CN116078360B - Adsorption fiber membrane for removing leucocyte and pathogenic antibody and preparation method thereof - Google Patents
Adsorption fiber membrane for removing leucocyte and pathogenic antibody and preparation method thereof Download PDFInfo
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Abstract
The invention discloses an adsorption fiber membrane for removing leucocytes and pathogenic antibodies and a preparation method thereof, and relates to the technical field of biological medicines. Wherein, the adsorption fiber membrane is prepared by preparing a fiber membrane from a blending solution through an electrostatic spinning technology, and then crosslinking and purifying the fiber membrane; the blend solution comprises a fibrous membrane matrix with good biocompatibility, lipoteichoic acid and polyaromatic amino acid, wherein the mass ratio of the fibrous membrane matrix to the lipoteichoic acid to the polyaromatic amino acid is (1-100): (1-10):1. The adsorption fiber membrane has the advantages of good biocompatibility, good clearance effect on white blood cells and antibodies, and low hemolysis rate, and can be used for purifying blood.
Description
Technical Field
The invention relates to the technical field of biological medicine, in particular to an adsorption fiber membrane for removing leucocyte and pathogenic antibody and a preparation method thereof.
Background
Leukocyte removal therapy (LCAP) is a novel blood purification treatment mode, and by selectively adsorbing leukocytes (including neutrophils, lymphocytes and/or monocytes) in peripheral blood, immune attack of excessive inflammatory cells on a patient body is reduced, and pathogenic proteases, oxygen free radicals, cytokines and the like released by the inflammatory cells are reduced, so that the purposes of protecting organs and relieving diseases are achieved. Currently, LCAP is used in clinic to treat refractory Inflammatory Bowel Disease (IBD), rheumatoid Arthritis (RA), inflammatory response syndrome (SIRS) and other autoimmune diseases.
Wherein, the existing white blood cell adsorbent is used for adsorbing white blood cellsThe adsorption column product comprises Adacolumn Ⓡ Adsorption columns and Cellsorba filters. Adacolumn Ⓡ The adsorbent of the adsorption column is cellulose acetate microsphere, and can adsorb leucocytes; the Cellsorba filter is a filter column formed by winding hydrophilic polymer microfiber non-woven fabrics, and can capture and adsorb white blood cells and partial platelets. However, the existing leukocyte adsorption products mainly aim at leukocyte adsorption, have insufficient effect of eliminating pathogenic antibodies, and have limited application range.
Disclosure of Invention
In order to solve the problems that the existing adsorption column products have good adsorption to white blood cells but poor clearance to pathogenic antibodies, the application provides an adsorption fiber membrane for removing white blood cells and pathogenic antibodies and a preparation method thereof.
In a first aspect, the present application provides an adsorption fiber membrane for removing leukocytes and pathogenic antibodies, which adopts the following technical scheme:
an adsorption fiber membrane for removing leucocyte and pathogenic antibody is prepared by preparing fiber membrane from blending solution by electrostatic spinning technique, and then crosslinking and purifying the fiber membrane; wherein the blend solution comprises a fibrous membrane matrix with good biocompatibility, lipoteichoic acid and polyaromatic amino acid, and the mass ratio of the fibrous membrane matrix, the lipoteichoic acid and the polyaromatic amino acid is (1-100): (1-10):1.
The fibrous membrane matrix can be made of agarose, chitosan, dextran, polyvinyl alcohol and other materials with good biocompatibility, and can be suitable for blood purification. Lipoteichoic acid has the function of adhering white blood cells, has strong antigenicity, can be combined with an antibody, and achieves the aim of removing white blood cells and the antibody simultaneously. While polyaromatic amino acids may bind to antibodies using hydrophobic interactions. Under the synergistic effect of the affinity of lipoteichoic acid and the antibody and the hydrophobic effect of the poly-aromatic amino acid and the antibody, the adsorption fiber membrane can be quickly and stably combined with the antibody, and the adsorption effect of the adsorption fiber membrane on various antibodies is effectively improved.
The adsorbent for purifying the leucocytes and the antibodies in the blood is an adsorption fiber membrane, and compared with the microspherical or non-woven fabric type adsorbent, the adsorbent is more beneficial to the smooth entry of the leucocytes and the plasma into and through the adsorption fiber membrane when the adsorption fiber membrane is used for purifying the blood, and can efficiently and fully remove the leucocytes and the antibodies in the blood.
Further preferably, the mass ratio of the fibrous membrane matrix, lipoteichoic acid and polyaromatic amino acid is (4-20): (1-2):1.
Further preferably, the fibrous membrane substrate is polyvinyl alcohol, the average molecular weight of the polyvinyl alcohol is 15-20 ten thousand, and the alcoholysis degree is 88-98%.
The lipoteichoic acid is obtained by inactivating, crushing, extracting, purifying and freeze-drying any one or more strains of lactobacillus, staphylococcus aureus, streptococcus and bacillus subtilis.
Further preferably, the lipoteichoic acid extraction method comprises the following steps:
1. preparing a bacterial suspension;
2. inactivating the bacterial suspension at high temperature to obtain an inactivated bacterial suspension;
3. performing ultrasonic crushing on the inactivated bacteria suspension, and centrifuging to remove bacterial fragments to obtain an extract;
4. adding an extractant into the extract in the step (3), then centrifugally extracting, and collecting an upper extract, namely a lipoteichoic acid crude product;
5. purifying the crude lipoteichoic acid by an ion exchange column, collecting phosphorus-containing eluent, and freeze-drying to obtain lipoteichoic acid.
Further preferably, the poly-aromatic amino acid is selected from poly-aromatic amino acids with average molecular weight of 1000-20000.
Preferably, the polyaromatic amino acid is selected from the group consisting of phenylalanine and derivatives thereof, polytyrosine and derivatives thereof, and polytryptopine and derivatives thereof.
Preferably, the diameter of the fibers in the fiber membrane is 0.5-50 μm, and the thickness of the fiber membrane is 100-1000 μm.
The fiber membrane is formed by interweaving a plurality of fibers, and the strength of the fiber membrane depends on the diameter of the fibers and the thickness of the fiber membrane. When the diameter of the fiber in the fiber membrane and the thickness of the fiber membrane are within the range, the fiber membrane has good strength performance, and the fiber membrane has good permeability, so that blood cells and blood plasma can enter the fiber membrane more easily, thereby being beneficial to further improving the adsorption effect.
In a second aspect, the preparation method of the adsorption fiber membrane for removing leucocytes and pathogenic antibodies in any one of the first aspects of the application adopts the following technical scheme:
a preparation method of an adsorption fiber membrane for removing leucocytes and pathogenic antibodies comprises the following steps:
s1, preparing a blending solution by adopting polyvinyl alcohol, lipoteichoic acid, poly aromatic amino acid and water;
s2, spinning the blending solution by adopting an electrostatic spinning technology, and then collecting and drying to prepare a fiber membrane;
s3, swelling the fiber membrane, and then adding glutaraldehyde solution into the swelled fiber membrane to perform heating crosslinking under alkaline conditions to obtain a crosslinked fiber membrane;
s4, purifying the crosslinked fiber membrane to obtain the adsorption fiber membrane.
Preferably, in S1, respectively preparing a polyvinyl alcohol solution with the concentration of 15-60g/L, a mixed solution of lipoteichoic acid and polyaromatic amino acid, and then mixing the polyvinyl alcohol, the lipoteichoic acid and the polyaromatic amino acid according to the mass ratio of (1-100): 1-10, adding the mixed solution of lipoteichoic acid and poly aromatic amino acid into the polyvinyl alcohol solution, and stirring and mixing uniformly at 40-45 ℃ to obtain the blending solution.
Preferably, S2 adopts a needleless electrostatic spinning technology to spin, the rotating speed of a rotating cylinder is 3-6r/min, the collecting distance is 15-30cm, and the applied voltage is 20-60KV in the spinning process.
Preferably, the temperature of the drying in S2 is controlled between 55 and 65 ℃.
By adopting the process parameters for spinning, the fiber membrane with the fiber diameter of 0.5-50 mu m and the fiber membrane thickness of 100-1000 mu m can be obtained.
Preferably, the temperature of crosslinking in S3 is controlled to be 45-55 ℃ and the crosslinking time is 2-6h.
In summary, the present application at least includes the following beneficial effects:
(1) The adsorption fiber membrane has good biocompatibility, good effect of removing leucocytes and antibodies, and low hemolysis rate;
(2) Through fibrous diameter and control fibrous thickness in the control fibrous membrane, can improve the intensity performance of absorption fibrous membrane, absorption fibrous membrane has permeability concurrently simultaneously, is favorable to further improving the adsorption effect of this application absorption fibrous membrane.
Description of the embodiments
The present application is further described in detail below by means of preferred embodiments.
Lipoteichoic acid preparation
Suspending lactobacillus in water, placing in a constant temperature water bath, heating at 98deg.C for inactivating for 1 hr, performing ultrasonic crushing treatment for 30min, centrifuging at 5000r/min for 30min to remove bacterial fragments, adding equal volume of saturated n-butanol, stirring for 60min, centrifuging at 8000r/min for 30min, and collecting the upper extract to obtain lipoteichoic acid crude product.
Purifying crude lipoteichoic acid by DEAE-Sephacel ion exchange column chromatography, pre-balancing and loading with 0.1 mol ammonium acetate buffer solution with pH of 4.7 to make it pass through the column slowly, eluting lipoteichoic acid bound on the column with 1mol ammonium acetate buffer solution with pH of 4.7, collecting phosphorus-containing eluate, and lyophilizing to obtain lipoteichoic acid.
The lipoteichoic acid used in the examples below is not limited to lipoteichoic acid extracted by the method described above, but commercially available LTA reagents, which are commercially available, may be used.
Examples
Example 1
An adsorption fiber membrane for removing leucocyte and pathogenic antibody, its preparation method comprises the following steps:
step 1 preparation of a polyvinyl alcohol-lipoteichoic acid-Polytryptophan (PVA-LTA-pTR) ternary blend fiber film
0.5g lipoteichoic acid and 0.5g poly tryptophan (pTR, average molecular weight about 2000) were added to 200 mL water and sonicated to prepare a mixed solution, 200 mL 30g/L polyvinyl alcohol (average molecular weight of polyvinyl alcohol: 20 ten thousand, alcoholysis degree: 98%) was further added thereto, and the mixture was stirred and mixed at 40℃for 30 minutes to obtain PVA: LTA: the mass ratio of pTR is 12:1:1 PVA-LTA-pTR blend solution. Adding PVA-LTA-pTR blend solution into a solution tank, building a needleless electrostatic spinning system, rotating a rotating cylinder in the solution tank at a rotating speed of 3r/min, dipping the PVA-LTA-pTR blend solution in the rotating cylinder, collecting the PVA-LTA-pTR blend solution for 15cm, applying a voltage of 25kV, and controlling the environmental conditions at a humidity of 40% and a temperature of 30 ℃. The collected fibrous membranes were dried in a vacuum oven at 60℃for 24h.
Step 2 crosslinked fibrous membranes
5.0g of dried fibrous membrane is taken out, put in 30 ℃ water for swelling for 2 hours, then taken out and put in 500mL of 0.5% glutaraldehyde solution, 20mL of 1mol/L K is added 2 CO 3 Crosslinking the solution in an oscillating water bath kettle at 50 ℃ for 4 hours to obtain the polyvinyl alcohol-lipoteichoic acid-poly tryptophan (PVA-LTA-pTR) ternary blending adsorption fiber membrane capable of being used for adsorbing leucocytes and pathogenic antibodies. The ternary blending adsorption fiber film of polyvinyl alcohol-lipoteichoic acid-poly tryptophan (PVA-LTA-pTR) is subjected to purification treatment in a Soxhlet extraction mode, namely the adsorption fiber film is placed in a Soxhlet extractor, the adsorption fiber film is extracted for 4 hours by absolute ethyl alcohol and then taken out, purified water is washed until no alcohol residue exists, then the adsorption fiber film is extracted for 6 hours by the purified water, and finally the adsorption fiber film is taken out and placed in a vacuum drying oven for drying 24h at 60 ℃ for later use.
Example 2
An adsorption fiber membrane for removing leucocyte and pathogenic antibody, its preparation method comprises the following steps:
step 1 preparation of a polyvinyl alcohol-lipoteichoic acid-Polytryptophan (PVA-LTA-pTR) ternary blend fiber film
1.0g lipoteichoic acid and 0.5g polytryptophan (pTR, average molecular weight about 2000) are added into 200 mL water to prepare a mixed solution by ultrasonic treatment, 200 mL of 20 g/L polyvinyl alcohol (average molecular weight of polyvinyl alcohol is 20 ten thousand, alcoholysis degree is 98%) solution is added, and stirring and mixing are carried out for 30min at 40 ℃, so that PVA is obtained: LTA: the mass ratio of pTR is 8:2:1 PVA-LTA-pTR blend solution. Adding PVA-LTA-pTR blend solution into a solution tank, building a needleless electrostatic spinning system, rotating a rotating cylinder in the solution tank at a rotating speed of 3r/min, dipping the PVA-LTA-pTR blend solution in the rotating cylinder, collecting the PVA-LTA-pTR blend solution for 15cm, applying a voltage of 30 kV, and controlling the environmental conditions at a humidity of 40% and a temperature of 30 ℃. The collected fiber membranes were dried in a vacuum oven at 60℃for 24h.
Step 2 crosslinked fibrous membranes
5.0. 5.0g of the dried fibrous membrane is swelled in water at 30 ℃ for 2h, then taken out and placed in 500mL of 0.5% glutaraldehyde solution, and 20mL of 1mol/L K is added 2 CO 3 Crosslinking the solution in an oscillating water bath kettle at 50 ℃ for 4 hours to obtain the polyvinyl alcohol-lipoteichoic acid-poly tryptophan (PVA-LTA-pTR) ternary blending adsorption fiber membrane capable of being used for adsorbing leucocytes and pathogenic antibodies. The ternary blending adsorption fiber film of polyvinyl alcohol-lipoteichoic acid-poly tryptophan (PVA-LTA-pTR) is subjected to purification treatment in a Soxhlet extraction mode, namely the adsorption fiber film is placed in a Soxhlet extractor, the adsorption fiber film is extracted for 4 hours by absolute ethyl alcohol and then taken out, purified water is washed until no alcohol residue exists, then the adsorption fiber film is extracted for 6 hours by the purified water, and finally the adsorption fiber film is taken out and placed in a vacuum drying oven for drying 24h at 60 ℃ for later use.
Example 3
An adsorption fiber membrane for removing leucocyte and pathogenic antibody, its preparation method comprises the following steps:
step 1 preparation of a polyvinyl alcohol-lipoteichoic acid-phenylalanine (PVA-LTA-pPH) ternary blend fiber film
Adding 0.5g lipoteichoic acid and 0.5g phenylalanine (pPH, average molecular weight about 1500) into 200 mL water, performing ultrasonic treatment to prepare a mixed solution, adding 200 mL of 15 g/L polyvinyl alcohol (average molecular weight of polyvinyl alcohol is 15 ten thousand, alcoholysis degree is 88%) solution, stirring and mixing for 30min at 40 ℃, and obtaining PVA at the moment: LTA: the mass ratio of pPH is 6:1:1 PVA-LTA-pPH blend solution. Adding PVA-LTA-pPH blend solution into a solution tank, building a needleless electrostatic spinning system, rotating a rotating cylinder in the solution tank at a rotating speed of 3r/min, dipping the PVA-LTA-pPH blend solution in the rotating cylinder, collecting the PVA-LTA-pPH blend solution for 15cm, applying voltage of 20 kV, and controlling environmental conditions at humidity of 40% and temperature of 30 ℃. The collected fiber membranes were dried in a vacuum oven at 60℃for 24h.
Step 2 crosslinked fibrous membranes
5.0. 5.0g of the dried fibrous membrane is swelled in water at 30 ℃ for 2h, then taken out and placed in 500mL of 0.5% glutaraldehyde solution, and 20mL of 1mol/L K is added 2 CO 3 Crosslinking the solution in an oscillating water bath kettle at 50 ℃ for 4 hours to obtain the polyvinyl alcohol-lipoteichoic acid-polyaniline (PVA-LTA-pPH) ternary blending adsorption fiber membrane capable of adsorbing leucocytes and pathogenic antibodies. The ternary blending adsorption fiber film of polyvinyl alcohol-lipoteichoic acid-polyaniline (PVA-LTA-pPH) is subjected to purification treatment in a Soxhlet extraction mode, namely the adsorption fiber film is placed in a Soxhlet extractor, the adsorption fiber film is extracted for 4 hours by absolute ethyl alcohol and then taken out, purified water is washed until no alcohol residue exists, then the adsorption fiber film is extracted for 6 hours by the purified water, and finally the adsorption fiber film is taken out and placed in a vacuum drying oven for drying 24h at 60 ℃ for later use.
Example 4
An adsorption fiber membrane for removing leucocyte and pathogenic antibody, its preparation method comprises the following steps:
step 1 preparation of a polyvinyl alcohol-lipoteichoic acid-poly tyrosine (PVA-LTA-pTyr) ternary blend fiber membrane
Adding 0.5g lipoteichoic acid and 0.5g poly (tyrosine) (pTyr, average molecular weight about 2000) into 200 mL water, performing ultrasonic treatment to prepare a mixed solution, adding 200 mL of 15 g/L polyvinyl alcohol (average molecular weight of polyvinyl alcohol is 15 ten thousand, alcoholysis degree is 88%) solution, stirring and mixing for 30min at 40 ℃, and obtaining PVA at the moment: LTA: the mass ratio of pTyr is 6:1:1 PVA-LTA-pTyr blend solution. Adding PVA-LTA-pTyr blending solution into a solution tank, building a needleless electrostatic spinning system, rotating a rotating cylinder in the solution tank at a rotating speed of 3r/min, dipping the PVA-LTA-pTyr blending solution in the rotating cylinder, collecting the PVA-LTA-pTyr blending solution for 15cm, applying a voltage of 20 kV, and controlling the environmental conditions at a humidity of 40% and a temperature of 30 ℃. The collected fiber membranes were dried in a vacuum oven at 60℃for 24h.
Step 2 crosslinked fibrous membranes
5.0. 5.0g of the dried fibrous membrane is swelled in water at 30 ℃ for 2h, then taken out and placed in 500mL of 0.5% glutaraldehyde solution, and 20mL of 1mol/L K is added 2 CO 3 Crosslinking the solution in an oscillating water bath kettle at 50 ℃ for 4 hours to obtain the polyvinyl alcohol-lipoteichoic acid-poly tyrosine (PVA-LTA-pTyr) ternary blending adsorption fiber membrane capable of being used for adsorbing leucocytes and pathogenic antibodies. The ternary blending adsorption fiber film of polyvinyl alcohol-lipoteichoic acid-poly tyrosine (PVA-LTA-pTyr) is subjected to purification treatment by adopting a Soxhlet extraction mode, namely the adsorption fiber film is placed in a Soxhlet extractor, the adsorption fiber film is extracted for 4 hours by absolute ethyl alcohol and then taken out, purified water is washed until no alcohol residue exists, then the adsorption fiber film is extracted for 6 hours by the purified water, and finally the adsorption fiber film is taken out and placed in a vacuum drying box for drying at 60 ℃ for 24h for later use.
Comparative example 1
An adsorbed fibrous membrane for eliminating leukocytes and pathogenic antibodies, which is different from example 1 in that: the blending solution is not added with poly tryptophan, namely the dosage of lipoteichoic acid is 1.0g, and other parameters are unchanged, thus preparing the polyvinyl alcohol-lipoteichoic acid (PVA-LTA) binary blending adsorption fiber membrane.
Comparative example 2
An adsorbed fibrous membrane for eliminating leukocytes and pathogenic antibodies, which is different from example 1 in that: the blending solution is not added with lipoteichoic acid, namely the dosage of the poly tryptophan is 1.0g, and other parameters are unchanged, so that the polyvinyl alcohol-poly tryptophan (PVA-pTR) binary blending adsorption fiber membrane is prepared.
(1) Static adsorption antibody assay
The adsorption fiber membranes of examples 1, 3 and 4 and comparative examples 1 and 2 were respectively taken as 1.0. 1.0G, plasma of a patient suffering from rheumatoid arthritis was added as 5.0 mL, and the adsorption rate of the adsorption fiber membrane to IgG, igM, igA was calculated by shaking the shaker to adsorb 2h (37 ℃, 100.+ -. 10 rpm) and detecting immunoglobulin G (IgG), immunoglobulin A (IgA) and immunoglobulin M (IgM) in the plasma.
TABLE 1 antibody adsorption test results
The adsorption test results are shown in table 1, and blending different polyaromatic amino acids slightly affects the adsorption effect of the antibody IgG, igM, igA. The adsorption results of the antibodies by the adsorption fiber membranes in comparative example 1, comparative example 1 and comparative example 2 can be seen: when the poly aromatic amino acid or lipoteichoic acid is not added to the blend solution, the adsorption effect of the adsorption fiber membrane on the immunoglobulin G (IgG), the immunoglobulin a (IgA) and the immunoglobulin M (IgM) is remarkably reduced, so that it can be inferred that the adsorption effect of the adsorption fiber membrane on the antibody can be synergistically improved by the lipoteichoic acid and the poly aromatic amino acid.
(2) Leukocyte adsorption assay
The adsorption fiber membranes of examples 1, 2, 3 and 4 and comparative examples 1 and 2 were taken as 1.0. 1.0g, fresh blood of volunteers anticoagulated with 5.0 mL heparin was added, and the adsorption was performed by shaking the shaker for adsorption 2h (37 ℃, 100.+ -. 10 rpm), and the white blood cell count was measured, and the adsorption rate of white blood cells by the adsorption fiber membrane was calculated.
TABLE 2 leukocyte adsorption test results
The adsorption test results are shown in Table 1, wherein the adsorption rate to leukocytes is slightly higher in example 2 than in examples 1 and 3, since the lipoteichoic acid content is higher in example 2 than in examples 1, 3 and 4. In addition, the adsorption effect of the adsorption fiber membrane on leukocytes in comparative example 1 and comparative examples 1 and 2 can be seen: when the adsorption fiber membrane is not added with lipoteichoic acid, the adsorption rate of the adsorption fiber membrane to the leucocytes is obviously reduced.
(3) Hemolysis test
The hemolysis rates of the adsorption fiber membranes of examples 1, 2, 3 and 4 were examined according to the hemolysis test method defined in GB/T14233.2. The test results are shown in Table 3.
TABLE 3 adsorption fiber membrane hemolysis test
The hemolysis rate of the adsorption fiber membrane is lower than 5% and meets the standard GB/T16886.4, so that the adsorption fiber membrane has good blood compatibility.
(4) Lipoteichoic acid abscission test
The purified examples 1, 2, 3 and 4, adsorption fiber membranes 1.0 and g are respectively put into a sterilized paper-plastic bag for sealing and sterilized by ethylene oxide, after sterilization, the materials are taken out, put into a conical flask, added with 10 ml sterile filtered bovine serum, oscillated for 2 hours at 37 ℃, and the supernatant is sucked and detected by lipoteichoic acid ELISA kit (the detection range is 0.4-500 EU/ml) according to the description operation, and the steps are performed for 3 times. The glass instrument used in the test is subjected to dry heat sterilization treatment in advance, and the whole test process is carried out in an ultra-clean workbench. The test results are shown in Table 4.
TABLE 4 adsorption fiber membrane lipoteichoic acid abscission test
The results in Table 4 show that the lipoteichoic acid falling-off amount of the supernatant is lower than 0.4 EU/ml under the simulated clinical use environment, and the adsorption fiber membrane has good stability, so that the application of the adsorption fiber membrane in blood purification meets the safety requirement.
The present embodiment is merely illustrative of the present application and is not limiting of the present application, and those skilled in the art, after having read the present specification, may make modifications to the present embodiment without creative contribution as necessary, but are protected by patent laws within the scope of the claims of the present application.
Claims (10)
1. An adsorption fiber membrane for removing leucocytes and pathogenic antibodies, which is characterized in that: preparing a fiber membrane from the blending solution through an electrostatic spinning technology, and then crosslinking and purifying the fiber membrane to obtain the fiber membrane; wherein the blend solution comprises a fibrous membrane matrix with good biocompatibility, lipoteichoic acid and polyaromatic amino acid, and the mass ratio of the fibrous membrane matrix, the lipoteichoic acid and the polyaromatic amino acid is (1-100): (1-10):1.
2. An absorbent fibrous membrane for removing leukocytes and pathogenic antibodies according to claim 1, wherein: the mass ratio of the fibrous membrane matrix to lipoteichoic acid to the poly aromatic amino acid is (4-20): (1-2):1.
3. An absorbent fibrous membrane for removing leukocytes and pathogenic antibodies according to claim 1, wherein: the fibrous membrane matrix is polyvinyl alcohol, the average molecular weight of the polyvinyl alcohol is 15-20 ten thousand, and the alcoholysis degree is 88-98%.
4. An absorbent fibrous membrane for removing leukocytes and pathogenic antibodies according to claim 1, wherein: the lipoteichoic acid is obtained by inactivating, crushing, extracting, purifying and freeze-drying any one or more strains of lactobacillus, staphylococcus aureus, streptococcus and bacillus subtilis.
5. An absorbent fibrous membrane for removing leukocytes and pathogenic antibodies according to claim 1, wherein: the average molecular weight of the poly-aromatic amino acid is 1000-20000.
6. An absorbent fibrous membrane for removing leukocytes and pathogenic antibodies according to claim 1, wherein: the polyaromatic amino acid is selected from phenylalanine and derivatives thereof, or polytyrosine and derivatives thereof, or polytryptone and derivatives thereof.
7. An absorbent fibrous membrane for removing leukocytes and pathogenic antibodies according to claim 1, wherein: the fiber diameter of the fiber membrane is 0.1-50 mu m, and the thickness of the fiber membrane is 100-1000 mu m.
8. A method for preparing an adsorption fiber membrane for removing white blood cells and pathogenic antibodies according to any one of claims 3 to 7, characterized in that: the method comprises the following steps:
s1, preparing a blending solution by adopting polyvinyl alcohol, lipoteichoic acid, poly aromatic amino acid and water;
s2, spinning the blending solution by adopting an electrostatic spinning technology, and then collecting and drying to prepare a fiber membrane;
s3, swelling the fiber membrane, and then adding glutaraldehyde solution into the swelled fiber membrane to perform heating crosslinking under alkaline conditions to obtain a crosslinked fiber membrane;
s4, purifying the crosslinked fiber membrane to obtain the adsorption fiber membrane.
9. The method for preparing the adsorption fiber membrane for removing white blood cells and pathogenic antibodies according to claim 8, wherein the method comprises the following steps: s2, spinning by adopting a needleless electrostatic spinning technology, wherein the rotating speed of a rotating cylinder is 3-6r/min, the collecting distance is 15-30cm, and the applied voltage is 20-60kV in the spinning process.
10. The method for preparing the adsorption fiber membrane for removing white blood cells and pathogenic antibodies according to claim 8, wherein the method comprises the following steps: and S3, controlling the crosslinking temperature in the S3 at 45-55 ℃ and the crosslinking time at 2-6h.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1518870A1 (en) * | 2003-09-17 | 2005-03-30 | Gambro Lundia AB | Separating material |
| CN109078619A (en) * | 2018-08-16 | 2018-12-25 | 重庆郑博生物科技有限公司 | A kind of adsorbent and purposes of wide spectrum absorption pyemia virulence factor |
| CN110913883A (en) * | 2017-06-13 | 2020-03-24 | 综合生物治疗疫苗公司 | Immunogenic composition comprising Staphylococcus aureus leukocidin LUKA and LUKB derived polypeptides |
| CN113274988A (en) * | 2021-04-30 | 2021-08-20 | 佛山市博新生物科技有限公司 | Adsorbing material for blood purification and preparation method thereof |
| CN114687072A (en) * | 2022-03-29 | 2022-07-01 | 北京服装学院 | Leukocyte-removing antibacterial composite nanofiber membrane as well as preparation method and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US20200360513A1 (en) * | 2019-05-17 | 2020-11-19 | Sila Nanotechnologies Inc. | Nanofiber compositions for a vaccine adjuvant, porous scaffold or porous membrane |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1518870A1 (en) * | 2003-09-17 | 2005-03-30 | Gambro Lundia AB | Separating material |
| CN110913883A (en) * | 2017-06-13 | 2020-03-24 | 综合生物治疗疫苗公司 | Immunogenic composition comprising Staphylococcus aureus leukocidin LUKA and LUKB derived polypeptides |
| CN109078619A (en) * | 2018-08-16 | 2018-12-25 | 重庆郑博生物科技有限公司 | A kind of adsorbent and purposes of wide spectrum absorption pyemia virulence factor |
| CN113274988A (en) * | 2021-04-30 | 2021-08-20 | 佛山市博新生物科技有限公司 | Adsorbing material for blood purification and preparation method thereof |
| CN114687072A (en) * | 2022-03-29 | 2022-07-01 | 北京服装学院 | Leukocyte-removing antibacterial composite nanofiber membrane as well as preparation method and application thereof |
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