endotoxins usually carry a negative charge, and the ion exchange resin can interact with the negative charge of the endotoxin to adsorb the endotoxin, thereby achieving the effect of removing the toxins from the bacteriophages. At the same time, the ion exchange resin has low adsorption to the bacteriophages themselves, which can ensure the number and activity of the bacteriophages.

[0026] Using a standard sodium hydroxide solution to wash and soak the ion exchange resin can remove impurities and pollutants on the surface of the ion exchange resin and ensure the stability of its adsorption performance. During the soaking process, sodium hydroxide can adjust the surface properties of the ion exchange resin, enhance its adsorption of endotoxins, and facilitate the removal of endotoxins.

[0027] The pH of the ion exchange resin was controlled at 7.2-7.4 using 10x phosphate buffered saline solution, which maintained its pH stability, avoided interference with the phage endotoxin adsorption process due to pH changes, and improved the endotoxin removal effect.

[0028] The titer and activity of phages are both detected using the double-plate method; this method is simple and highly accurate, and can accurately detect the quality of phages after the removal of phage toxins, thereby promptly discovering problems that occur during the phage toxin removal process.

[0029] 2. The present invention adds bacteriophages to a chromatography column with a pH controlled at 7.2-7.4 and allows the column to stand for 10-15 minutes to remove toxins from the bacteriophages. This method does not require any other equipment and is simple to operate. During the standing process, the endotoxins are in full contact with the ion exchange resin, thereby reducing the removal time and making the removal process more efficient and rapid.

[0030] When collecting phage after endotoxin removal, first discard 2-3 ml of front-end phage to prevent front-end phage that may be contaminated or has not been completely endotoxin-free from entering the collection tube, thereby ensuring the quality of the collected phage.

[0031] Ion exchange resins are durable and stable, allowing for repeated use and stable storage, reducing costs. Cleaning and soaking the ion exchange resin in a standard sodium hydroxide solution ensures stable storage for over three months and allows for over 50 cycles without compromising endotoxin removal, thus reducing costs. When not in use for extended periods, adjusting the pH and then soaking in a preservative solution can extend the lifespan of the chromatography column and ion exchange resin, ensuring stable and reliable performance.

[0032] 3. Before the present invention shakes the beaker of solution containing solid undissolved matter, the beaker is first placed in the fixed cup in the shaking assembly. The driving part in the shaking assembly drives the connecting shaft to rotate, the bottom plate to rotate, and the fixed shaft to rotate at the same time, so that the fixed cup rotates. The rotation of the beaker drives a vortex to be generated in the middle of the solution for stirring; at the same time, because the fixed cup and the fixed shaft are spherically hinged, the fixed cup will also tilt during the rotation of the fixed cup, driving the beaker to tilt and causing the liquid to continuously flow to one side, so that the solutions can more fully exchange positions with each other, accelerate the mixing of the solid undissolved matter and the solution, improve the uniformity of dissolution, ensure that the concentration of the configured solution is closer to the target concentration, and be more conducive to washing and soaking the ion exchange resin and adjusting the pH, thereby improving the removal effect of toxins in the bacteriophage.

[0033] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 The flowchart of the method for removing toxins from bacteriophages according to the present invention is shown.

[0035] Figure 2An axonometric view of an oscillating assembly of the method for removing toxins from bacteriophages according to the present invention.

[0036] Figure 3 An axonometric view of an oscillating assembly of the method for removing toxins from bacteriophages according to the present invention.

[0037] Figure 4 This is a cross-sectional view of a driving member of an oscillating assembly in the method for removing toxins from bacteriophages according to the present invention.

[0038] The figure marks in the drawings of the specification include: 1. support column; 2. protection rod; 3. sliding shaft; 4. sliding plate; 5. fixed plate; 6. fixed column; 7. arc rod; 8. connecting shaft; 9. connecting column; 10. rotating shaft; 11. fixed shaft; 12. bottom plate; 13. fixed cup; 14. motor; 15. support box. DETAILED DESCRIPTION

[0039] The following is further described in detail through specific implementation methods:

[0040] Example 1:

[0041] As attached Figure 1 A method for removing toxins from bacteriophages, comprising the following steps:

[0042] Step 1: Select ion exchange resin as the adsorption material.

[0043] Step 2: Removal of pretreatment: Place the ion exchange resin into a chromatography column, and wash and soak the ion exchange resin in the chromatography column with a 0.5 M sodium hydroxide standard solution.

[0044] The preparation method for a 0.5M sodium hydroxide standard solution is as follows: weigh 20g of solid sodium hydroxide into a beaker, add 100mL of distilled water to the beaker, and dissolve the solid sodium hydroxide using a shaking assembly to obtain a sodium hydroxide solution, which is then cooled. After the sodium hydroxide solution has cooled, drain it into a 1000mL volumetric flask using a glass rod. Rinse the beaker and glass rod three times with distilled water and transfer the solution to the volumetric flask. Continue adding distilled water to the volumetric flask until the concave meniscus of the sodium hydroxide solution in the volumetric flask aligns with the scale line on the volumetric flask, and stop adding distilled water to obtain a 0.5M sodium hydroxide standard solution. Wash three times, and then soak in a 0.5M sodium hydroxide standard solution for at least 12 hours.

[0045] Step 3: Phage Endotoxin Removal: Adjust the pH of the ion exchange resin to 7.2. Add the phage to be endotoxin-free to the chromatography column and let it stand for 15 minutes. Adjust the pH by washing the ion exchange resin four times with 10x phosphate buffered saline at a flow rate of 0.5 mL / min to obtain an ion exchange resin with a pH of 7.2.

[0046] A 10x phosphate buffered saline solution was prepared by weighing 40 g of solid sodium chloride, 1 g of solid potassium chloride, 16.15 g of solid disodium hydrogen phosphate dodecahydrate, and 1.2 g of solid potassium dihydrogen phosphate; placing the weighed solids into a beaker, adding 300 mL of distilled water to the beaker, and dissolving the solids in the beaker using a shaking assembly to obtain a phosphate buffered saline solution. The pH of the phosphate buffered saline solution was adjusted to 7.2 using hydrochloric acid.

[0047] The pH-adjusted phosphate buffered saline solution was drained into a 500 mL volumetric flask using a glass rod. The beaker and glass rod were washed three times with distilled water and transferred to the volumetric flask. Distilled water was then added to the volumetric flask. When the concave liquid surface of the phosphate buffered saline solution in the volumetric flask was aligned with the scale line of the volumetric flask, the addition of distilled water was stopped to obtain a 10x phosphate buffered saline solution.

[0048] When the phage is taken out after standing for 15 minutes, discard the front 3 ml of phage and use a sterile enzyme-free collection tube to collect the remaining phage. The collection tube contains the phage with endotoxin removed.

[0049] The titer and activity of the phage in the collection tube were tested; the titer and activity of the phage were both tested using the double-plate method.

[0050] After washing the chromatographic column from which phage toxins had been removed three times with a 0.5 M sodium hydroxide standard solution, the pH of the ion exchange resin in the chromatographic column was adjusted to 7.2 with a 10x phosphate buffered saline solution, and the column was soaked in 20% anhydrous ethanol and 80% 0.1 M sodium chloride solution until the next use.

[0051] The above method uses ion exchange resin as an adsorption material, which is loaded into a chromatography column to remove toxins from bacteriophages. It does not require the use of other equipment and can be completed in only two steps, making it easy to operate. The ion exchange resin is durable and stable, can be reused at least 50 times, and the single cost is less than 40 yuan. At the same time, it can be stably stored for at least 3 months, avoiding frequent replacement, reducing operating steps, and lowering costs. The removal process is efficient and fast, taking less than 2 hours.

[0052] This method greatly improves operational efficiency, provides a practical and efficient solution for quickly obtaining highly active, low-endotoxin-contaminated phages, and has effectively promoted the development of phage-related research and clinical applications.

[0053] Example 2:

[0054] The difference from the above embodiment is that, as shown in the attached Figure 2 -Attached Figure 4 As shown:

[0055] The oscillation assembly includes a connecting shaft 8 , on the side wall of which a plurality of sliding shafts 3 are circumferentially welded, each of which is slidably fitted with a sliding plate 4 , and a fixed plate 5 is welded on the top of each of which.

[0056] A connecting port is provided at the top of the connecting shaft 8, and a connecting column 9 is rotatably engaged in the connecting port. A base plate 12 is welded to one end of the connecting column 9 located at the upper part of the connecting shaft 8, and a driving member for driving the connecting column 9 to rotate is provided at one end of the connecting column 9 located at the lower part of the connecting shaft 8. In this embodiment, the driving member is a motor 14, and the output shaft of the motor 14 is fixedly connected to the lower end of the connecting column 9 with a coaxial bolt; fixed columns 6 are welded on the fixing plates 5, and arc rods 7 are hinged on the outer side walls of the fixing columns 6. The ends of the arc rods 7 away from the fixed columns 6 are rotatably engaged with rotating shafts 10, and several rotating shafts 10 are respectively welded to the top and bottom of the base plate 12.

[0057] A fixed shaft 11 is welded to the top of the bottom plate 12 , and a fixed cup 13 is spherically hinged on the fixed shaft 11 ; a support box 15 for protecting the motor 14 is fixedly connected to the bottom of the connecting shaft 8 by bolts.

[0058] A protective assembly is also provided on the top of the fixing plate 5 for limiting the tilt angle of the fixing cup 13. The protective assembly includes a support column 1, which is welded to the top of the fixing plate 5, and a protective rod 2 is welded on the side wall of the support column 1 near the connecting shaft 8.

[0059] The specific implementation process is as follows:

[0060] The experimenter places a beaker of solution containing undissolved solid matter into the fixed cup 13, turns on the motor 14 to rotate the connecting column 9 counterclockwise, the bottom plate 12 rotates, and the fixed shaft 11 rotates synchronously, so that the rotation of the fixed cup 13 drives the beaker to rotate counterclockwise; after rotating 1 / 4 of a circle, the motor 14 starts to rotate clockwise, causing the beaker to rotate clockwise as well. During the reciprocating rotation of the beaker, a rotating vortex is formed in the middle of the solution in the beaker. The vortex causes convection motion between the solutions, and the solution around the undissolved solid matter is continuously updated, thereby evenly dispersing the undissolved solid matter in the solution.

[0061] The fixed shaft 11 and the fixed cup 13 are connected by a ball hinge, so the fixed cup 13 will tilt during the rotation, and the beaker will also tilt accordingly. The tilt of the beaker causes the liquid to continuously flow to one side, so that the solutions exchange positions with each other, accelerating the mixing of solid undissolved matter and the solution; the rotation and tilting of the beaker cooperate with each other to make the position exchange of the liquid in the beaker more sufficient, improve the uniformity of dissolution, ensure that the concentration of the configured solution is closer to the target concentration, and be more conducive to washing and soaking the ion exchange resin and adjusting the pH, thereby improving the removal effect of toxins in the bacteriophage.

[0062] Tilting the beaker can also bring the residual solid undissolved matter on the wall of the beaker back into the solution, reducing the residue. At the same time, it prevents the solid undissolved matter from gathering on the wall of the beaker, making the concentration of the solution more uniform.

[0063] When preparing sodium hydroxide solution, heat is released during the dissolution of sodium hydroxide solid. The tilted beaker increases the contact area between the solution and the air, which can dissipate the generated heat in time, prevent the solution temperature from being too high, and ensure the safety of the experiment.

[0064] During the counterclockwise rotation of the base plate 12, the arc-shaped rods 7 rotate accordingly on the rotating shaft 10, exerting a pulling force on the fixed column 6 hinged at the end away from the rotating shaft 10, and the fixed plates 5 welded to the fixed columns 6 are also subjected to the same-direction pulling force, so that the sliding plate 4 slides toward the middle on the sliding shaft 3, and the support column 1 and the protective rod 2 move toward the middle accordingly; the protective rod 2 can limit the inclined fixed cup 13 to prevent the fixed cup 13 from tilting too much and causing the liquid in the beaker to flow out; at the same time, during the clockwise rotation, the support column 1 and the protective rod 2 move outward accordingly, which can continuously increase the tilt angle of the beaker, so that the solution is more dissolved and more uniform.

[0065] When the solid undissolved matter is dissolved, the motor 14 is turned off and the beaker is removed. The support box 15 can protect the motor 14 and play a role in supporting the oscillation component.

[0066] Obviously, the above embodiments are merely examples for clarity of explanation and are not intended to limit the implementation methods. Those skilled in the art will readily appreciate that other variations or modifications based on the above descriptions are possible. It is not necessary and impossible to enumerate all implementation methods here. Obvious variations or modifications arising therefrom remain within the scope of protection of the present invention.

Claims

1. A method for removing toxins from bacteriophages, characterized in that: The following steps are involved: Step 1, selecting ion exchange resin as adsorption material; Step 2: Removal of pretreatment: Place the ion exchange resin into a chromatography column and wash and soak the ion exchange resin in the chromatography column with a 0.5M-1M sodium hydroxide standard solution; Step 3: Removal of endotoxins from phages: Adjust the pH of the ion exchange resin to 7.2-7.4, add the phages to be endotoxin-free to the chromatography column, and let it stand for 10-15 minutes.

2. The method for removing bacteriophage endotoxins according to claim 1, characterized in that: In step 2, the preparation method of 0.5M-1M sodium hydroxide standard solution is: Weigh 20-40 g of solid sodium hydroxide into a beaker, add 100-200 ml of distilled water to the beaker, and use a shaking assembly to dissolve the solid sodium hydroxide to obtain a sodium hydroxide solution, which is then cooled. After the sodium hydroxide solution has cooled, drain it into a 1000 mL volumetric flask using a glass rod. Wash the beaker and glass rod 2-3 times with distilled water and transfer them to a volumetric flask. Continue adding distilled water to the volumetric flask. When the concave liquid surface of the sodium hydroxide solution in the volumetric flask is aligned with the scale line of the volumetric flask, stop adding distilled water to obtain a 0.5M-1M sodium hydroxide standard solution.

3. The method for removing phage endotoxins according to claim 2, characterized in that: In step 2, the washing times are 2-3 times, and after washing, the sample is soaked in a 0.5M-1M sodium hydroxide standard solution for more than 12 hours.

4. The method for removing phage endotoxins according to claim 3, characterized in that: In step 3, the pH is adjusted by washing the ion exchange resin 3-4 times with 10x phosphate buffered saline at a flow rate of 0.5 mL / min to obtain an ion exchange resin with a pH of 7.2-7.

4.

5. The method for removing phage endotoxins according to claim 4, characterized in that: The preparation method of 10x phosphate buffered saline is as follows: Weigh 40-80 g of sodium chloride solid, 1-2 g of potassium chloride solid, 16.15 g-32.3 g of disodium hydrogen phosphate dodecahydrate solid, and 1.2 g-2.4 g of potassium dihydrogen phosphate solid respectively; place the weighed solids into a beaker in sequence, add 300-800 mL of distilled water to the beaker, and use a shaking assembly to dissolve the solids in the beaker to obtain a phosphate buffered saline solution, and adjust the pH of the phosphate buffered saline solution to 7.2-7.4 with hydrochloric acid; Drain the pH-adjusted phosphate buffered saline solution into a 500-1000 mL volumetric flask with a glass rod. Wash the beaker and glass rod 2-3 times with distilled water and transfer them to the volumetric flask. Continue adding distilled water to the volumetric flask. Stop adding distilled water when the concave liquid surface of the phosphate buffered saline solution in the volumetric flask is aligned with the scale line of the volumetric flask to obtain a 10x phosphate buffered saline solution.

6. The method for removing phage endotoxins according to claim 5, characterized in that: In step 3, when the phage is taken out after standing for 10-15 minutes, 2-3 ml of the front phage is discarded and the remaining phage is collected using a sterile, enzyme-free collection tube. The collection tube contains the phage with endotoxin removed.

7. The method for removing phage endotoxins according to claim 6, characterized in that: The titer and activity of the phage in the collection tube were tested; both the titer and activity of the phage were tested using the double-plate method.

8. The method for removing phage endotoxins according to claim 7, characterized in that: After washing the chromatographic column from which phage toxins have been removed 2-3 times with a 0.5M-1M sodium hydroxide standard solution, adjust the pH of the ion exchange resin in the chromatographic column to 7.2-7.4 with a 10x phosphate buffered saline solution, and soak it in two parts of anhydrous ethanol and eight parts of 0.1M sodium chloride solution until the next use.

9. The method for removing phage endotoxins according to claim 8, characterized in that: The oscillation component comprises a connecting shaft (8), a plurality of sliding shafts (3) are fixedly connected to the side wall of the connecting shaft (8) in a circumferential direction, a sliding plate (4) is slidably fitted on each of the sliding shafts (3), and a fixed plate (5) is fixedly connected to the top of each of the sliding plates (4); A connecting hole is provided at the top of the connecting shaft (8), and a connecting column (9) is rotatably fitted in the connecting hole. One end of the connecting column (9) located at the upper part of the connecting shaft (8) is fixedly connected to the bottom plate (12), and one end of the connecting column (9) located at the lower part of the connecting shaft (8) is fixedly connected to a driving member for driving the connecting column (9) to rotate. The fixing plates (5) are all fixedly connected to fixing columns (6), and the outer side walls of the fixing columns (6) are all hinged with arc-shaped rods (7). The ends of the arc-shaped rods (7) away from the fixing columns (6) are all rotatably fitted with rotating shafts (10), and a plurality of rotating shafts (10) are respectively fixedly connected to the top and bottom of the bottom plate (12). The top of the bottom plate (12) is fixedly connected to a fixed shaft (11), and a fixed cup (13) is spherically hinged on the fixed shaft (11); the bottom of the connecting shaft (8) is fixedly connected to a support box (15) for protecting the driving member; A protective component for limiting the tilt angle of the fixing cup (13) is also provided on the top of the fixing plate (5).

10. The method for removing phage endotoxins according to claim 9, characterized in that: The protection assembly comprises a support column (1), the support column (1) is fixedly connected to the top of the fixed plate (5), and a protection rod (2) is fixedly connected to the side wall of the support column (1) close to the connecting shaft (8).

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