A solid-liquid separation method for 23-valent pneumococcal fermentation lysate

Abstract

The present invention provides a novel solid-liquid separation method for 23-valent pneumococcal fermentation lysate, using a cake layer candle solid-liquid separation system, in conjunction with a filter aid formulation that meets pharmaceutical grade standards, to clarify and filter the raw material liquid, replacing the existing two-step process: a process combination of a continuous flow disc separator + a deep filter, under the premise of the same quality index requirements for the clarity of the clarified liquid, shortening the process production process, simplifying the operating process and production system, improving the product yield, and achieving energy saving and consumption reduction, reducing production costs, and being environmentally friendly. In order to meet the high sanitation requirements of biopharmaceutical production equipment and the requirements of easy cleaning without dead corners, the present invention optimizes the equipment structure of the cake layer candle solid-liquid separation system, and makes corresponding designs for the operating procedures, so that the entire system meets the requirements of cleaning validation.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology and relates to a solid-liquid separation method of 23-valent pneumococcal fermentation lysate. Background Art

[0002] Solid-liquid separation is an essential production technology for the production of biological products. Sedimentation technology or filtration separation technology is used to effectively separate large particles in the fermentation lysate from the solution containing valuable components. The solid-liquid separation process is a necessary pre-treatment process for removing granular invalid substances for the subsequent further purification of valuable components.

[0003] At present, the solid-liquid separation of 23-valent pneumococcal fermentation lysate is mostly carried out by combining a continuous flow disc centrifuge with a deep filter. There are some problems in actual use:

[0004] A. Only using continuous flow disc centrifuge to process fermentation lysate will result in low clarity of clarified liquid and high proportion of particulate matter, which cannot meet the feed quality requirements of subsequent ultrafiltration processing. In order to meet the requirements of subsequent processes, it is necessary to use a deep filter for further processing to improve the clarity of the material to meet the feed requirements of ultrafiltration processing.

[0005] Ultrafiltration using a deep filter is a high-precision filtration process, and can only be used in the biopharmaceutical field in the form of expensive disposable single-use filters, which leads to a significant increase in production costs.

[0006] B. The combined solid-liquid separation method using a continuous flow disc centrifuge and a deep filter is used for solid-liquid separation of 23-valent pneumococcal fermentation lysate. It is a combination of two process method units, which lengthens the process production line, increases the production links, and makes the production more complex and difficult.

[0007] C. Due to the particularity of biopharmaceutical production, the cleaning-in-place (CIP) and sterilization-in-place (SIP) requirements of the production line must be verified by the reproducibility of the results. The longer production process increases the complexity of the system and the difficulty of implementation, which increases the potential production quality risks, increases the difficulty of controlling production control stability, and greatly increases the investment in production equipment.

[0008] D. Due to the limitation of the working principle of the continuous flow disc centrifuge, there is always a certain amount of material that cannot be processed at the end, and complete batch filtration cannot be achieved, resulting in a lower product yield. This is undoubtedly a very high price to pay for a high value-added product such as the 23-valent pneumococcal vaccine.

[0009] E. Due to the limitation of the working principle of the continuous flow disc centrifuge, the separated granular residue can only be discharged in the form of fluidization. The problem is that due to the biosafety requirements, this part of the material must be inactivated. The fluidized solid phase residue has a large volume, and the difficulty and cost of processing are very high. Summary of the invention

[0010] The object of the present invention is to provide a separation method for solid-liquid separation of pneumococcal fermentation lysate.

[0011] The present invention is a solid-liquid separation technology for the fermentation lysate of 23-valent pneumococcal polysaccharide vaccine, which refers to the material after fermentation, culture, inactivation and lysis of pneumococci of various valent types. By using the method of the present invention, the process production process can be shortened under high-sanitary conditions, and complete batch solid-liquid separation can be achieved.

[0012] The present invention mainly uses the solid-liquid separation of the inactivated pneumococcal fermentation broth, uses a filter aid that meets pharmaceutical grade standards and a filter cake layer formed by tissues such as cell fragments and cell walls in the fermentation broth, to clarify and filter the raw liquid, so as to achieve the purpose of meeting the clarification index (turbidity ≤ 10NTU) that can be normally produced by the subsequent ultrafiltration process.

[0013] In order to achieve the above object, the present invention provides a solid-liquid separation method of 23-valent pneumococcal fermentation lysate, comprising the following steps:

[0014] Step 1: Select the filter aid formula: select the corresponding formula according to the valence type of the pneumonia vaccine fermentation lysate to be processed;

[0015] Step 2: Prepare a filter aid, and evenly mix the formulated filter aid with the fermentation lysate of the pneumonia vaccine to be processed in a feed tank; the formulated filter aid and the solids to be removed in the fermentation lysate of the pneumonia vaccine to be processed form large solid particles in a cross-linked state;

[0016] Step 3: Filter the mixed material in step 2 through a sanitary filter cake layer filter, which contains a filter cloth and a filter element; after filtration, the filtrate flows back to the feed tank, and the large solid particles in the cross-linked state contained in the mixed material in step 2 are intercepted by the filter cloth, forming a filter cake layer on the surface of the filter cloth; the filter cake layer constructs a flow channel due to the cross-linking bridging effect of the solid particles, allowing liquid and small solid particles to pass through; and as the filter cake layer thickens, the particle size of the solid particles allowed to pass through the filter cake layer flow channel becomes smaller, until the turbidity of the filtrate reaches the filtration target parameter;

[0017] Step 4: stop the filtrate from flowing back to the feed tank, transfer the filtrate to the filtrate tank, and obtain filtrate;

[0018] Step 5: Treatment of residual tailings: The remaining materials in the sanitary filter cake layer filter are controlled by air pressure to intercept the solids into the filter cake layer;

[0019] Step 6: washing the filter cake;

[0020] Step 7: Filter cake unloading: let the filter cake layer on the filter cloth fall off into the closed slag discharge mechanism;

[0021] Step 8: Transfer of filtrate: Transfer all filtrate to the ultrafiltration department.

[0022] Preferably, the filter aid formulation in step 1 comprises 1 # Diatomaceous earth, 2 # Fiber and 3 # Silicon dioxide powder.

[0023] Filter aids specifically refer to filter aids that meet the national requirements for pharmaceutical excipients, including but not limited to diatomaceous earth, cellulose, crystalline cellulose, silica particles, microsphere resin, etc.

[0024] Any one of the above is preferably that the 1 # The diatomaceous earth is composed of Celpure C1000 and Celpure C65 diatomaceous earth of Yiruishi, and the mass ratio is Celpure C1000: Celpure C65=8:2.

[0025] Any of the above is preferably that the 2 # The cellulose is at least one of the ARBOCEL series celluloses ARBOCEL15-18 from Ridenmeier.

[0026] Any one of the above is preferably that the 3 # The silicon dioxide powder is the silicon dioxide powder of model LH102 from Longhua Chengxin Powder Technology Co., Ltd.

[0027] Any of the above is preferably, in the valence types 1, 2, 3, 4, 5, 6A, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F, 33F of pneumococcus, the formula is: Formula 1: 1 # Diatomaceous earth: 2 # Fiber: 3 # The mass ratio of silicon dioxide powder is 6:2:1, which is suitable for solid-liquid separation of 1, 2, 6A, 7F, 9N, 9V, 10A, 11A, 12F, 17F, 19A, 19F, and 33F valence types of pneumococcus.

[0028] Any of the above is preferably, formula 2: 1 # Diatomaceous earth: 2 # Fiber: 3# The mass ratio of silicon dioxide powder is 5.5:2:1.5, which is suitable for solid-liquid separation of 4, 5, 22F and 23F valence types of pneumococcus.

[0029] Any of the above is preferably, formula three: 1 # Diatomaceous earth: 2 # Fiber: 3 # The mass ratio of silica powder is 6:3:1, which is suitable for solid-liquid separation of pneumococcal 14, 15B, 18C, and 20 valent forms.

[0030] Any of the above is preferably, formula 4: 1 # Diatomaceous earth: 2 # Fiber: 3 # The mass ratio of silicon dioxide powder is 7:2:1; it is suitable for solid-liquid separation of 3- and 8-valent forms of pneumococcus.

[0031] The present invention forms targeted and optimized filter aid ratios and use methods for pneumococcal fermentation lysates of different valences, respectively, to achieve high-efficiency, low-cost, and short-process line solid-liquid separation.

[0032] Any one of the above is preferably that the specific steps of step 2 are:

[0033] Step 2.1: inject the fermentation liquid to be treated into the feed tank and start stirring;

[0034] Step 2.2: Weigh and prepare the filter aid according to the formula, add the prepared filter aid into the feed tank under bacteria-controlled and closed conditions, and mix it evenly with the fermentation lysate of the pneumonia vaccine to be treated in the tank under stirring.

[0035] Any one of the above is preferably that the specific steps of step 3 are:

[0036] Step 3.1: Feeding and filling: conveying the mixed material in step 2 to a sanitary filter cake layer filter; the filter element is a sanitary perforated plum blossom filter element;

[0037] Step 3.2: Pre-coat filtration, filter cake layer is formed, and the filtrate reaches the process index: the material to be processed is filtered through the filter cloth and the filter element, so that the filtrate flows back to the feed tank; the large solid particles in the cross-linked state formed in the material to be processed are intercepted by the filter cloth, and a filter cake layer is formed on the surface of the filter cloth. The flow channel constructed by the cross-linking bridging effect of the solid particles in the filter cake layer allows liquid and some small solid particles to pass through; under the action of a pump pressure of 0.3 to 0.35MPa, the filter cloth is pressed tightly against the filter element, and the cross-section is plum blossom-shaped.

[0038] Any of the above is preferably that in step 4, when the filtrate standard meets the turbidity requirement of the filtration target, the filtrate is controlled by a valve to enter the filtrate tank to obtain a filtrate;

[0039] Any of the above items is preferably that step 5: treatment of residual tailings: the sanitary filter cake layer filter is provided with a distributor, the remaining material is sprayed out from the distributor, the air is compressed in the sanitary filter cake layer filter to form a gas-liquid-solid mixture, the pressure in the sanitary filter cake layer filter is controlled to be 0.3 to 0.35MPa, so that the gas-liquid-solid mixture passes through the filter cake layer and the filter cloth under the action of pressure, the solids are intercepted by the filter cake layer, the liquid enters the filter liquid tank, and the gas is discharged into the exhaust system through the filter liquid tank exhaust valve; this process continues until all materials in the sanitary filter cake layer filter are completely processed.

[0040] The present invention adopts the method of pressurizing the residual materials in the filter that cannot be filtered normally at the filtering end after being discharged, and re-enters the filter for filtering through a special feed material distribution device, thereby achieving complete processing of all materials and complete batch filtration, without any unusable or discarded raw materials, thereby improving product yield and reducing losses. Since the main filtering equipment is a static equipment, there is no power consumption and consumable parts and materials, thereby reducing power consumption as well as manpower and costs for maintenance.

[0041] Since the entire solid-liquid separation method production realizes the production operation under uninterrupted continuous control throughout the whole process, and realizes batch automated production under PLC (programmable logic controller) control, without human intervention, the traceability of actions and production data in each production link in the drug production process is reliably guaranteed to eliminate interference from human factors.

[0042] Programmable logic controller (PLC) is a digital computing and operating electronic system. The present invention adopts a conventional PLC in the prior art. The present invention preferably selects a corresponding formula function package on the PLC in step 1 according to the valence type of the fermentation lysate of the pneumonia vaccine to be processed.

[0043] Any one of the above is preferably that the specific steps of step 6 are:

[0044] Step 6.1: Fill the feed tank with water at a temperature of 20°C. Inject water to preliminarily wash the feed tank, and use the wash water to wash the filter cake;

[0045] Step 6.2: Washing filter cake: The washing water in the feed tank is transported to the sanitary filter cake layer filter to dilute the previous residual material in the filter. After the feed tank is emptied, repeat step 5 to treat the remaining tailings, and spray the filter cake with washing water until all the washing water in the sanitary filter cake layer filter enters the filtrate tank.

[0046] Any of the above items is preferably that in step 7, a closed slag discharge mechanism is connected below the sanitary filter cake layer filter, and by controlling the direction of the airflow in the sanitary filter cake layer filter, compressed air causes the air flow to act on the inside of the filter cloth and the filter element from downstream to upstream, and the filter cloth with a plum blossom-shaped cross-section is expanded into a circular shape under pressure, so that cracks appear in the plum blossom depression of the filter cloth, and the filter cake on the filter cloth falls off the filter cloth and falls to the closed slag discharge mechanism.

[0047] Any of the above items is preferably that in step 8, after the filter cake is washed in step 6.2, the filtrate is transferred to the ultrafiltration department.

[0048] Preferably, any of the above items further comprises a cleaning in place (CIP) step and a steam sterilization in place (SIP) step.

[0049] Any of the above is preferred, and the preferred filter cake layer candle type solid-liquid separation system of the present invention is provided. In the preferred embodiment 1 of the present invention, the preferred process of the present invention using the filter cake layer candle type solid-liquid separation system is provided.

[0050] The present invention adopts a special design to achieve the closed discharge of solid slag under the conditions of high sanitation level cleaning in place (CIP) and sterilization in place (SIP) technical requirements (the technology of Chinese utility model patent CN218188299U is creatively applied to the present invention, the application number of the patent is CN202222762731.2, application date: October 19, 2022; publication number: CN218188299U, publication date: January 3, 2023). Biological contamination is avoided, the amount of waste slag that needs to be biologically inactivated is greatly reduced, and the discharge amount is effectively reduced without polluting the clean operating environment.

[0051] In order to meet the high sanitation requirements of biopharmaceutical production equipment and the requirements of no dead corners and easy cleaning, the core equipment of the cake layer candle solid-liquid separation system, the cake layer candle filter, has made significant optimization and upgrade design:

[0052] A. The core optimization design content of the present invention is as follows: the present invention uses a new filter element as a special integrally formed filter element for biopharmaceutical production for the first time. The filter element is recorded in the Chinese utility model patent CN218901009U, the patent application number is CN202223038887.2, application date: November 11, 2022; publication number: CN218901009U, publication date: April 25, 2023. In the prior art, the process combination of "continuous flow disc separator + deep filter" is adopted, and the candle filter element is used, which requires two sets of equipment and two processes. In the preparation process, there are many cleaning dead corners, which cannot meet the technical defects of high-level sanitary biopharmaceutical equipment with no dead corners and easy cleaning.

[0053] The present invention is a one-step method with fewer steps and simplified equipment, so there are fewer cleaning dead corners. In addition, the filter element structure is easier to clean and is not prone to cleaning dead corners and residues. However, before the present invention, the filter element has not been applied to the high-sanitation biopharmaceutical equipment in the prior art. The present invention uses the cross-linking bridging effect of the solid particles formed by the filter aid and the fermentation lysate to make the filter element creatively applied to the high-sanitation biopharmaceutical equipment provided by the present invention. In the method of the present invention, the formula filter aid described in step 1 forms large solid particles in a cross-linked state with the solids to be removed in the fermentation lysate of the pneumonia vaccine to be treated, and the large solid particles in a cross-linked state contained in the mixed material in step 2 are intercepted by the filter cloth to form a filter cake layer on the surface of the filter cloth; the filter cake layer constructs a flow channel due to the cross-linking bridging effect of the solid particles, allowing liquid and small solid particles to pass through; and as the filter cake layer thickens, the particle size of the solid particles allowed to pass through the filter cake layer flow channel becomes smaller until the turbidity of the filtrate reaches the filtration target parameter. In the present invention, the gradually formed filter cake layer not only does not cause blockage of the filter element and reduce the performance of the filter element, but the filter cake layer and the filter element cooperate with each other to jointly achieve a filtering effect. The combination of the filter cake layer and the filter element improves the filter element to be more suitable for the requirements of high-sanitary biopharmaceutical equipment, so that the turbidity of the clear liquid after filtration can meet the requirements of high-sanitary biopharmaceuticals.

[0054] B. In order to apply the filter element to high-sanitary biopharmaceutical equipment, the entire production process method is adjusted. In a further improvement plan, the design of all pipelines in contact with materials, cleaning liquids and injection-grade water in the cake layer candle solid-liquid separation system of the present invention, the selection of materials and polishing grades are all optimized, and the dead zone of all pipelines shall not be greater than 2-3 times the inner diameter (2D, 3D principle), so that it fully meets the high-sanitary technical requirements of no dead angle and easy cleaning. For the parts that cannot meet the 2D and 3D principles due to hardware specification limitations, adjustments are made in the control program so that they can meet the cleaning requirements.

[0055] C. In a further improvement scheme, the formula of the filter aid is adjusted. The filter aid of the present invention is easier to form large solid particles in a cross-linked state with the solids to be removed in the fermentation lysate of the pneumonia vaccine to be treated, which is beneficial to the formation of a cross-linked bridging effect in the filter cake layer. The combined use of the further improved filter aid formula of the present invention and the filter element further improves the performance of the solid-liquid separation method provided by the present invention for clarifying and filtering the raw liquid, so that under the premise of the same quality index requirements of the clarity of the clarified liquid as the prior art, the method of the present invention replaces the two-step process in the prior art (continuous flow disc separator + deep filter process combination). At the same time, the filter aid formula of the present invention forms targeted optimized filter aid ratios and usage methods for pneumococcal fermentation lysates of different valences, respectively, to achieve high-efficiency, low-cost, short-process line solid-liquid separation.

[0056] D. In a further improvement scheme, through the method of step 5, the present invention adopts the method of pressurizing after exporting the residual materials in the filter that cannot be filtered normally at the end of filtration, and re-enters the filter for filtration through a special feed material distribution device, so that all materials can be completely processed and complete batch filtration can be achieved, without unusable and discarded raw materials, thereby improving product yield and reducing losses; because the main filtering equipment is a static equipment, there is no power consumption and loss-making parts and materials, which reduces power consumption and maintenance manpower and costs. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] Figure 1 This is a novel filter cake layer candle type solid-liquid separation system for biopharmaceutical production adopted in the preferred embodiment 1 of the present invention.

[0058] Figure 2 This is a structural diagram of the filter cake layer candle type solid-liquid separation system used in the preferred embodiment 1 of the present invention.

[0059] Figure 3 This is a schematic diagram of the working state of the filter element assembly in the cake layer candle type solid-liquid separation system adopted in the preferred embodiment 1 of the present invention.

[0060] Figure 4 Schematic diagram of the solid-liquid separation method of pneumococcal fermentation lysate in the prior art provided in the examples.

[0061] Figure 5 This is a schematic diagram of the method flow in preferred embodiment 1 of the present invention.

[0062] Figure 6 This is a diagram showing the effect of solid-liquid separation of the inactivated pneumococcal fermentation broth in the preferred embodiment 1 of the present invention.

[0063] Markings in the attached drawings:

[0064] Feed tank 001, filter aid 002, dosing port 003, material delivery pump 004, sanitary filter cake layer filter 005, feed tank respirator 006, filter cloth 008, filter element 009, filter element support orifice layer 091, filter element central tube wall 092, filter cake layer 010, turbidity meter 011, filter liquid tank 012, low liquid level switch 013, filter machine respirator 014, distributor 015, upstream pressure sensor 016, downstream pressure sensor 017, gas-liquid separator 018, filter liquid delivery pump 020, filter liquid tank level sensor 021, conductivity meter 022;

[0065] Material valve 101, material valve 2 102, tank bottom valve 103, material connecting valve 104, filter feed valve 105, filtrate outlet valve 106, filtrate valve 108, filtrate tank bottom valve 109, filtrate connecting valve 110, filtrate discharge valve 111, discharge valve 112, exhaust overflow valve 113, exhaust overflow valve 2 114, pre-coat reflux valve 115, return valve 116, tail valve 117, valve 118;

[0066] Cleaning main valve 201, injection water main valve 202, cleaning branch valve 203, cleaning branch valve 204, cleaning branch valve 205, upper cleaning ball valve 206, lower cleaning ball valve 207, cleaning branch valve 208, filtrate tank cleaning ball valve 209, valve 210, valve 211, cleaning branch valve 212, feed tank cleaning valve 213, cleaning branch valve 214, valve 215, cleaning branch valve 216, cleaning branch valve 217;

[0067] Steam main valve 301;

[0068] Feed tank air inlet valve 401, filter reverse air inlet valve 402, filter forward air inlet valve 403;

[0069] Feed tank exhaust valve 501, filtered liquid tank exhaust valve 502, filter exhaust valve 503;

[0070] Feed tank drain valve 701, steam trap 702, steam trap 703, filter drain valve 704, filtrate tank drain valve 705, steam trap 706, post-pump sewage valve 707, filtrate sewage main valve 708, steam trap 709, filtrate drain valve 710, wastewater main valve 712, waste alkali liquid main valve 713. DETAILED DESCRIPTION

[0071] The present invention is further described in detail below through the accompanying drawings and specific embodiments.

[0072] Example 1

[0073] In Example 1, a filter cake layer candle type solid-liquid separation system is used and the operation is performed according to the following method steps:

[0074] Step 1: Select a recipe.

[0075] According to the valence type of the pneumonia vaccine fermentation lysate to be processed, the corresponding formula function package is selected on the PLC.

[0076] Step 2: Preparation and addition of filter aid.

[0077] 2.1 About 600L of fermentation lysate to be treated is injected into the feed tank 001 through material valve 101 and material valve 2 102 with compressed air, and the agitator is turned on at the same time;

[0078] 2.2 Weigh and prepare the filter aid according to the formula. Add the prepared filter aid 002 into the feed tank 001 through the addition port 003 under bacteria control and closed conditions. Under the action of stirring, mix it evenly with the material to be treated in the tank. At this time, the formulated filter aid and the solid matter to be removed in the fermentation broth of this type (the main component is cell debris) form a certain cross-linking state.

[0079] Step 3: Pre-coat filtration.

[0080] 3.1 Feeding and filling: Open the material delivery pump 004, and deliver the mixed material to the sanitary cake filter 005 through the tank bottom valve 103, the material connecting valve 104, and the filter feed valve 105. At the same time, the gas in the sanitary cake filter 005 is discharged into the tail gas system through the exhaust overflow valve 113, the exhaust overflow valve 2 114, the material valve 2 102, the feed tank respirator 006, the feed tank intake valve 401 and the feed tank exhaust valve 501 to balance the pressure;

[0081] 3.2 Pre-coat filtration, filter cake layer is formed, and the filtrate reaches the process index: when the feed liquid is full, the high liquid level switch 007 gives a signal to control the closing of the exhaust overflow valve 113 and the exhaust overflow valve 2 114, and the filtrate outlet valve 106 and the pre-coat reflux valve 115 are opened, and the material to be processed passes through the filter cloth 008 and the sanitary perforated plum blossom filter element 009 (the structure of the sanitary perforated plum blossom filter element 009 is recorded in the Chinese utility model patent ZL202223038887.2, the patent application number: CN202223038887.2, application date: November 11, 2022; publication number: CN218901009U, publication date: April 25, 2023) is filtered, and the filtrate flows back to the feed tank 001, while the large solid particles (mixture of formula filter aid and cell fragments) in the material to be treated are intercepted by the filter cloth 008, and gradually form a filter cake layer 010 on the surface of the filter cloth. Due to the cross-linking and bridging effect of the solid particles, the filter cake layer constructs a relatively uniform flow channel, which allows liquid and some small solid particles to pass through. Under the action of 0.3--0.35MPa pump pressure, the filter cloth 008 is pressed tightly against the filter element 009, and the cross-section is plum blossom-shaped. The filter cake layer is relatively thin at the beginning of its formation, the flow channel is simple, and more solid particles can pass through. As the filtration process proceeds, the filter cake layer gradually thickens, the flow channel becomes more complex and dense, the filtration efficiency is improved, and the particle size of solids allowed to pass through becomes smaller and smaller, thereby achieving the filtration target parameter--turbidity 8--10NTU. At this time, the online turbidity meter 011 on the filtrate pipeline gives a signal to enter the next step.

[0082] Step 4: Collect the filtrate.

[0083] Open the filtered liquid valve 108, close the pre-coat reflux valve 115, and the filtered liquid enters the filtered liquid tank 012.

[0084] Step 5: Disposal of remaining tailings.

[0085] When the material in the feed tank 001 is emptied, the low liquid level switch 013 gives a signal, closing the material connecting valve 104 and the filter feed valve 105, and at the same time opening the return valve 116, the tail valve 117, the filter respirator 014, and the filter forward air inlet valve 403, so that the remaining material in the sanitary filter cake layer filter 005 is ejected from the distributor 015, and a gas-liquid-solid mixture is formed under the joint action of the compressed air in the filter. At this time, the pressure in the sanitary filter cake layer filter 005 is about 0.3-0.35MPa (upstream pressure sensor 016), that is, the pressure upstream of the filter cake layer 010, and the downstream of the filter cake layer 010 maintains a slight positive pressure (downstream pressure sensor 017), so the gas-liquid-solid mixture passes through the filter cake layer 010 and the filter cloth 008 under the action of pressure, and the solids therein are intercepted by the filter cake layer 010, and the gas-liquid mixture enters the filtrate pipe, and is further separated through the filter liquid valve 108 and the gas-liquid separator 018, and the liquid enters the filter liquid tank 012, and the gas is discharged into the exhaust system through the filter liquid tank exhaust valve 502. This process continues until all the materials in the sanitary filter cake layer filter 005 are basically processed. Since the feed pipeline after the filter feed valve 105 is connected to the bottom of the sanitary filter cake layer filter 005, the residual material volume does not exceed 2L, which is equivalent to 0.3% of the volume of the entire batch of feed liquid, which meets the requirements of the GMP2010 version for complete batch processing, and also ensures the product yield.

[0086] Step 6: Wash the filter cake.

[0087] 6.1 Filling the feed tank with water: Cool the injection water to 20°C, open the injection water main valve 202, the cleaning branch valve 212 and the feed tank cleaning valve 213 to inject a certain amount of injection water (about 100L), and use the washing ball to perform preliminary washing on the feed tank. At the same time, this washing water will be used as the source of washing the filter cake;

[0088] 6.2 Washing filter cake: Open the material delivery pump 004, and deliver the washing water in the feed tank to the sanitary filter cake layer filter 005 through the tank bottom valve 103, the material connecting valve 104, and the filter feed valve 105 to dilute the previous residual material in the filter (not exceeding 2L). After the feed tank 001 is emptied, repeat operation 5 to deal with the remaining tailings, and spray the filter cake with washing water through the distributor 015 until all the washing water in the filter is cleaned and enters the filtrate tank 012 to elute the effective ingredients of the product entrained in the filter cake, thereby further improving the yield.

[0089] Step 7: Filter cake unloading.

[0090] Connect the closed slag discharge mechanism 019 below the sanitary filter cake layer filter 005, release the pressure of the sanitary filter cake layer filter 005 to normal pressure through the filter respirator 014 and the filter exhaust valve 503, open the discharge valve 112 and the filtrate outlet valve 106, open the filter reverse air inlet valve 402 for a short time (3 seconds each time, 3 times in total) to allow the compressed air to act on the filter cloth 008 and the filter element 009 from the downstream to the upstream direction, and the filter cloth 008 with a plum blossom cross-section is expanded into a round shape under pressure, so that the filter cake on it has regular cracks in the plum blossom depression, and it smoothly detaches from the filter cloth and falls into the closed slag discharge mechanism. The reverse air is discharged to the exhaust system through the filter respirator 014 and the filter exhaust valve 503.

[0091] Step 8: Transfer the filtrate to another material.

[0092] Step 6.2 gives a signal when the filter cake washing is finished, opens the filter liquid tank bottom valve 109, the filter liquid connecting valve 110, and the filter liquid discharge valve 111, sends a communication signal to the downstream ultrafiltration department to request to receive the filter liquid, and after the ultrafiltration department is ready to receive the reply signal, starts the filter liquid delivery pump 020, transfers all the filter liquid to the ultrafiltration department, waits for the pump to be emptied, and the filter liquid tank level sensor 021 gives a signal, opens the filter machine reverse air inlet valve 402 and the filter liquid valve 108, and uses compressed air to clean the residual filter liquid in the pipeline.

[0093] Step 9: Cleaning in place (CIP).

[0094] 9.1 Wash with purified water.

[0095] 9.1.1 Confirmation of purified water: Send a communication signal to the CIP station to request cleaning water, open the main cleaning valve 201, and start the cleaning process when the pressure value meets the conditions;

[0096] 9.1.2 Cleaning the incoming material pipeline: open the material valve 101 and the cleaning branch valve 212, and send a cleaning signal for the incoming material pipeline to the fermentation department. After receiving the signal, the fermentation department opens the drainage pipeline and cleans the incoming material pipeline according to the preset time;

[0097] 9.1.3 Cleaning feed tank 001: Open the cleaning branch valve 212, the feed tank washing ball valve 213, the feed tank bottom valve 103, the material connecting valve 104, the feed tank drain valve 701, and the wastewater main valve 712 to clean the inside of the feed tank and the discharge pipeline. When the preset cleaning time is up, open the feed tank air inlet valve 401 to discharge the wastewater to the sewage station;

[0098] 9.1.4 Cleaning the sanitary cake layer filter 005: Open the cleaning branch valve 203, cleaning branch valve 204, cleaning branch valve 205 and the filter upper wash ball valve 206, lower wash ball valve 207, filter drain valve 704, and wastewater main valve 712 to wash the inside of the filter. Open the cleaning branch valve 214, cleaning branch valve 216, cleaning branch valve 217, filter feed valve 105, and open the material delivery pump 004 to wash the feed pipeline. Open the cleaning branch valve 212, pre-coating reflux valve 115, and filtrate outlet valve 106 to reversely clean the filter cloth 008 and filter element 009. Open the second material valve 102 to clean the pre-coating reflux pipeline. When the preset cleaning time is up, open the filter forward air inlet valve 403 to discharge the wastewater to the sewage station;

[0099] 9.1.5 Cleaning of the filter liquid tank 012: Open the cleaning branch valve 203, cleaning branch valve 204, filter liquid tank ball valve 209, filter liquid tank bottom valve 109, filter liquid tank drain valve 705 to clean the inside of the filter liquid tank and the discharge pipeline. When the preset cleaning time is up, close the filter liquid tank drain valve 705, open the filter liquid delivery pump 020, filter liquid connecting valve 110, filter liquid sewage main valve 708, filter liquid drainage valve 710, and wastewater main valve 712 to clean the filter liquid delivery pipeline, discharge the wastewater to the sewage station, empty the pump, and open the pump back sewage valve 707 to discharge the residual liquid.

[0100] The above steps are performed in sequence according to the cleaning time and step sequence settings.

[0101] 9.2 Clean with sodium hydroxide solution.

[0102] 9.2.1 Sodium hydroxide solution confirmation: Send a communication signal to the CIP station to request 0.5% sodium hydroxide solution (hereinafter referred to as "alkali solution"), open the cleaning main valve 201, and start the cleaning procedure when the pressure value meets the conditions;

[0103] 9.2.2 Cleaning the incoming material pipeline: Open the material valve 101 and the cleaning branch valve 212 to let in the alkali solution, and send a signal requesting cleaning of the incoming material pipeline to the fermentation department. After receiving the signal, the fermentation department opens the drainage pipeline and cleans the incoming material pipeline according to the preset time;

[0104] 9.2.3 Cleaning feed tank 001: Open the cleaning branch valve 212, the feed tank washing ball valve 213, the feed tank bottom valve 103, and the material connecting valve 104 to allow alkali solution to wash the inside of the feed tank. When the preset cleaning time is up, open the material delivery pump 004, open the filter feed valve 105 and the tail valve 117, and discharge the alkali solution to the sanitary filter cake layer filter 005;

[0105] 9.2.4 Cleaning of sanitary cake filter 005: Open the cleaning branch valve 212, pre-coating reflux valve 115, and filtrate outlet valve 106, and reversely clean the filter cloth 008 and filter element 009. Open the material valve 102 to clean the pre-coating reflux pipeline. When the preset cleaning time is up, open the filter forward air inlet valve 403, the filter drain valve 704, and the waste alkali liquid main valve 713 to press the waste alkali liquid to the waste alkali liquid recovery station;

[0106] 9.2.5 Cleaning the filter liquid tank 012: Open the cleaning branch valve 203, the cleaning branch valve 204, and the filter liquid tank washing ball valve 209 to pass the alkali solution to wash the filter liquid tank. When the preset cleaning time is up, open the filter liquid tank bottom valve 109, the filter liquid connecting valve 110, open the filter liquid delivery pump 020, the filter liquid sewage main valve 708, and the waste alkali liquid main valve 713 to discharge the waste alkali liquid to the waste alkali liquid recycling station, empty the pump, and open the pump back sewage valve 707 to discharge the residual liquid.

[0107] The above steps are performed in sequence according to the cleaning time and step sequence settings.

[0108] 9.3 Wash with water for injection.

[0109] 9.3.1 Confirmation of water for injection: Send a communication signal to the water for injection station to request water for injection, open the main valve 202 of water for injection, and start the cleaning procedure when the pressure value meets the conditions;

[0110] 9.3.2 Cleaning the incoming material pipeline: Open the material valve 101 and the cleaning branch valve 212, and send a signal requesting cleaning of the incoming material pipeline to the fermentation department. After receiving the signal, the fermentation department opens the drainage pipeline and cleans the incoming material pipeline according to the preset time;

[0111] 9.3.3 Cleaning feed tank 001: Open the cleaning branch valve 212, the feed tank washing ball valve 213, the feed tank bottom valve 103, the material connecting valve 104, the feed tank drain valve 701, and the wastewater main valve 712 to clean the inside of the feed tank and the discharge pipeline. When the preset cleaning time is up, open the feed tank air inlet valve 401 to discharge the wastewater to the sewage station;

[0112] 9.3.4 Cleaning the sanitary cake layer filter 005: Open the cleaning branch valve 203, cleaning branch valve 204, cleaning branch valve 205 and the filter upper wash ball valve 206, lower wash ball valve 207, filter drain valve 704, and wastewater main valve 712 to wash the inside of the filter. Open the cleaning branch valve 214, cleaning branch valve 216, cleaning branch valve 217, filter feed valve 105, and open the material delivery pump 004 to wash the feed pipeline. Open the cleaning branch valve 212, pre-coating reflux valve 115, and filtrate outlet valve 106 to reversely clean the filter cloth 008 and filter element 009. Open the second material valve 102 to clean the pre-coating reflux pipeline. When the preset cleaning time is up, open the filter forward air inlet valve 403 to discharge the wastewater to the sewage station;

[0113] 9.3.5 Cleaning of the filter liquid tank 012: Open the cleaning branch valve 203, cleaning branch valve 204, filter liquid tank ball valve 209, filter liquid tank bottom valve 109, filter liquid tank drain valve 705 to clean the inside of the filter liquid tank and the discharge pipeline. When the preset cleaning time is up, close the filter liquid tank drain valve 705, open the filter liquid delivery pump 020, filter liquid connecting valve 110, filter liquid sewage main valve 708, filter liquid drainage valve 710, and wastewater main valve 712 to clean the filter liquid delivery pipeline, discharge the wastewater to the sewage station, empty the pump, and open the pump back sewage valve 707 to discharge the residual liquid.

[0114] The above steps are performed in sequence according to the cleaning time and step sequence settings.

[0115] 9.4 Verify the cleaning effect: Open the water inlet valves in sequence through all pipes and tanks, and complete the cleaning procedure after the conductivity meter 022 detects and passes.

[0116] Step 10: Steam-in-place sterilization (SIP).

[0117] 10.1 Preparation before sterilization: Open the main steam valve 301, and open the steam trap 702, steam trap 706, and steam trap 709;

[0118] 10.2 Heating: Open cleaning branch valve 203, cleaning branch valve 204, cleaning branch valve 205, upper cleaning ball valve 206, lower cleaning ball valve 207, cleaning branch valve 208, filtered liquid tank cleaning ball valve 209, cleaning branch valve 214, cleaning branch valve 216, cleaning branch valve 217, open the feed tank bottom valve 103 and material connecting valve 104, and heat the entire system to 90°C.

[0119] 10.3 Sterilization first stage: Open the exhaust overflow valve 113 and the exhaust overflow valve 2 114 to sterilize the exhaust overflow pipeline, open the cleaning branch valve 214, the cleaning branch valve 216, the cleaning branch valve 217 and the filter feed valve 105 to sterilize the pipelines before and after the feed pump, open the tank bottom valve 103, the material connecting valve 104, and the tail valve 117 to sterilize the material pipeline, the tail pipeline and the return pipeline, and open the filtrate drainage valve 708, valve 211, the filtrate tank washing ball valve 209, and valve 210 to sterilize the alkali liquid circulation pipe. The sterilization temperature is 121 to 123°C, which is measured and controlled by the pipeline end temperature sensor TCR, and the insulation time is 30 minutes.

[0120] 10.4 Second stage of sterilization: Open the cleaning branch valve 203, cleaning branch valve 204, cleaning branch valve 205, and upper cleaning ball valve 206 to sterilize the feed tank, open the cleaning branch valve 208, and the filter tank cleaning ball valve 209 to sterilize the filter tank, open the cleaning branch valve 212, the pre-coating reflux valve 115, the filter reverse air inlet valve 402, and the filter forward air inlet valve 403 to sterilize the filter forward air pipe, open the cleaning branch valve 214, valve 215, valve 118, filter feed valve 105, and return valve 116 to sterilize the filter return pipe. The sterilization temperature is 121 to 123°C, which is measured and controlled by the pipe end temperature sensor TCR, and the insulation time is 30 minutes.

[0121] 10.5 Cooling down the first stage: When the sterilization setting time is reached, close the steam main valve 301, start cooling the feed tank and pipeline, open the steam trap 702 to cool the system, and TCR002 to judge the temperature. When cooling the filter and pipeline, open the steam trap 703 to cool the system, and TCR003 to judge the temperature. When cooling the clear liquid tank and pipeline, open the steam trap 706 and steam trap 709 to cool the system, and TCR004 to judge the temperature.

[0122] 10.6 Maintaining pressure: During the cooling process, the entire system maintains a slightly positive pressure state. If necessary, open the filter's forward air inlet valve 403 to maintain pressure.

[0123] Figure 4 The solid-liquid separation method of pneumococcal fermentation lysate in the prior art has the following technical problems and defects: the process is relatively complex, the production links are many, the cost of using the deep filter is high, the cleaning in place (CIP) and sterilization in place (SIP) processes are cumbersome, and the batch production cycle is long; it is impossible to achieve complete batch solid-liquid separation, and there is a situation where tailings are forced to be discarded, the yield is low, and the cost is increased; the fluidized slag material is discharged, the volume of inactivated materials is large, the cost is high, and the cost of sewage treatment is high; the filter material of the deep filter is a disposable filter device, the replacement process requires manual intervention, and the cost of use is high; the continuous flow centrifuge must be maintained regularly (more than once a year), the maintenance cost is high, the maintenance work is complicated, and the maintenance technology requirements are high; the continuous flow centrifuge operates with high-speed rotating moving parts, which consumes a lot of power, has certain risks in operation, and has high operating noise, which is not environmentally friendly.

[0124] Figure 1 The novel cake layer candle solid-liquid separation system for biopharmaceutical production used in Example 1 is shown. Figure 1 As shown, the cake layer candle solid-liquid separation system for biopharmaceutical grade is composed of a solid-liquid separation system, a formulated filter aid that has been screened through experiments, and a PLC control program.

[0125] Figure 2The figure shows the filter cake layer candle type solid-liquid separation system. Figure 3 The working state diagram of the filter element assembly in the cake layer candle type solid-liquid separation system is shown in FIG. 1 , where the filter cloth 008, the filter element supporting orifice plate layer 091, the filter element central tube wall 092, and the filter cake layer 010 are located in the filter element 009. Figure 3 shown.

[0126] Figure 5 The flowchart of the method in Example 1 is shown. The present invention uses a novel cake layer candle solid-liquid separation system to replace the continuous flow disc separator + deep filter process method combination in the existing process method, shortens the process production process, simplifies the operation process and production system, improves the product yield, and achieves energy saving and consumption reduction, reduces production costs, and is environmentally friendly.

[0127] Table 1 is a comparison table of the cake layer candle type solid-liquid separation system in Example 1 of the present invention and the "continuous flow disc separator + deep filter combination" in the prior art.

[0128] Table 1:

[0129]

[0130] Figure 6 The figure shows the effect of solid-liquid separation of the inactivated pneumococcal fermentation broth, wherein the left side is a photo of the clarified liquid after solid-liquid separation by the continuous flow disc centrifuge in the prior art, with a turbidity of NTU30-100; the right side is a photo of the clarified liquid obtained after solid-liquid separation by the cake layer candle-type solid-liquid separation system in Example 1 of the present invention, with a turbidity of ≤10NTU, and the NTU value of the material in the photo on the right is 3NTU.

[0131] In order to meet the high sanitation requirements of biopharmaceutical-grade production equipment and the requirements of no dead corners and easy cleaning, the core equipment of the cake layer candle solid-liquid separation system provided by the present invention, the cake layer candle filter equipment structure has been significantly optimized and upgraded:

[0132] Traditional candle filter cartridges have many dead corners for cleaning, which cannot meet the requirements of high-level sanitary biopharmaceutical equipment with no dead corners and easy cleaning. The preferred core optimization design content of the present invention is: redesigning a special integrally formed filter cartridge that can be used in biopharmaceutical production, which is recorded in the Chinese utility model patent ZL202223038887.2, the patent application number: CN202223038887.2, application date: November 11, 2022; publication number: CN218901009U, publication date: April 25, 2023.

[0133] The design, material and polishing grade of all pipelines in contact with materials, cleaning fluid and injection-grade water in the cake layer candle solid-liquid separation system of the present invention are optimized, and the dead zone of all pipelines shall not be greater than 2-3 times the inner diameter (2D, 3D principle), so as to fully meet the high-level technical requirements of no dead angle and easy cleaning. For the parts that cannot meet the 2D and 3D principles due to hardware specification limitations, adjustments are made in the control program to meet the cleaning requirements.

[0134] Example 2

[0135] Example 2 provides preferred filter aid types and formulations of the present invention, which are applied in step 2 of Example 1.

[0136] Embodiment 2 Preferred filter aids are:

[0137] The preferred brand of diatomaceous earth is Celpure, and the preferred model is at least one of Celpure C25, Celpure C65, Celpure C100, Celpure C300, and Celpure C1000.

[0138] The preferred brand of cellulose is Riedenmeier, and the preferred model is at least one of the ARBOCEL and / or VIVAPUR series.

[0139] The preferred brand of silicon dioxide is Longhua Chengxin Powder Technology Co., Ltd., and the preferred model is at least one of LH102 and LH104.

[0140] On this basis, we further selected 1 # Diatomaceous earth, 2 # Cellulose 3 # Silicon dioxide powder.

[0141] 1 # Diatomaceous earth (Celpure C1000: Celpure C65=8:2) has the advantages of better filtration accuracy, faster initial filtration rate, faster filtration rate decay, and larger addition amount.

[0142] 2 # Cellulose (at least one of ARBOCEL15, 16, 17, 18) has poor filtration accuracy; fast initial filtration rate, slow filtration rate decay; large addition amount.

[0143] 3 # Silica powder (LH102) has good filtration accuracy; the initial filtration rate is slow, the filtration rate decays quickly; and the addition amount is small.

[0144] Furthermore, through many experimental comparisons and data analysis, the formula of the composite filter aid and the applicable price are selected as follows:

[0145] Recipe 1: 1 # Diatomaceous earth: 2 # Fiber: 3 # Silica powder = 6:2:1 weight ratio. Applicable price types: 1, 2, 6A, 7F, 9N, 9V, 10A, 11A, 12F, 17F, 19A, 19F, 33F.

[0146] Recipe 2: 1 # Diatomaceous earth: 2 # Fiber: 3 # Silica powder = 5.5:2:1.5 weight ratio. Applicable price types: 4, 5, 22F, 23F.

[0147] Recipe 3: 1 # Diatomaceous earth: 2 # Fiber: 3 # Silicon dioxide powder = 6:3:1 weight ratio. Applicable price types: 14, 15B, 18C, 20.

[0148] Recipe 4: 1 # Diatomaceous earth: 2 # Fiber: 3 # Silica powder = 7:2:1 weight ratio. Applicable valence types: 3, 8.

[0149] Example 3

[0150] Exploration of the formula of compound filter aid:

[0151] The new solid-liquid separation method of the present invention needs to be applicable to 23 common pneumococcal valence types (1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F, 33F).

[0152] In actual use, the material properties of the 23 types of pneumococci are inconsistent, and even the material properties of fermentation products of the same valence type in different batches are very poorly consistent, resulting in very large individual differences in solid-liquid separation. When a filter aid formula that is very ideal for filtering a certain valence type is applied to another valence type, it may not even be possible to complete the filtration. Therefore, it is necessary to explore the best ratio of filter aid formula for solid-liquid separation of each valence type material according to the different characteristics of each of the 23 types of pneumococci. This is also a very critical process step and process for the cake layer candle solid-liquid separation system to achieve good solid-liquid separation effect in pneumococcal lysate.

[0153] Due to the diversity and variability of the processed materials, many factors that affect the solid-liquid separation effect cannot be predicted, and it is impossible to effectively guide the filter aid formulation through conventional theoretical methods. The only way to obtain a more ideal solid-liquid separation effect is to explore a suitable filter aid formulation through repeated experiments.

[0154] Filter aid formula:

[0155] With diatomaceous earth, cellulose and silicon dioxide as the main components of the filter aid formula, the proportions and models of various filter aids were adjusted respectively. Data were obtained through actual material filtration tests and the filtration clarity and filtration speed were compared to select the best filter aid formula.

[0156] Because there are too many types of 23 pneumococci in actual production, different filter aid formulas need to be used when converting materials of different valence types, which is cumbersome and prone to use errors in actual production applications. Therefore, after optimizing the best filter aid formulas of various valence types, the differences in filter aid formulas suitable for different materials are compared, and the most suitable filter aid formula that can be applied to several different materials is selected, so as to reduce the types of filter aid formulas and facilitate use.

[0157] In Example 3, the operating steps for exploring the filter aid formulation are as follows:

[0158] 1. Observe and partially understand the material conditions of pneumococcal fermentation lysate, mainly the fluidity, viscosity, solid content and other indicators, and preliminarily determine the type and proportion of filter aid to be added and the filter aid coating process based on intuitive impression.

[0159] 2. Add the selected filter aid to the test material in a predetermined proportion, adjust the volume to 200ml, and stir evenly.

[0160] 3. Use the filter to test the filterability, put the prepared filter aid material into the filter, supply clean compressed air, and measure the filtration rate in the filtration characteristics. At the same time, continuously record the clear liquid time and the volume of the clarified liquid.

[0161] 4. When the filtration rate drops to an unacceptable rate (needs to be determined comprehensively as appropriate) or all the raw materials have been filtered, stop filtering and record.

[0162] 5. Analyze the above process, record the data into statistics to form charts, and perform data analysis.

[0163] 6. According to the above analysis results and the test process, make or adjust the proportion of filter aids, or change the type of filter aids, or change the coating form of filter aids and other subsequent work content adjustments, so as to increase the filtration speed, improve the clarity of the clear liquid, reduce the proportion of filter aids added, and reduce the cost of using filter aids.

[0164] 7. Repeat the above 2-6 processes until preliminary satisfactory and conclusive test results are obtained.

[0165] 8. According to the test conclusions obtained from the filterability qualitative test, prepare the test materials and adjust the volume to 2-5L.

[0166] 9. Use the filtration characteristics to test the filtration system. Inject the prepared material into the filter feed tank, start the pneumatic diaphragm pump, and pump the material into the filter until the overflow reflux has material reflux.

[0167] 10. Pre-coating of filter aid.

[0168] 11. When the material in the pre-coating reflux pipe is clear, stop pre-coating and collect and filter the clarified liquid.

[0169] 12. Stop feeding when the material in the feed tank is almost used up, and stop filtering when there is no clear liquid discharged from the clear liquid pipe.

[0170] 13. Use compressed air to blow dry the filter cake in a forward direction to further collect the valuable substances in the filter cake.

[0171] 14. Blow compressed air in the reverse direction to blow off the filter cake and complete the cake unloading.

[0172] 15. Collect the filter cake that has fallen off the filter element, weigh it and measure the thickness, bulk density, etc. of the filter cake.

[0173] 16. Analyze the obtained filtration characteristic test results and compare them with the test data recorded in the filterability qualitative test to obtain the filtration characteristic data of the tested material and the characteristic data of the clarified liquid and the filter cake.

[0174] 17. Based on the test data results of the filtration system for filtration characteristics testing, analyze the possible results obtained using the cake layer candle solid-liquid separation system, and perform theoretical calculations and comparisons of various technical and economic indicators as a basic support data system for production verification testing.

[0175] 18. Use the cake layer candle solid-liquid separation system full function verification system for testing.

[0176] 19. Prepare and adjust the volume to 80-100L of the material to be tested.

[0177] 20. The first few steps of the test process are the same as steps 9-11 above.

[0178] 21. After the material in the feed tank is processed, the remaining material is processed. At this time, the remaining material in the filter enters the filter through the top and passes through the filter cake layer together with the compressed air.

[0179] 22. After the remaining materials in the filter are processed, compressed air is introduced to dry the formed filter cake and further recover the valuable substances in the filter cake.

[0180] 23. After the filter cake is dried, if there is a process requirement for further drying of the filter cake, heated compressed air can be introduced for drying. If there is no such requirement, this step can be skipped.

[0181] 24. Perform the slag discharge operation and introduce compressed air in the reverse direction to discharge the cake. The discharged filter cake is discharged into the closed slag discharge mechanism.

[0182] 25. After the slag discharge operation is completed, close the slag discharge bottom valve and tighten the slag bag.

[0183] 26. Open the locking device, take out and remove the slag bag.

[0184] 27. Put the slag into the bag, restore the closed slag discharge mechanism and lock it.

[0185] 28. The collected clarified liquid is sent out for subsequent processing.

[0186] Operation steps 20-24 are controlled by PLC program and are fully automatic.

[0187] At this point, the full functional test verification of pneumococcal lysate has been completed.

[0188] The specific operating steps and program operation process control points of various price types are not completely consistent. This is determined by the different material characteristics. Recipe menu management is done in the PLC control program and selected according to the material being processed during use.

[0189] 29. The test results indicate that among the 1, 2, 3, 4, 5, 6A, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F, and 33F price types of pneumococcus, according to the filtration difficulty and the clarity that can be achieved to meet the ultrafiltration feed requirements of the membrane package, they can basically be divided into 4 types of 4 groups of filter aid formulations, locking the formulation menu categories.

[0190] The screening in Example 3 yielded the preferred scheme for the types and formulations of the filter aids described in Example 2.

[0191] Example 4

[0192] Screening of filter aid formulations for clarification and filtration of 23-valent Streptococcus pneumoniae fermentation lysate.

[0193] Through orthogonal experiments, the filter aid formula used in the clarification and filtration process of multivalent Streptococcus pneumoniae fermentation lysate was optimized and screened.

[0194] The screening principle of Example 4 is:

[0195] 1. The clarification and filtration process of polyvalent pneumococcal fermentation lysate is affected by many factors. The results of the preliminary basic experiments (using a single type of filter aid for filtration experiments) show that the type, specification and addition amount of the filter aid are the main influencing factors, and there are many problems with the use of a single type of filter aid, including the filtration accuracy not meeting the requirements, the filtration speed is too slow, the amount of filter aid added is too large, etc. The use of a combination of multiple filter aids is very likely to improve the filtration effect.

[0196] 2. Performance of alternative filter aids:

[0197] a) 1 # Diatomaceous earth (C1000:C65=8:2)——better filtration accuracy; faster initial filtration rate, faster filtration rate decay; larger addition amount.

[0198] b) 2 # Cellulose (ARBOCEL15-18) - poor filtration accuracy; fast initial filtration rate, slow filtration rate decay; large addition amount.

[0199] c) 3 # Silica powder (LH102) - good filtration accuracy; slow initial filtration rate, fast filtration rate decay; small addition amount.

[0200] 3. Use filter aids with different ratios and add 2% of the mass of the feed liquid (the upper limit of the amount of filter aid added that the user wants), conduct filtration experiments, record the filtration flow rate, and measure the turbidity of the filtrate.

[0201] 4. Select the optimal formula based on the filtration flow rate and filtrate turbidity.

[0202] The specific steps of screening in Example 4 are:

[0203] Step a: Prepare the test materials and adjust the volume of the raw material liquid to 2L.

[0204] Step b: Use the filtration characteristics to test the filtration system, inject the configured material into the filter feed tank, start the pneumatic diaphragm pump, and pump the material into the filter until there is material reflux in the overflow reflux.

[0205] Step c: pre-coating of filter aid, the amount of filter aid added is 2% of the mass of the raw material liquid.

[0206] Step d: introducing the raw material solution and collecting the filtered clarified liquid.

[0207] Step e: Stop feeding when the material in the feed tank is nearly used up, and stop filtering when no clear liquid is discharged from the clear liquid pipe.

[0208] Step f: Use compressed air to blow dry the filter cake in a forward direction to further collect the valuable substances in the filter cake.

[0209] Step g: Blow compressed air in the reverse direction to blow off the filter cake and complete the cake unloading.

[0210] Step h: Collect the filter cake that has fallen off the filter element, weigh it and measure the thickness, bulk density, etc. of the filter cake.

[0211] Step i: Analyze the filtration characteristic data of the tested material and the characteristic data of the clarified liquid and the filter cake.

[0212] The following is the orthogonal experimental table design and experimental results analysis in Example 4, where Table 2 is the flow rate detection results, Table 3 is the turbidity,

[0213] Table 2: Flow rate (L / m 2 / h)

[0214]

[0215] In Table 2, according to the analysis of the experimental results, B has the largest range, so it has the greatest impact on the experimental results, followed by A, and C is the smallest. Then analyze the mean value, and finally determine that the condition for obtaining the maximum flow rate is A2B3C1.

[0216] Table 3: Turbidity (NTU)

[0217]

[0218] According to the experimental results recorded in Table 3, A has the largest range, so it has the greatest impact on the experimental results, followed by B and C. Then analyze the mean and finally determine that the best condition for obtaining turbidity is A1B1C3.

[0219] The best solution selection principle in Example 4 is:

[0220] a) The mean of the extreme difference is the largest, which has a great impact on the experimental results;

[0221] b) For the main factors, select the level that best indicates the condition;

[0222] c) For minor factors, select the average level based on the principle of economy and convenience.

[0223] The best solution determined on this basis is:

[0224] d) 1 #The addition ratio of diatomaceous earth is between 5-7, choose 6;

[0225] e) 2 # The cellulose addition ratio is between 1-3, choose 2;

[0226] f) 3 # The addition ratio of silicon dioxide powder is between 0.5-1.5, select 1.

[0227] According to the experimental results of Example 4, the preferred formula of the composite filter aid of the present invention is: 1 # Diatomaceous earth: 2 # Fiber: 3 # Silicon dioxide powder = 6:2:1. Suitable for solid-liquid separation of 1, 2, 6A, 7F, 9N, 9V, 10A, 11A, 12F, 17F, 19A, 19F, 33F valence types of pneumococcus.

[0228] If other preferred formulas are needed, further screening experiments can be carried out based on the methods and results of this experiment.

[0229] The above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A solid-liquid separation method for 23-valent pneumococcal fermentation lysate, comprising the following steps: Step 1: Select the filter aid formula: select the corresponding formula according to the valence type of the pneumonia vaccine fermentation lysate to be processed; Step 2: Prepare a filter aid, and evenly mix the formulated filter aid with the fermentation lysate of the pneumonia vaccine to be processed in a feed tank; the formulated filter aid and the solids to be removed in the fermentation lysate of the pneumonia vaccine to be processed form large solid particles in a cross-linked state; The recipe described in step 1 includes 1 # Diatomaceous earth, 2 # Fiber and 3 # Silicon dioxide powder, 1 # The diatomaceous earth is composed of Celpure C1000 and Celpure C65 diatomaceous earth of Yiruishi, and the mass ratio is Celpure C1000: Celpure C65=8:2; 2 # The cellulose is at least one of the celluloses ARBOCEL15-18 of the ARBOCEL series produced by Ruidenmeier; 3 # The silicon dioxide powder is the silicon dioxide powder of model LH102 produced by Longhua Chengxin Powder Technology Co., Ltd.; The filter aid formulation is: Recipe 1: 1 # Diatomaceous earth: 2 # Fiber: 3 # The mass ratio of silicon dioxide powder is 6:2:1, which is suitable for solid-liquid separation of 1, 2, 6A, 7F, 9N, 9V, 10A, 11A, 12F, 17F, 19A, 19F, and 33F valence types of pneumococci; Step 3: Filter the mixed material in step 2 through a sanitary filter cake layer filter, which contains a filter cloth and a filter element; after filtration, the filtrate flows back to the feed tank, and the large solid particles in the cross-linked state contained in the mixed material in step 2 are intercepted by the filter cloth, forming a filter cake layer on the surface of the filter cloth; the filter cake layer constructs a flow channel due to the cross-linking bridging effect of the solid particles, allowing liquid and small solid particles to pass through; and as the filter cake layer thickens, the particle size of the solid particles allowed to pass through the filter cake layer flow channel becomes smaller, until the turbidity of the filtrate is ≤10NUT; Step 4: stop the filtrate from flowing back to the feed tank, transfer the filtrate to the filtrate tank, and obtain filtrate; Step 5: Treatment of residual tailings: The remaining materials in the sanitary filter cake layer filter are controlled by air pressure to intercept the solids into the filter cake layer; Step 6: washing the filter cake; Step 7: Filter cake unloading: let the filter cake layer on the filter cloth fall off into the closed slag discharge mechanism; Step 8: Transfer of filtrate: Transfer all filtrate to the ultrafiltration department.

2. The solid-liquid separation method of 23-valent pneumococcal fermentation lysate according to claim 1, characterized in that: The filter aid formulation is: Recipe 2: 1 # Diatomaceous earth: 2 # Fiber: 3 # The mass ratio of silica powder is 5.5:2:1.5, which is suitable for solid-liquid separation of 4, 5, 22F, and 23F valence types of pneumococci; Recipe 3: 1 # Diatomaceous earth: 2 # Fiber: 3 # The mass ratio of silica powder is 6:3:1, which is suitable for solid-liquid separation of pneumococcal 14, 15B, 18C, and 20 valent forms; Recipe 4: 1 # Diatomaceous earth: 2 # Fiber: 3 # The mass ratio of silicon dioxide powder is 7:2:1; it is suitable for solid-liquid separation of 3- and 8-valent forms of pneumococcus.

3. The solid-liquid separation method of 23-valent pneumococcal fermentation lysate according to claim 2, characterized in that: The specific steps of step 2 are: Step 2.1: inject the fermentation liquid to be treated into the feed tank and start stirring; Step 2.2: Weigh and prepare the filter aid according to the formula, add the prepared filter aid into the feed tank under bacteria-controlled and closed conditions, and mix it evenly with the fermentation lysate of the pneumonia vaccine to be treated in the tank under stirring.

4. The solid-liquid separation method of 23-valent pneumococcal fermentation lysate according to claim 3, characterized in that: The specific steps of step 3 are: Step 3.1: Feeding and filling: conveying the mixed material in step 2 to a sanitary filter cake layer filter; the filter element is a sanitary perforated plum blossom filter element; Step 3.2: pre-coating filtration, filter cake layer is formed, and the filtrate reaches the process index: the material to be treated is filtered through the filter cloth and the filter element, and the filtrate is returned to the feed tank; The large solid particles in a cross-linked state formed in the material to be treated are intercepted by the filter cloth, forming a filter cake layer on the surface of the filter cloth. The flow channel constructed by the cross-linking bridging effect of the solid particles in the filter cake layer allows liquid and some small solid particles to pass through; under the action of a pump pressure of 0.3 to 0.35MPa, the filter cloth is pressed tightly against the filter element, and the cross-section is plum blossom-shaped.

5. The solid-liquid separation method of 23-valent pneumococcal fermentation lysate according to claim 4, characterized in that: In step 4, when the filtrate standard meets the turbidity requirement of the filtration target, the filtrate is controlled by a valve to enter the filtrate tank to obtain the filtrate; Step 5: Treatment of residual tailings: The sanitary cake layer filter is provided with a distributor, and the residual material is sprayed out from the distributor. The air is compressed in the sanitary cake layer filter to form a gas-liquid-solid mixture. The pressure in the sanitary cake layer filter is controlled to be 0.3 to 0.35 MPa, so that the gas-liquid-solid mixture passes through the cake layer and the filter cloth under the action of pressure, the solid matter is intercepted by the cake layer, the liquid enters the filter liquid tank, and the gas is discharged into the exhaust system through the filter liquid tank exhaust valve; this process continues until all materials in the sanitary cake layer filter are completely processed.

6. The solid-liquid separation method of 23-valent pneumococcal fermentation lysate according to claim 5, characterized in that: The specific steps of step 6 are: Step 6.1: Fill the feed tank with water at a temperature of 20°C. Inject water to preliminarily wash the feed tank, and use the wash water to wash the filter cake; Step 6.2: Washing filter cake: The washing water in the feed tank is transported to the sanitary filter cake layer filter to dilute the previous residual material in the filter. After the feed tank is emptied, repeat step 5 to treat the remaining tailings, and spray the filter cake with washing water until all the washing water in the sanitary filter cake layer filter enters the filtrate tank.

7. The solid-liquid separation method of 23-valent pneumococcal fermentation lysate according to claim 6, characterized in that: In step 7, a closed slag discharge mechanism is connected below the sanitary filter cake layer filter. By controlling the direction of the airflow in the sanitary filter cake layer filter, compressed air is used to act on the inside of the filter cloth and the filter element from downstream to upstream. The filter cloth with a plum blossom-shaped cross-section is expanded into a circular shape under pressure, so that cracks appear in the plum blossom depression on the filter cloth, and the filter cake falls off the filter cloth and falls to the closed slag discharge mechanism.

8. The solid-liquid separation method of 23-valent pneumococcal fermentation lysate according to claim 7, characterized in that: In step 8, after the filter cake washing in step 6.2 is completed, the filtrate is transferred to the ultrafiltration department.

9. The solid-liquid separation method of 23-valent pneumococcal fermentation lysate according to claim 8, characterized in that: It also includes a CIP cleaning step and a SIP steam sterilization step.

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