CN114561803B - Virus trapping material and production process thereof - Google Patents
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- CN114561803B CN114561803B CN202111456480.9A CN202111456480A CN114561803B CN 114561803 B CN114561803 B CN 114561803B CN 202111456480 A CN202111456480 A CN 202111456480A CN 114561803 B CN114561803 B CN 114561803B
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- D06M13/184—Carboxylic acids; Anhydrides, halides or salts thereof
- D06M13/207—Substituted carboxylic acids, e.g. by hydroxy or keto groups; Anhydrides, halides or salts thereof
-
- D—TEXTILES; PAPER
- D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
- D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
- D06M15/00—Treating fibres, threads, yarns, fabrics, or fibrous goods made from such materials, with macromolecular compounds; Such treatment combined with mechanical treatment
- D06M15/01—Treating fibres, threads, yarns, fabrics, or fibrous goods made from such materials, with macromolecular compounds; Such treatment combined with mechanical treatment with natural macromolecular compounds or derivatives thereof
- D06M15/03—Polysaccharides or derivatives thereof
- D06M15/05—Cellulose or derivatives thereof
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- D—TEXTILES; PAPER
- D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
- D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
- D06M15/00—Treating fibres, threads, yarns, fabrics, or fibrous goods made from such materials, with macromolecular compounds; Such treatment combined with mechanical treatment
- D06M15/01—Treating fibres, threads, yarns, fabrics, or fibrous goods made from such materials, with macromolecular compounds; Such treatment combined with mechanical treatment with natural macromolecular compounds or derivatives thereof
- D06M15/03—Polysaccharides or derivatives thereof
- D06M15/05—Cellulose or derivatives thereof
- D06M15/07—Cellulose esters
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- D—TEXTILES; PAPER
- D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
- D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
- D06M16/00—Biochemical treatment of fibres, threads, yarns, fabrics, or fibrous goods made from such materials, e.g. enzymatic
- D06M16/003—Biochemical treatment of fibres, threads, yarns, fabrics, or fibrous goods made from such materials, e.g. enzymatic with enzymes or microorganisms
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2101/00—Chemical composition of materials used in disinfecting, sterilising or deodorising
- A61L2101/02—Inorganic materials
- A61L2101/26—Inorganic materials containing copper
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2101/00—Chemical composition of materials used in disinfecting, sterilising or deodorising
- A61L2101/32—Organic compounds
- A61L2101/36—Carboxylic acids or derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2101/00—Chemical composition of materials used in disinfecting, sterilising or deodorising
- A61L2101/32—Organic compounds
- A61L2101/46—Macromolecular compounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2101/00—Chemical composition of materials used in disinfecting, sterilising or deodorising
- A61L2101/32—Organic compounds
- A61L2101/46—Macromolecular compounds
- A61L2101/50—Polysaccharides or derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2202/00—Aspects relating to methods or apparatus for disinfecting or sterilising materials or objects
- A61L2202/20—Targets to be treated
- A61L2202/26—Textiles, e.g. towels, beds, cloths
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2209/00—Aspects relating to disinfection, sterilisation or deodorisation of air
- A61L2209/10—Apparatus features
- A61L2209/14—Filtering means
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- D—TEXTILES; PAPER
- D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
- D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
- D06M2101/00—Chemical constitution of the fibres, threads, yarns, fabrics or fibrous goods made from such materials, to be treated
- D06M2101/16—Synthetic fibres, other than mineral fibres
- D06M2101/18—Synthetic fibres consisting of macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds
- D06M2101/20—Polyalkenes, polymers or copolymers of compounds with alkenyl groups bonded to aromatic groups
Abstract
The application discloses a virus trapping material and a production process thereof, wherein the virus trapping material comprises a supporting material layer, a coating layer and a grafting modification layer, and the supporting material layer is a medical non-woven fabric air filtering material; coating a coating layer with the thickness not more than 50nm on the outer layer of the supporting material; the grafting modification layer is coated on one side of the coating layer far away from the supporting material layer, and the thickness of the grafting modification layer is not more than 10nm; the coating comprises 1000 parts of gelatin and 1000 parts of Cu/Cu by weight 2+ 0.5-3 parts, 40-200 parts of cellulose and 50-800 parts of glycyrrhizic acid; 1 part of fibronectin and 0.05-3 parts of RNaseA enzyme by weight of the grafting modification layer; the grafting modification layer is used for specific affinity adsorption and capturing virus particles passing through the vicinity of the micropores and then inactivating viruses in cooperation with the coating layer. The material can effectively trap virus and combine copperThe ions are capable of inactivating the virus simultaneously.
Description
Technical Field
The invention relates to a virus trapping material and application thereof, in particular to a material for specifically trapping and removing surface modification and coating of viruses and application of the virus trapping material in virus detection.
Background
The main principle of the existing air filtering materials, such as various non-woven fabrics materials and antibacterial and antiviral materials, such as copper oxide non-woven fabrics, is that the suspended particles in aerosol are trapped by impact sedimentation and other physical or dynamic modes to play a role in deep filtration, so that the trapping effect of suspended particles with different particle sizes is not linear, for example, when the test is carried out by using a non-woven fabric filter disc of an N95 surgical mask, the trapping rate of micron-sized bacteria particles is larger than that of spherical particles with particle sizes of about 60-150nm, such as influenza viruses, and the trapping rate ratio of the particles is larger than 1. The use of such air filtration materials, even in national standards or ASTM standard-accepted membranes, is potentially risky for blocking infectious viral particles, such as Cov229E, cov19, etc. of the coronavirus type, since this simple physical entrapment does not have selective entrapment and inactivation of biological properties for certain dangerous pathogens.
Therefore, the capability of selectively adsorbing and capturing specific pathogens is increased by further modifying the materials, the conventional monoclonal antibody surface grafting process is expensive in raw materials, and a plurality of factors are considered, so that a fixing process flow is carefully designed for retaining the activity of the monoclonal antibody; another option is to use a low-cost gelatin coating or several lysine polymer coatings, some toxic bifunctional crosslinking reagents are needed in the process operation, and the thickness of the coating is not easy to control in the actual operation, and is a major problem, so that defects of reduced aperture ratio, ventilation parameters and the like of a filter membrane often occur when a qualified specific surface area of the coating is pursued, so that more precise membrane manufacturing equipment needs to be specially developed and skills of operators are trained, and the production cost is too high.
In addition, the use of directly coated copper oxide in the filter material, while improving pathogen killing efficiency, brings about Cu 2+ Dissociation release of ions, e.g. drinking water free Cu 2+ Ions exceeding 2ppm can present a safety hazard. If the nano copper is not closely attached to the mask fiber during the production of the mask, the mask can be directly inhaled when worn. Especially when copper oxide particles are small to nanometer units, the penetration power of the copper oxide particles is stronger, and the nanometer particles can generate genotoxicity after being inhaled, thereby bringing crisis to health; the suspension of copper oxide nanoparticles in air will have the opportunity to be inhaled by the human body, if a large number of foreign objects have destroyed the lung tissue, resulting in pulmonary fibrosis.
In addition, cu in the copper oxide coating is directly coated 2+ The principle of pathogen killing is that copper oxide, especially Cu 2+ Cu which is easily adsorbed by pathogen structural protein through chelating coordination 2+ Concentrations above 1-10ppm can cause membrane structural disruption of pathogens in the course of minutes to tens of minutes. In order to achieve this function, the filter material must also satisfy two conditions: first, the pathogen is in close contact with the filter material to ensure substantial Cu 2+ Ion mass transfer, while also requiring sufficient Cu 2+ Mass transfer time. None of the copper oxide air filtration materials and processes reported so far achieve these characteristics.
Disclosure of Invention
According to the embodiment of the application, the problem that the non-woven fabric cannot intercept and inactivate biological characteristics of viruses in the prior art is solved by providing the virus catching material and the production process, and the effect that the non-woven fabric air filter material can intercept and inactivate the viruses biologically is achieved.
The embodiment of the application provides a virus trapping material, which comprises a supporting material layer, a coating layer and a grafting modification layer, wherein the supporting material layer is a medical non-woven fabric air filtering material;
coating a coating layer with the thickness not more than 50nm on the outer layer of the supporting material;
the grafting modification layer is coated on one side of the coating layer far away from the supporting material layer, and the thickness of the grafting modification layer is not more than 10nm;
the coating comprises 1000 parts of gelatin, 0.5-3 parts of Cu/Cu < 2+ >, 40-200 parts of cellulose and 50-800 parts of glycyrrhizic acid in parts by weight;
1 part of fibronectin and 0.05-3 parts of RNaseA enzyme by weight of the grafting modification layer;
the grafting modification layer is used for specific affinity adsorption and capturing virus particles passing through the vicinity of the micropores and then inactivating viruses in cooperation with the coating layer.
Furthermore, the coating layer adopts modified gelatin protein and cellulose/nitrocellulose which are subjected to covalent aging treatment and then play a virus inactivation role with chelated Cu/Cu < 2+ > and glycyrrhizic acid;
the fibronectin and RNaseA of the grafting layer are connected and assembled with the gelatin protein and cellulose/nitrocellulose of the coating layer in a non-covalent mode through a self-assembly mode, and the trapped virus particles are inactivated by the cooperation of the coating layer.
Further, the concentration of the prepared coating layer spraying solution is 20-200mg/L of gelatin, 0.05-5mg/L of copper sulfate/copper oxide, 5-50 mg/L of cellulose/nitrocellulose and 5-80 mg/L of glycyrrhizic acid;
the concentration of the grafting layer spraying solution ranges from 1.0ug to 2.0mg/L of fibronectin and from 1.0ug to 5.0mg/L of RNase A.
Further, copper or copper ions are added in an amount of not more than 1 mmole per kg of nonwoven fabric.
Furthermore, the simple substance form of Cu/Cu < 2+ > in the constructed coating layer is embedded in the gelatin/glycyrrhizic acid/cellulose coating layer, and the ionic form of Cu/Cu < 2+ > is combined with amino acid residues in a non-free mode to serve as a catalyst for catalyzing local oxidation reaction; the N end of the fibronectin of the constructed graft layer is fixed on the gelatin protein of the bottom layer in a self-assembly mode, and the RGD sequence of the fibronectin is combined with the S protein of the virus particle in an affinity adsorption mode.
A production process comprising the steps of:
step one, dip-coating and spraying the surface of the non-woven fabric with Cu < 2+ > (0.05-5 ppm copper sulfate/copper oxide) gelatin coating liquid, centrifuging the compressed air or hydrogen to remove redundant dip-coating liquid, aging, and carrying out reduction treatment;
and step two, modifying with a grafting solution containing (2 ug/L-2 mg/L) fibronectin, RNase A, glycyrrhizic acid and nitrocellulose, and spraying the prepared solution onto the surface of the coating layer formed in the step one.
The components form a composite structure by a self-assembly mode, and the assembly sequence of the gel sol is as follows: the gelatin molecules at the bottom layer are solidified and embedded with copper sulfate on the surface of a matrix material in a covalent way, nitrocellulose/cellulose and gelatin molecules are assembled into a mesh structure in an electrostatic adsorption way, the N end of fibronectin and gelatin molecules are assembled into a complex in an affinity adsorption way, the C end of fibronectin is in a free state, and the grafting process is completed in a way of combining an intermediate structure of the N end and the C end with RNase A. .
One or more technical solutions provided in the embodiments of the present application at least have the following technical effects or advantages:
by adopting the fibronectin composite virus inactivating component, the problem that the non-woven fabric in the prior art can not effectively selectively intercept viruses is effectively solved, and the effect that the non-woven fabric can intercept and inactivate the viruses is further realized.
The modification layer prepared by the process is firmly combined, the chelated Cu < 2+ > ions are mainly embedded and stored in the gelatin in the form of simple substance Cu, and a large amount of active Cu < 2+ > is generated only after pathogens are captured and obtained, so that potential toxicity does not exist; owing to the safety concentration, cu in the whole process of Cu < 2+ > disinfection 2+ Are in a bound state to chelate with the amino acid residue structure on the coating, so that no free ions are released at all.
After the modification layer prepared by the process captures pathogen viruses in the gas phase, the modification layer is the subsequent Cu 2+ The mass transfer and inactivation process provides enough reaction time, viruses are mutually connected into electronic conductors after being fixed by Fn, and the pathogen anode is automatically triggered by the overpotential of virus bacteria to perform the inactivation process; if the pathogen is not presentHas obvious anode reaction and Cu on the surface of the material 2+ The concentration remains below the safe concentration, so the solution is toxic free Cu 2+ The liquid or gaseous state is not dissociated at all and the liquid or gaseous state is not lost to the environment or human body.
Drawings
FIG. 1 shows a constant temperature micro pressure difference test fixture:
FIG. 2 is a graph showing the pressure drop change test results of a coating solution treatment membrane;
FIG. 3 is a qualitative comparison of the transmittance Rp (%) of the film before and after the treatment with the coating liquid;
FIG. 4 a cyclic voltammogram of a cov229e capture patch;
FIG. 5 shows the results of the membrane S protein adsorption capacity test after modification of the grafting liquid 3.
In the figure, 1. Stainless steel disc filter inlet; 2. testing the membrane; 3. a pressure-bearing micro-pore plate; 4. stainless steel disc filter outlet; 5. and a constant-temperature water bath jacket.
Detailed Description
In order that the invention may be readily understood, a more complete description of the invention will be rendered by reference to the appended drawings; the preferred embodiments of the present invention are illustrated in the drawings, however, the present invention may be embodied in many different forms and is not limited to the embodiments described herein; rather, these embodiments are provided so that this disclosure will be thorough and complete.
It should be noted that the terms "vertical", "horizontal", "upper", "lower", "left", "right", and the like are used herein for illustrative purposes only and do not represent the only embodiment.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs; the terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention; the term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
Fibronectin has a relative molecular weight of about 450KD and is formed of two subunits of about 230KD linked by interchain disulfide bonds. Fibronectin is closely related to cell adhesion, diffusion, proliferation, migration and embryonic development in many metabolic pathways. It is also involved in a variety of pathological processes of tissues such as wound healing, inflammation, fibrosis and modulation of immune cells such as capture and conditioning of coronaviruses, HIV, rabies viruses and a variety of bacteria.
One of the main functions of fibronectin is to mediate cell adhesion. Fibronectin helps regulate cell shape and cytoskeletal organization through cellular signal transduction pathways, and promotes cell diffusion. Numerous studies and clinical applications have demonstrated that FN plays a very important role in mediating cellular and humoral immunity. Depending on the structure of its HIII region, FN molecules can mediate complex coordination of CD4 positive T cells with gp41/gp120 or similar sites in T cell immunocompetence.
A variety of viruses, such as HIV, severe acute respiratory syndrome coronavirus (SARS-CoV) and middle east respiratory syndrome coronavirus (MERS-CoV), all enter the cell through the common receptor CD4 molecule and gp120/gp41 positive membrane protein receptor, while CD4 is widely present on the membrane surface of T cells, and these types of viral particles can enter the host cell in a receptor-like manner. These pathway patterns are generally associated with tight interactions with endothelial cells that have the effect of supporting T cell proliferation and increasing Treg inhibition. This ability of the endothelium to enhance Treg function can provide a target for immune molecules to enhance Treg activity during inflammatory disease. Much evidence suggests that the use of FN or RGD/RGDS peptide-like fragments can interfere with blocking viral infection by a Spacer-Arm ligand-like manner. This results from the specific affinity of FN to various protein molecules of the viral envelope or outer membrane, in other words, one of the inferences that has been made is that it is possible to capture this class of viruses using FN as a material surface structure.
What is known in the Chinese name 2019-nCoV and Covid19 or SARS-CoV is the new coronavirus, 2019-nCoV having an envelope containing the S protein of gp 41-like motif. The particles are round or oval, usually polygonal, with a diameter of 60nm to 140nm. It has more than 85% homology with SARS-CoV. After 96 hours of in vitro isolation and culture 2019-nCoV can be found in human airway epithelial cells, which includes a process in which FN is intimately involved. And there is evidence that infection with 2019-nCoV is also a CD 4-mediated pathway.
The above background materials indicate that various viruses, including novel coronaviruses, whose S protein first performs the function of site-directed binding and that of opening the gate of a host cell is based on its own RGD amino acid motif binding domain, whereas RGD structure is present in large numbers in the Fibronectin (FN) molecular structure; the fibronectin or the enzyme digested fragment thereof from natural sources has a potential function, is possible to repair tissue injury, neutralize and fix, condition viruses and bacteria, has the characteristics of no toxicity and no residue of natural products, and has the characteristics of widely accepted environment, biological safety, high stability and economy.
There are reports and patent applications on treatment and virus prevention based on this principle, but there is no concern about making an air filter material for virus trapping and killing by utilizing this characteristic.
Therefore, the core of the invention is to prepare the surface modification material capable of capturing the viruses with high efficiency and specificity by utilizing the natural complementary characteristics. The invention is characterized in that a layer of nano-scale coating is prepared on the surface of a textile, such as a copper oxide non-woven fabric with disinfection and sterilization functions, by using natural and economic fibrinectin molecules and natural product compositions such as cheap and nontoxic glycyrrhizic acid, RNaseA, cellulose or nitrocellulose derivatives, so as to capture, filter, condition specific viruses and prolong the effective time of virus inactivation; the designed solution composition has a hierarchical structure, and can form a supermolecule composite structure with certain strength on the surface of a material, and after the synergistic effect, the active ingredients can exert the efficacy of effectively inactivating viruses in a low-dose state; in addition, the embedding layer can greatly reduce the release of low-toxic components on the surface of the original substrate, for example, copper oxide particles, nano silver and nano gold fiber particles are prevented from peeling off and losing from the surfaces of non-woven fabric materials and electrode materials.
Example 1
The virus trapping material comprises a supporting material layer, a coating layer and a grafting modification layer, wherein the supporting material layer is a medical non-woven fabric air filtering material;
coating a coating layer with the thickness not more than 50nm on the outer layer of the supporting material;
the grafting modification layer is coated on one side of the coating layer far away from the supporting material layer, and the thickness of the grafting modification layer is not more than 10nm;
the coating comprises 1000 parts of gelatin, 0.5-3 parts of Cu/Cu < 2+ >, 40-200 parts of cellulose and 50-800 parts of glycyrrhizic acid in parts by weight;
1 part of fibronectin and 0.05-1 part of RNaseA enzyme by weight part of the grafting modification layer;
the grafting modification layer is used for specific affinity adsorption and capturing virus particles passing through the vicinity of the micropores and then inactivating viruses in cooperation with the coating layer.
And (3) film making: commercially available N95 polypropylene melt blown nonwoven (specific surface area>12000cm 2 Immersing and cleaning for 2 hours by pretreatment liquid containing 0.1% TritonX100 and pH3.0, installing and fixing a single-layer non-woven fabric membrane (diameter of 90 mm) by a device shown in figure 1 or a clamp with larger size based on similar principle, repeatedly filtering by a peristaltic pump for 15 minutes by feeding the basic coating proportioning liquid into the clamp, and repeatedly purging by connecting compressed air for 3 minutes; taking out the membrane, putting the membrane into a constant temperature and humidity aging box with the humidity of 60% and the temperature of 50 ℃ for aging for 24 hours, then introducing 3% hydrogen, adjusting the humidity to be lower than 40%, cooling to room temperature for processing for 6 hours, then drying the membrane by compressed air flow to serve as a basic coating membrane for standby, and measuring the average thickness of the coating by a microbalance weighing method to obtain the average thickness of 31.3nm.
In actual preparation, auxiliary components can be added in the proportion:
coating ratio: 100mg of gelatin, 50mg of glycyrrhizic acid, 20mg of nitrocellulose, 0.28g of disodium hydrogen phosphate, 1.5g of sodium dihydrogen phosphate, 5.6g of sodium chloride, 0.15mg of copper sulfate, 1mg of malondialdehyde and 1000ml of water for injection at room temperature.
Grafting liquid ratio 1: FN 2mg,RNase A0.1mg, disodium hydrogen phosphate 0.28g, sodium dihydrogen phosphate 1.5g, sodium chloride 5.6g, and water for injection 1000ml.
And (3) film making: a single-layer basic coating film is installed and fixed by using a device shown in figure 1 or a clamp with a larger size based on a similar principle, a peristaltic pump is used for sending the grafting solution ratio 1 into the clamp for repeated filtration for 3 minutes, compressed air is used for repeated blowing and drying for 10 minutes, and then the grafting solution ratio 1 is used for modifying the film for later use, and the average thickness of the coating is increased by 1.2nm measured by a microbalance weighing method.
And the ratio of the grafting liquid is 2: FN 20mg, RNase A10 mg, nano gold particles with the particle size of 5nm 100mg, disodium hydrogen phosphate 0.28g, sodium dihydrogen phosphate 1.5g, sodium chloride 5.6g, potassium chloride 1.4g and water for injection 1000ml.
And (3) film making: a single-layer basic coating film is installed and fixed by using a device shown in figure 1 or a clamp with a larger size based on a similar principle, a peristaltic pump is used for sending the grafting solution proportion 2 into the clamp for repeated filtration for 3 minutes, compressed air is used for repeated blowing and drying for 10 minutes, and then the grafting solution proportion 2 is used as a modified film for standby, and a coating thickness increase value of 3.7nm is measured by a microbalance weighing method.
And the grafting liquid ratio 3: fn 2ug, RNase A5 ug, disodium hydrogen phosphate 0.28g, and sodium dihydrogen phosphate 1.5g were dissolved in 1000ml of water for injection.
And (3) film making: a single-layer basic coating film is installed and fixed by using a device shown in figure 1 or a clamp with a larger size based on a similar principle, a peristaltic pump is used for sending the grafting solution proportion 3 into the clamp for repeated filtration for 3 minutes, compressed air is used for repeated blowing and drying for 10 minutes, the grafting solution proportion 3 is used for modifying the film for later use, and the average thickness of the coating is increased by 0.7nm measured by a microbalance weighing method.
Example two
As shown in fig. 1, a commercially available medical non-woven fabric sample (G1) is used as a control, an initial membrane treated by a coating layer in embodiment one is used for starting, then a grafting solution coating solution (G2) in proportion 1 is used for preparing various sample pieces 10 after coating treatment by a grafting solution coating solution (G3) in proportion 3 (G4), 1 piece, 2 piece and 3 piece of samples (2 in fig. 1) are respectively taken and clamped in a stainless steel disc filter in fig. 1, and a filter inlet (1 in fig. 1) is connected with a constant pressure air source by a PEEK pipe and connected to one port of a micro pressure differential meter (measuring range 0-60 pa); the outlet end of the stainless steel disc filter (4 of FIG. 1) was connected to the other end of the micropressure meter (measuring range 0-60 pa) via a PEEK tube, and the entire stainless steel disc filter was immersed in a constant temperature water bath (constant temperature 30.+ -. 1 ℃ C.) shown in 5 of FIG. 1. During specific test, the air outlet valve is slowly opened, the pressure difference between the air source end and the atmosphere is regulated to be about 50 Pa+/-5 Pa, then the air outlet valve is closed, the air inlet valve is slowly opened to ventilate the filter, and the result is read after the pressure difference meter is stable.
As shown in fig. 2, the single-layer or double-layer aeration static pressure difference of the non-woven fabric material treated by various coating liquid is less than 5Pa, and the pressure difference reaches 8-15 Pa under the condition of filling three layers; as shown in FIG. 2, the single layer of the non-woven fabric material treated by various coating liquids has a double-layer aeration static pressure difference of less than 5Pa, the pressure difference reaches 8-15 Pa when three layers are filled, the non-coating pressure difference of the three layers is 40.5+/-1.2 Pa, the coating 1 is 40.5+/-0.8 Pa, the coating 2 is 40.7+/-2.0 Pa, and the coating 3 is 37.6+/-1.8 Pa.
Example III
The following procedure is operated in the super clean bench. The non-woven fabric membrane manufactured by the membrane with the minimum grafting layer thickness, namely the grafting proportioning liquid 3, is put into the pressure difference detection clamp of figure 1, and the air source inlet is connected with the aerosol generator, so that the sample liquid for preparing the aerosol is the staphylococcus aureus bacterial culture for detection. The outlet of the clamp is connected with a 60mm agar culture plate through a triangular funnel. The concentration of the staphylococcus aureus bacterial culture mother liquor is 6x10 6 CFU/ml, 50-fold dilution with 1xPBS for detection, and adjusting the speed of the atomizing generator fan, keeping 25ml of sample in each sample cup depleted in about 60 minutes. And changing a new sample and an agar detection plate after each atomization spraying.
The above procedure was repeated repeatedly to test 10 outsourced nonwoven fabric membranes and 10 membranes made with the coating solution of ratio 4, respectively, and finally the results of bacterial culture counts for checking membrane leakage rate were shown in table 1:
TABLE 1
The transmittance of untreated membrane to staphylococcus aureus is 56.2% ± 15.4%; compared with the staphylococcus aureus of the membrane treated by the coating liquid of the grafting matching liquid 3, the transmittance of the staphylococcus aureus is 37.9 percent plus or minus 8.9 percent.
Example IV
As in the previous embodiment, the following procedure is operated in the super clean bench. The non-woven fabric membrane manufactured by the grafting ratio 3 is put into the pressure difference detection clamp of figure 1, the air source is connected with the aerosol generator, and the sample liquid for preparing the aerosol is the common cold virus Cov229E cell lysate for detection. The fixture outlet was connected to a 60mm culture dry plate via a triangular funnel and sterilized with 0.1% gelatin plating prior to use. The concentration of the mother liquor of the Cov229E lysate is 4.5x10 5 CFU/ml, 50-fold dilution with 1xPBS for detection, and adjusting the speed of the atomizing generator fan, keeping 25ml of sample in each sample cup depleted in about 60 minutes. After each atomization spraying, a new sample piece and an agar plate are replaced. The specific virus infection detection method comprises accessing 1×10 to each plate 5 MDCK cells are cultured for 48 hours by adding DMEM culture medium containing 1% calf serum after 2 hours, then the cells are washed by 1xPB and treated by membrane rupture liquid containing 0.15% Triton, and TCID50 values of the virus particle content related to each membrane are read by the conversion of a Karber method by using S protein primary antibody and HRP enzyme-labeled secondary antibody chromogenic read plates respectively.
The results of the TCID50 of virus particles leaked from 3 outsourced nonwoven fabric films and 6 films prepared by the coating liquid of the mixture ratio 3 are shown in Table 2:
TABLE 2
The TCID50 results of the above table are converted to a transmittance Rp of the virions through each patch, with Rp of 67.4% ± 19.9% for 3 commercially available patches; the effect of the Rp of the film treated with the coating solution of formulation 3 on the properties of the coating-modified film material is specifically illustrated by FIG. 3, where Rp is 34.7% + -12.41%.
In general, when aerosol particles pass through an air filter material, a part of the particles are blocked by various mechanisms, and a part of the particles can permeate, and the ratio Rp of the permeation portions can be expressed as:。
the residual part (1-Rp) is trapped in the depth filter material, so that the part of particles are completely recovered for inspection, and the method for evaluating the transmittance Rp is relatively simple and reliable, and is further simplified into the method for only inspecting the thickness t of the material in the practical process development h And the reciprocal of the particle size df of the aerosol target particles for detection, the ASTM standard method is to evaluate the sterilization efficiency of the filter material by using Staphylococcus aureus. However, when the filtration efficiency of target particles with different properties needs to be evaluated transversely, the coefficient η needs to be redefined as a vector, which needs to be greatly analyzed and calculated, and after η is added, it can be qualitatively considered that the filtration efficiency Rp of the filter material and the aerosol particle dimension df are not in a linear relationship, for example, a fact that a conventional medical-grade nonwoven fabric of surgical mask material is generally based on the premise that no assumption of abnormality with a linear formula appears when aerosol particles with a size of less than 50nm to 150nm are filtered, so that Φx174 phage is often selected to replace coronavirus for evaluation in practical operation for safety. But when aerosol particles smaller than 150nm are filtered out, the measured Rp value is actually larger than the linear theoretical Rp value when aerosol particles smaller than 150nm and larger than 50nm are filtered out.
The reason for this is repeatedly emphasized that the biological properties of particles suspended in aerosols with particle sizes of around 50-150 nm are often a class of more environmentally safe interest, since such particles are in most cases infectious viral particles including various coronaviruses, as well as some airborne mycoplasma and the like. If the produced aerosol filter material has particle trapping and eliminating capacity lower than that of the product marked in the 50-150 nanometer section, easily neglected loopholes are formed, and the loopholes which are counterintuitive are difficult to pay attention, so that the losses which are difficult to compensate are caused; in the opposite direction, if the designed filter material has selective trapping and elimination effects on the particle size in the 50-150 nanometer range, the phenomenon of inversion of a linear formula should occur when evaluating the Rp ratio of the filter material, namely, if such an instance can be found in the evaluating process, the existence of the selective trapping capability can be directly described, and the vector eta and other material characteristic related coefficients do not need to be described through a great number of complicated parameter measurement and various numerical simulation processes. In short, the inference is true if the above positive result is measured by a simple aerosol release interception test.
FIG. 3 is a graph 1 showing the results of aerosol transmittance test of commercially available medical nonwoven fabrics with Staphylococcus aureus having a particle size of about 300-400nm, where Rp is 56.2% + -15.4%; FIG. 3 is a graph showing the results of aerosol transmittance testing of commercially available medical nonwoven fabrics with Cov229E having a particle size of about 60-120nm, where Rp is 67.4% + -19.9%; FIG. 3 shows the results of aerosol transmittance test of the coating liquid nonwoven fabric of the mixture ratio 4 by using staphylococcus aureus, wherein Rp is 37.9% +/-8.9%; as a result of aerosol transmittance test of the coating liquid nonwoven fabric of the formulation 4 with Cov229E, which is shown in fig. 3, 4, the Rp is 34.7% ± 12.4%, and the ratio Rp (SA)/Rp (Cov) > =1.
Example five
Intercepting the CuO non-woven fabric membrane wafer (with the diameter of 3.5 mm) treated by the coating liquid of the grafting ratio 3 by using a puncher, attaching the wafer to a WE of a BST154 bare gold electrode, using Ag/Cl2 as a counter electrode and a reference electrode, connecting a Stm32f103 singlechip as a serial port communication unit by using an EmStat-Pico module of a potentiostat, and using a Script CV instruction and a DPV instruction for cyclic voltammetry scanning and peak detection. The scanning potential ranges from 1.0V- (-0.35V), the scanning speed is 50mV/S, and the electrolyte solution is 0.1M Na 2 SO 4 0.1xPBS (pH 7.2). Pre-scanning electrode cleaning under nitrogen gas-introducing conditionAfter the subsequent voltammetric measurement, curve 1 of fig. 4 is the result of scanning with a film sample of Cov229E captured, curve 2 of fig. 4 is the result of scanning with a grafted nonwoven film sample without virus aerosol filtration, curve 1, the average redox potential after 5 consecutive scans of curve 2 is approximately between 0.1 and 0.2V, the redox curve, which is a major form approaching a single electron transfer reaction, appears as positive oxidation reaction in curve 2, gradually falls back after 5-10 consecutive scans, decreasing in the direction approaching curve 2. Curves 1 and 2 and the various deformation processes are all approximate to single electron transfer reactions, and are characterized by reactions involving Cu < 2+ > or Cu < + > in a combined state, which indicates that the Cu < + > ions of the solid-phase catalyst are not dissociated and released into the electrolyte liquid phase.
Example six
Preparing a CuO non-woven fabric membrane treated by a coating solution with a grafting ratio of 3 by using a membrane puncher, loading the membrane into a repeatedly usable phi 35mm filter head, connecting a microinjection pump liquid inlet after confirming no leakage, prefiltering a test solution containing 0.5ng/ml S protein, 0.45% NaCl/0.05 x PBS and pH6.5 by using a cellulose acetate filter head with a pore diameter of 1um, slowly pumping 30ml of the test solution into the filter head at a speed of 1 ml/min, stopping intermittently for 1 min every 1 min, and controlling the time for S protein adsorption every time of filtration; the membrane was then eluted 3 times with 1ml of each of the A, B, C3 eluents, and the pooled eluents were collected by centrifugation and tested for S protein content. Wherein the 3 eluents comprise the following components: a, 0.45% NaCl/0.01 x PBS, pH6.5, B, 0.9% NaCl/0.01 x PBS, pH6.5, C, 0.5M NaCl/0.01 x PBS, pH 6.5.
100ul of S protein standard and washing liquid sample are moved into a test plate hole embedded with the secondary antibody, 100ul of elution buffer A is added for 3 times of washing, and the primary antibody is identified by biotinylation. Unbound antibody was washed away, HRP conjugated, TMB developed and measured at 450 nm. S protein adsorption capacity result conversion method: specific adsorption capacity Ad (0.1 ng/cm 2) = (150-100 x eluent S protein concentration (ng/ml))/9.0.
Under the conditions of eluent a: the S protein adsorption capacity Ad (n=8) of the CuO non-woven fabric membrane is 3.5+/-2.0 (0.1 ng/cm < 2 >), and the S protein adsorption capacity Ad (n=8) of the CuO non-woven fabric membrane treated by the coating solution of the mixture ratio 3 is 13.8+/-3.9 (0.1 ng/cm < 2 >;
under the conditions of eluent B: the S protein adsorption capacity Ad (n=8) of the CuO non-woven fabric membrane is 2.1+/-3.3 (0.1 ng/cm < 2 >), and the S protein adsorption capacity Ad (n=8) of the CuO non-woven fabric membrane treated by the coating solution of the mixture ratio 3 is 13.2+/-4.7 (0.1 ng/cm < 2 >;
under the conditions of eluent C: the S protein adsorption capacity Ad (n=8) of the CuO nonwoven fabric membrane is 0.5+ -1.8 (0.1 ng/cm 2), and the S protein adsorption capacity Ad (n=8) of the CuO nonwoven fabric membrane treated with the coating liquid of the mixture ratio 3 is 13.1+ -3.9 (0.1 ng/cm 2).
Claims (7)
1. The virus trapping material is characterized by comprising a supporting material layer, a coating layer and a grafting modification layer, wherein the supporting material layer is a medical non-woven fabric air filtering material;
coating a coating layer with the thickness not more than 50nm on the outer layer of the supporting material;
the grafting modification layer is coated on one side of the coating layer far away from the supporting material layer, and the thickness of the grafting modification layer is not more than 10nm;
the coating comprises 1000 parts of gelatin and 1000 parts of Cu/Cu by weight 2+ 0.5-3 parts, 40-200 parts of cellulose and 50-800 parts of glycyrrhizic acid;
1 part of fibronectin and 0.05-3 parts of RNaseA enzyme by weight of the grafting modification layer;
the grafting modification layer is used for specific affinity adsorption and capturing virus particles passing through the vicinity of the micropores and then inactivating viruses in cooperation with the coating layer.
2. The virus trapping material according to claim 1, wherein the coating layer is formed of denatured gelatin protein, cellulose/nitrocellulose which is covalently aged and chelated with Cu/Cu 2+ Glycyrrhizic acid exerts virus inactivating function;
the fibronectin and RNaseA of the grafting layer are connected and assembled with the gelatin protein and cellulose/nitrocellulose of the coating layer in a non-covalent mode through a self-assembly mode, and the trapped virus particles are inactivated by the cooperation of the coating layer.
3. The virus trapping material according to claim 2, wherein the concentration of the coating layer spraying solution is 20-200mg/L of gelatin, 0.05-5mg/L of copper sulfate/copper oxide, 5-50 mg/L of cellulose/nitrocellulose, and 5-80 mg/L of glycyrrhizic acid;
the concentration of the grafting layer spraying solution ranges from 1.0ug to 2.0mg/L of fibronectin and from 1.0ug to 5.0mg/L of RNase A.
4. A virus-trapping material according to claim 3, wherein copper or copper ions are added in an amount of not more than 1 mmole per kg of nonwoven fabric.
5. The virus-trapping and inactivating material according to any one of claims 1,2, and 4, wherein Cu/Cu in the constructed coating layer 2+ Is embedded in gelatin/glycyrrhizic acid/cellulose coating layer, cu/Cu 2+ All of the ionic forms of (a) are bound with amino acid residues in a non-free manner as catalysts responsible for catalyzing local oxidation reactions; the N end of the fibronectin of the constructed graft layer is fixed on the gelatin protein of the bottom layer in a self-assembly mode, and the RGD sequence of the fibronectin is combined with the S protein of the virus particle in an affinity adsorption mode.
6. A process for producing the virus-trapping material of claim 1, comprising the steps of:
step one, using Cu-containing alloy 2 + Dip-coating and spray-melting the surface of the non-woven fabric by using 0.05-5ppm of copper sulfate/copper oxide gelatin, glycyrrhizic acid and nitrocellulose coating liquid, centrifuging compressed air or hydrogen to remove redundant dip-coating liquid, aging, and carrying out reduction treatment;
and II, modifying with fibronectin, RNase A and grafting liquid containing 2ug/L-2mg/L, and spraying the prepared solution onto the surface of the coating layer formed in the step I.
7. A production process according to claim 6, wherein the components form a composite structure by self-assembly, and the assembly sequence of the gel sol is as follows: the gelatin molecules at the bottom layer are solidified and embedded with copper sulfate on the surface of a matrix material in a covalent way, nitrocellulose/cellulose and gelatin molecules are assembled into a mesh structure in an electrostatic adsorption way, the N end of fibronectin and gelatin molecules are assembled into a complex in an affinity adsorption way, the C end of fibronectin is in a free state, and the grafting process is completed in a way of combining an intermediate structure of the N end and the C end with RNase A.
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| US5798116A (en) * | 1993-07-30 | 1998-08-25 | University College London | Stabilized materials comprised of copper ion-containing fibronectin mats |
| JP2008188082A (en) * | 2007-02-01 | 2008-08-21 | Nisshinbo Ind Inc | Mask |
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