CN114618304B - Ultrafiltration purification system for porcine reproductive and respiratory syndrome virus - Google Patents

Ultrafiltration purification system for porcine reproductive and respiratory syndrome virus Download PDF

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Publication number
CN114618304B
CN114618304B CN202210327192.1A CN202210327192A CN114618304B CN 114618304 B CN114618304 B CN 114618304B CN 202210327192 A CN202210327192 A CN 202210327192A CN 114618304 B CN114618304 B CN 114618304B
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ultrafiltration membrane
stirring
ultrafiltration
concentration tank
purification system
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CN114618304A (en
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李阳
沈建军
彭俊平
胡永明
娄坤鹏
张秀文
陈雪
屈建
王悦芸
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Zhejiang Mibolerone Biological Technology Co ltd
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
    • B01D61/14—Ultrafiltration; Microfiltration
    • B01D61/145—Ultrafiltration
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
    • B01D61/14—Ultrafiltration; Microfiltration
    • B01D61/18—Apparatus therefor
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D67/00—Processes specially adapted for manufacturing semi-permeable membranes for separation processes or apparatus
    • B01D67/0079—Manufacture of membranes comprising organic and inorganic components
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D69/00—Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor
    • B01D69/02—Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor characterised by their properties
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D71/00—Semi-permeable membranes for separation processes or apparatus characterised by the material; Manufacturing processes specially adapted therefor
    • B01D71/06—Organic material
    • B01D71/66—Polymers having sulfur in the main chain, with or without nitrogen, oxygen or carbon only
    • B01D71/68—Polysulfones; Polyethersulfones
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/02—Separating microorganisms from the culture medium; Concentration of biomass
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D2325/00—Details relating to properties of membranes
    • B01D2325/24—Mechanical properties, e.g. strength
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D2325/00—Details relating to properties of membranes
    • B01D2325/36—Hydrophilic membranes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/10011—Arteriviridae
    • C12N2770/10051—Methods of production or purification of viral material

Abstract

The invention relates to the technical field of virus propagation, in particular to an ultrafiltration purification system for porcine reproductive and respiratory syndrome virus, which comprises a first concentration tank, a second concentration tank and a third concentration tank which are sequentially connected through pipelines, wherein a micro-filter is arranged in front of the first concentration tank, a first ultrafiltration membrane package is arranged between the first concentration tank and the second concentration tank, a second ultrafiltration membrane package is arranged between the second concentration tank and the third concentration tank, a first ultrafiltration membrane and a second ultrafiltration membrane are arranged in the first ultrafiltration membrane package and the second ultrafiltration membrane package Bie An, the aperture of the first ultrafiltration membrane is larger than the molecular weight of target virus, and the aperture of the second ultrafiltration membrane is smaller than the molecular weight of target virus. According to the ultrafiltration purification system for the porcine reproductive and respiratory syndrome virus, provided by the invention, through the three-stage filtration purification system, more than 95% of invalid proteins in the vaccine can be removed, the antigen purity can be effectively improved, and the stress response of animals to the vaccine can be reduced.

Description

Ultrafiltration purification system for porcine reproductive and respiratory syndrome virus
Technical Field
The invention relates to the technical field of virus propagation, in particular to an ultrafiltration purification system for porcine reproductive and respiratory syndrome virus.
Background
Porcine reproductive and respiratory syndrome (porcine reproductive and respiratory syndrome virus, PRRS) is a virulent contagious disease, and the outbreak disease is characterized by abortion (10% -50%), death of sows (5% -10%), death of commercial pigs exceeding 50%, death of piglets before weaning and respiratory diseases, and in recent years, the disease presents a remarkable high-rise trend in China, causes great loss to the pig industry and becomes one of important infectious diseases seriously threatening the development of the Chinese pig industry.
The pathogen of PRRS is Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), which is an RNA virus with high mutation rate discovered so far, and the PRRSV can be horizontally transmitted and vertically transmitted. The pigs are infected by various ways such as oral cavity, nasal cavity, muscle, abdominal cavity, genital tract and the like, and the infected pigs expel toxin from saliva, urine, semen and milk. After PRRSV invades the pig body, the immune system is destroyed, so that the immunity of the body is reduced, and the PRRSV can be mixed with the infection of the type II circovirus, the parvovirus disease, the pseudorabies virus disease and the like.
The prior main technical means for preventing and treating the disease is PRRS vaccine, and the porcine reproductive and respiratory syndrome inactivated vaccine produced by the conventional process has a certain side reaction in clinical application, and is mainly related to the external antigen component (ineffective protein) contained in the virus antigen. Such as culture medium, serum and host cell debris proteins in the production process, which can cause allergic reaction of organisms and possibly cause the organisms to produce antibodies to protein drugs, the purity of the antigen must be considered while the concentration of the antigen is emphasized, the higher the purity is, the lower the impurity protein content is, and the less the animal is stressed to the vaccine.
Disclosure of Invention
Therefore, the invention aims to provide an ultrafiltration purification system for porcine reproductive and respiratory syndrome virus, which can remove more than 95% of invalid proteins in a vaccine by the aid of the three-stage filtration purification system, can effectively improve the antigen purity and reduce the stress response of animals to the vaccine.
The invention solves the technical problems by the following technical means:
the utility model provides a pig breeds and breathes syndrome virus ultrafiltration purification system, includes first concentrated jar, second concentrated jar and the third concentrated jar that connects gradually through the pipeline, be provided with the microfilter before the first concentrated jar, be provided with first milipore filter package between first concentrated jar and the second concentrated jar, be provided with the second milipore filter package between second concentrated jar and the third concentrated jar, bie An are equipped with first milipore filter, second milipore filter in first milipore filter package and the second milipore filter package, the aperture of first milipore filter is greater than the molecular weight of target virus, the aperture of second milipore filter is less than the molecular weight of target virus.
Further, the pore diameter of the first ultrafiltration membrane is 35-40nm, and the pore diameter of the second ultrafiltration membrane is 15-25nm.
Further, the first ultrafiltration membrane and the second ultrafiltration membrane are both modified polyethersulfone ultrafiltration membranes, and the modified polyethersulfone ultrafiltration membranes comprise the following raw materials in parts by weight: 95-100 parts of polyethersulfone, 3-5 parts of polyvinylpyrrolidone, 15-20 parts of composite modified particles and 2-4 parts of dimethylacetamide.
Further, the composite modified particles are formed by modifying modified montmorillonite through polyacrylic acid intercalation and then compositing the modified montmorillonite with nano cellulose crystals.
According to the composite modified particles, the montmorillonite belongs to a multi-layer structure, the modified montmorillonite is subjected to intercalation modification through the polyacrylic acid, so that the distance between the modified montmorillonite layers can be increased, the modified montmorillonite is further subjected to layer stripping, and is added into a matrix material, so that the sliding performance of the matrix material is improved, film forming is facilitated, meanwhile, the tensile strength of an ultrafiltration film is improved, meanwhile, the added polyacrylic acid and polyethersulfone have good compatibility, the two can also undergo a crosslinking reaction, the compatibility between the composite modified particles and the matrix material can be improved through the addition of the polyacrylic acid, the interface effect between the composite modified particles and the matrix material is weakened, and the modification effect of the composite modified particles on the matrix material is better; on the one hand, the nano-cellulose crystal has good hydrophilicity, so that the hydrophilicity of the ultrafiltration membrane can be effectively improved, the pollution resistance of the ultrafiltration membrane is improved, on the other hand, the nano-cellulose crystal is not easy to agglomerate in a matrix material due to the fact that the modified montmorillonite plays a certain constraint role on the nano-cellulose crystal, the nano-cellulose crystal can be well dispersed in the matrix material, and the membrane modification effect is better.
Further, the preparation method of the modified polyethersulfone ultrafiltration membrane comprises the following steps: adding the pretreated composite modified particles into dimethylacetamide, performing ultrasonic dispersion for 30-45min, adding polyether sulfone, stirring and dissolving, adding ammonium persulfate and sodium dodecyl benzoate, continuously introducing nitrogen for 30min, heating to 80-95 ℃ under nitrogen atmosphere, performing heat preservation and stirring reaction for 3-5h, then adding polyvinylpyrrolidone, continuously stirring and reacting for 2-3h to obtain a casting solution, stopping stirring, performing heat preservation and standing and defoaming for 6h, cooling to room temperature, scraping a film, performing cooling and molding, then soaking in distilled water for 2-3d, and drying to obtain the modified polyether sulfone ultrafiltration membrane.
Further, the pretreatment of the composite modified particles is as follows: stirring and dispersing the composite modified particles in deionized water, adding ammonium ceric nitrate and concentrated sulfuric acid, continuously stirring at normal temperature for reaction for 1-2h, filtering after the reaction is finished, and washing a filter cake to be neutral and drying.
The composite modified particles are subjected to pre-oxidation treatment by ceric ammonium nitrate and concentrated sulfuric acid, so that the combination between the composite modified particles and polyethersulfone can be facilitated.
Further, the preparation method of the composite modified particles comprises the following steps:
s1: stirring and dispersing the modified montmorillonite in an acetone solution, adding an acrylic acid monomer, stirring and mixing uniformly by magnetic force, adding hexadecyl trimethyl ammonium bromide and benzoyl peroxide amine, stirring and mixing uniformly, heating to 100-120 ℃ in an oil bath, reacting for 90-120min at a constant temperature, and then removing acetone in vacuum at room temperature to obtain a material I;
s2: dispersing nanocellulose crystal in toluene by stirring, ultrasonically dispersing to obtain a suspension, placing the mixed material I in a ball mill, adding the suspension, ball milling for 30-45min at the speed of 200-300rpm, taking out, decompressing, recovering toluene, drying to obtain solid, stirring in the heating process of far infrared heating to 70-75 ℃, simultaneously spraying 1-2wt% chitosan acetic acid solution, continuing stirring for 10-15min after spraying, and drying to obtain composite modified particles.
In the preparation process of the composite modified particles, ball milling is utilized to improve the surface energy of the material I and the nano cellulose crystals, so that the material I and the nano cellulose crystals are promoted to be compounded, meanwhile, the ball milling is carried out, then chitosan solution is sprayed, under the condition of infrared heating, chitosan is separated out, the nano cellulose crystals and the modified montmorillonite can be more stably compounded together, and meanwhile, the chitosan also has good hydrophilicity, so that the enhancement of the pollution resistance of the composite modified particles to polyethersulfone is facilitated.
Further, the preparation method of the modified montmorillonite comprises the following steps: stirring and dispersing montmorillonite in 0.9wt% physiological saline, heating to boiling, cooling to room temperature, filtering and washing, stirring and dispersing in deionized water, dripping tetramethylammonium chloride solution, reacting for 4 hours at 80-90 ℃, and carrying out suction filtration, washing, drying and grinding after the reaction is completed to obtain the modified montmorillonite.
The montmorillonite is soaked in normal saline to increase the interlayer distance of the montmorillonite, and then activated by the tetramethyl ammonium chloride solution, which is more beneficial to subsequent reaction
Further, in the preparation of the modified montmorillonite, the mass ratio of the montmorillonite to the tetramethyl ammonium chloride solution is 1 (0.2-0.5).
The invention has the beneficial effects that:
1. according to the ultrafiltration purification system for the porcine reproductive and respiratory syndrome virus, provided by the invention, through the three-stage filtration purification system, more than 95% of invalid proteins in the vaccine can be removed, the antigen purity can be effectively improved, and the stress response of animals to the vaccine can be reduced.
2. The ultrafiltration purification system for the porcine reproductive and respiratory syndrome virus uses the modified polyethersulfone ultrafiltration membrane for filtration, has good anti-pollution performance, and can reduce the blockage of virus proteins on the surface of the membrane to a certain extent, thereby effectively improving the service life and the working efficiency of the ultrafiltration membrane.
Detailed Description
The present invention will be described in detail with reference to the following specific examples:
the invention relates to an ultrafiltration purification system for porcine reproductive and respiratory syndrome virus, which comprises a first concentration tank, a second concentration tank and a third concentration tank which are sequentially connected through pipelines, wherein a micro-filter is arranged in front of the first concentration tank, a first ultrafiltration membrane package is arranged between the first concentration tank and the second concentration tank, a second ultrafiltration membrane package is arranged between the second concentration tank and the third concentration tank, a first ultrafiltration membrane and a second ultrafiltration membrane are arranged in the first ultrafiltration membrane package and the second ultrafiltration membrane package Bie An, the pore diameter of the first ultrafiltration membrane is larger than the molecular weight of target virus, the pore diameter of the second ultrafiltration membrane is smaller than the molecular weight of target virus, specifically, the pore diameter of the first ultrafiltration membrane is 35-40nm, the pore diameter of the second ultrafiltration membrane is 15-25nm, and the first ultrafiltration membrane and the second ultrafiltration membrane are both modified polyether sulfone ultrafiltration membranes. The method comprises the following steps:
example 1
Preparation of modified montmorillonite
Stirring 25g of montmorillonite, dispersing in 500ml of 0.9wt% physiological saline, heating to boiling, cooling to room temperature, filtering, washing, dispersing in 600ml of deionized water, adding 10ml of 5wt% tetramethyl ammonium chloride solution dropwise, reacting for 4 hours at 80-90 ℃, filtering, washing, drying, and grinding to obtain modified montmorillonite.
Preparation of composite modified particles
S1: stirring and dispersing 20g of modified montmorillonite in 150mL of acetone solution, adding 5g of acrylic acid monomer, stirring and mixing uniformly by magnetic force, adding 0.5g of hexadecyl trimethyl ammonium bromide and 0.2g of benzoyl peroxide amine, heating to 110 ℃ in an oil bath after stirring and mixing uniformly, preserving heat and reacting for 100min, and then removing acetone in vacuum at room temperature to obtain a material I;
s2: stirring and dispersing 12g of nano cellulose crystal in 100mL of toluene, performing ultrasonic dispersion to obtain a suspension, placing the mixed material I in a ball mill, adding the suspension, performing ball milling for 30min at the speed of 300rpm, taking out, recovering toluene under reduced pressure, drying the obtained solid, stirring in the heating process from far infrared heating to 70 ℃, spraying 20mL of 2wt% chitosan acetic acid solution, continuously stirring for 10-15min after spraying, and drying to obtain the composite modified particles.
The pretreatment of the composite modified particles is as follows: 10g of composite modified particles are stirred and dispersed in 200mL of deionized water, 1g of ammonium cerium nitrate and 20mL of concentrated sulfuric acid are added, the stirring reaction is continued for 1-2h at normal temperature, the filtration is carried out after the reaction is completed, and a filter cake is washed to be neutral and dried.
Respectively weighing 100 parts of polyethersulfone, 3 parts of polyvinylpyrrolidone, 18 parts of composite modified particles and 4 parts of dimethylacetamide according to the proportion, adding the pretreated composite modified particles into dimethylacetamide, performing ultrasonic dispersion for 30min, adding 2 parts of ammonium persulfate and 1 part of sodium dodecyl benzoate after stirring and dissolving, continuously introducing nitrogen for 30min, heating to 90 ℃ under nitrogen atmosphere, performing heat preservation and stirring reaction for 5h, then adding polyvinylpyrrolidone, continuously stirring and reacting for 2h to obtain a casting solution, stopping stirring, performing heat preservation and standing for deaeration for 6h, cooling to room temperature, scraping a film, performing cooling and molding, then soaking in distilled water for 3d, and drying to obtain the modified polyethersulfone ultrafiltration film.
The prepared modified polyethersulfone ultrafiltration membrane is used for purifying porcine reproductive and respiratory syndrome virus in a purification system, wherein the removal rate of invalid proteins is 96 percent, the contact angle of the tested modified polyethersulfone ultrafiltration membrane is 104.9 degrees, and the flux recovery rate after entrapping bovine serum albumin reaches 83.6 percent.
Example two
The modified montmorillonite was prepared as in example one.
Preparation of composite modified particles
S1: stirring and dispersing 20g of modified montmorillonite in 150mL of acetone solution, adding 8g of acrylic acid monomer, stirring and mixing uniformly by magnetic force, adding 0.8g of hexadecyl trimethyl ammonium bromide and 0.2g of benzoyl peroxide amine, heating to 120 ℃ in an oil bath after stirring and mixing uniformly, preserving heat and reacting for 90min, and then removing acetone in vacuum at room temperature to obtain a material I;
s2: stirring and dispersing 11g of nano cellulose crystal in 100mL of toluene, performing ultrasonic dispersion to obtain a suspension, placing the mixed material I in a ball mill, adding the suspension, performing ball milling for 45min at the speed of 300rpm, taking out, recovering toluene under reduced pressure, drying the obtained solid, stirring in the heating process from far infrared heating to 75 ℃, spraying 20mL of 1wt% chitosan acetic acid solution, continuously stirring for 10-15min after spraying, and drying to obtain the composite modified particles.
The pretreatment of the composite modified particles is as follows: 15g of the composite modified particles are stirred and dispersed in 200mL of deionized water, 1.2g of ammonium cerium nitrate and 22mL of concentrated sulfuric acid are added, the stirring reaction is continued for 1h at normal temperature, the filtration is carried out after the reaction is completed, and the filter cake is washed to be neutral and dried.
Respectively weighing 95 parts of polyethersulfone, 5 parts of polyvinylpyrrolidone, 20 parts of composite modified particles and 3 parts of dimethylacetamide according to the proportion, adding the pretreated composite modified particles into dimethylacetamide, performing ultrasonic dispersion for 45min, adding 2 parts of ammonium persulfate and 1 part of sodium dodecyl benzoate after stirring and dissolving, continuously introducing nitrogen for 30min, heating to 80 ℃ under nitrogen atmosphere, performing heat preservation and stirring reaction for 3h, then adding polyvinylpyrrolidone, heating to 85 ℃, continuously stirring and reacting for 3h, obtaining casting solution, stopping stirring, performing heat preservation and standing for deaeration for 6h, cooling to room temperature, scraping a film, performing cooling and molding, then soaking in distilled water for 2d, and drying to obtain the modified polyethersulfone ultrafiltration membrane.
The prepared modified polyethersulfone ultrafiltration membrane is used for purifying porcine reproductive and respiratory syndrome virus in a purification system, wherein the removal rate of invalid proteins is 95%, the contact angle of the tested modified polyethersulfone ultrafiltration membrane is 102.6 degrees, and the flux recovery rate after entrapping bovine serum albumin reaches 82.7%.
Example III
The modified montmorillonite was prepared as in example one.
Preparation of composite modified particles
S1: stirring and dispersing 20g of modified montmorillonite in 150mL of acetone solution, adding 10g of acrylic acid monomer, stirring and mixing uniformly by magnetic force, adding 0.6g of hexadecyl trimethyl ammonium bromide and 0.1g of benzoyl peroxide amine, heating to 100 ℃ in an oil bath after stirring and mixing uniformly, preserving heat and reacting for 120min, and then removing acetone in vacuum at room temperature to obtain a material I;
s2: stirring and dispersing 10g of nano cellulose crystal in 100mL of toluene, performing ultrasonic dispersion to obtain a suspension, placing the mixed material I in a ball mill, adding the suspension, performing ball milling for 40min at the speed of 250rpm, taking out, recovering toluene under reduced pressure, drying the obtained solid, stirring in the heating process from far infrared heating to 70 ℃, spraying 20mL of 1wt% chitosan acetic acid solution, continuously stirring for 10-15min after spraying, and drying to obtain the composite modified particles.
The pretreatment of the composite modified particles is as follows: 13g of composite modified particles are stirred and dispersed in 200mL of deionized water, 1.1g of ammonium cerium nitrate and 25mL of concentrated sulfuric acid are added, stirring reaction is continued for 2h at normal temperature, filtering is carried out after the reaction is completed, and filter cakes are washed to be neutral and dried.
Respectively weighing 95 parts of polyethersulfone, 4 parts of polyvinylpyrrolidone, 15 parts of composite modified particles and 2 parts of dimethylacetamide according to the proportion, adding the pretreated composite modified particles into dimethylacetamide, performing ultrasonic dispersion for 40min, adding 2 parts of ammonium persulfate and 1 part of sodium dodecyl benzoate after stirring and dissolving, continuously introducing nitrogen for 30min, heating to 95 ℃ under nitrogen atmosphere, performing heat preservation and stirring reaction for 4h, then adding polyvinylpyrrolidone, heating to 90 ℃, continuously stirring and reacting for 2h, obtaining casting solution, stopping stirring, performing heat preservation and standing for deaeration for 6h, cooling to room temperature, scraping a film, performing cooling and molding, then soaking in distilled water for 3d, and drying to obtain the modified polyethersulfone ultrafiltration membrane.
The prepared modified polyethersulfone ultrafiltration membrane is used for purifying porcine reproductive and respiratory syndrome virus in a purification system, wherein the removal rate of invalid proteins is 97%, the contact angle of the tested modified polyethersulfone ultrafiltration membrane is 105.8 degrees, and the flux recovery rate after entrapping bovine serum albumin reaches 86.4%.
The above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications and equivalents may be made thereto without departing from the spirit and scope of the technical solution of the present invention, which is intended to be covered by the scope of the claims of the present invention. The technology, shape, and construction parts of the present invention, which are not described in detail, are known in the art.

Claims (5)

1. The ultrafiltration purification system for the porcine reproductive and respiratory syndrome virus is characterized by comprising a first concentration tank, a second concentration tank and a third concentration tank which are sequentially connected through pipelines, wherein a micro filter is arranged in front of the first concentration tank, a first ultrafiltration membrane package is arranged between the first concentration tank and the second concentration tank, a second ultrafiltration membrane package is arranged between the second concentration tank and the third concentration tank, a first ultrafiltration membrane and a second ultrafiltration membrane are arranged in the first ultrafiltration membrane package and the second ultrafiltration membrane package Bie An, the pore diameter of the first ultrafiltration membrane is larger than the molecular weight of a target virus, and the pore diameter of the second ultrafiltration membrane is smaller than the molecular weight of the target virus;
the first ultrafiltration membrane and the second ultrafiltration membrane are both modified polyethersulfone ultrafiltration membranes, and the modified polyethersulfone ultrafiltration membranes comprise the following raw materials in parts by weight: 95-100 parts of polyethersulfone, 3-5 parts of polyvinylpyrrolidone, 15-20 parts of composite modified particles and 2-4 parts of dimethylacetamide;
the composite modified particles are formed by modifying modified montmorillonite through polyacrylic acid intercalation and then compositing the modified montmorillonite with nano cellulose crystals;
the preparation method of the modified polyethersulfone ultrafiltration membrane comprises the following steps: adding pretreated composite modified particles into dimethylacetamide, performing ultrasonic dispersion for 30-45min, adding polyether sulfone, stirring and dissolving, adding ammonium persulfate and sodium dodecyl benzoate, continuously introducing nitrogen for 30min, heating to 80-95 ℃ under nitrogen atmosphere, performing heat preservation and stirring reaction for 3-5h, then adding polyvinylpyrrolidone, continuously stirring and reacting for 2-3h to obtain a casting solution, stopping stirring, performing heat preservation and standing and defoaming for 6h, cooling to room temperature, scraping a film, performing cooling and molding, then soaking in distilled water for 2-3d, and drying to obtain a modified polyether sulfone ultrafiltration membrane;
the pretreatment of the composite modified particles comprises the following steps: stirring and dispersing the composite modified particles in deionized water, adding ammonium ceric nitrate and concentrated sulfuric acid, continuously stirring at normal temperature for reaction for 1-2h, filtering after the reaction is finished, and washing a filter cake to be neutral and drying.
2. The porcine reproductive and respiratory syndrome virus ultrafiltration purification system according to claim 1, wherein the pore size of the first ultrafiltration membrane is 35-40nm and the pore size of the second ultrafiltration membrane is 15-25nm.
3. The porcine reproductive and respiratory syndrome virus ultrafiltration purification system according to claim 1, wherein the preparation method of the composite modified particles is as follows:
s1: stirring and dispersing the modified montmorillonite in an acetone solution, adding an acrylic acid monomer, stirring and mixing uniformly by magnetic force, adding hexadecyl trimethyl ammonium bromide and benzoyl peroxide amine, stirring and mixing uniformly, heating to 100-120 ℃ in an oil bath, reacting for 90-120min at a constant temperature, and then removing acetone in vacuum at room temperature to obtain a material I;
s2: dispersing nanocellulose crystal in toluene by stirring, ultrasonically dispersing to obtain a suspension, placing the mixed material I in a ball mill, adding the suspension, ball milling for 30-45min at the speed of 200-300rpm, taking out, decompressing, recovering toluene, drying to obtain solid, stirring in the heating process of far infrared heating to 70-75 ℃, simultaneously spraying 1-2wt% chitosan acetic acid solution, continuing stirring for 10-15min after spraying, and drying to obtain composite modified particles.
4. A porcine reproductive and respiratory syndrome virus ultrafiltration purification system according to claim 3, wherein the modified montmorillonite is prepared by the following steps: stirring and dispersing montmorillonite in 0.9wt% physiological saline, heating to boiling, cooling to room temperature, filtering and washing, stirring and dispersing in deionized water, dripping tetramethylammonium chloride solution, reacting for 4 hours at 80-90 ℃, and carrying out suction filtration, washing, drying and grinding after the reaction is completed to obtain the modified montmorillonite.
5. The ultrafiltration purification system for porcine reproductive and respiratory syndrome virus according to claim 4, wherein in the preparation of the modified montmorillonite, the mass ratio of montmorillonite to tetramethylammonium chloride solution is 1 (0.2-0.5).
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