CN113117066A - Preparation method of chitosan nanoparticle vaccine for avian influenza - Google Patents
Preparation method of chitosan nanoparticle vaccine for avian influenza Download PDFInfo
- Publication number
- CN113117066A CN113117066A CN202110302260.4A CN202110302260A CN113117066A CN 113117066 A CN113117066 A CN 113117066A CN 202110302260 A CN202110302260 A CN 202110302260A CN 113117066 A CN113117066 A CN 113117066A
- Authority
- CN
- China
- Prior art keywords
- chitosan
- avian influenza
- solution
- virus
- preparing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/5115—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/513—Organic macromolecular compounds; Dendrimers
- A61K9/5161—Polysaccharides, e.g. alginate, chitosan, cellulose derivatives; Cyclodextrin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
- A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/525—Virus
- A61K2039/5252—Virus inactivated (killed)
-
- 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
- C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
- C12N2760/00011—Details
- C12N2760/16011—Orthomyxoviridae
- C12N2760/16111—Influenzavirus A, i.e. influenza A virus
- C12N2760/16134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Abstract
The invention relates to the technical field of biological products, in particular to a preparation method of chitosan nanoparticle vaccine for avian influenza, which comprises the following steps: s1, preparing a chitosan solution and a sodium tripolyphosphate solution; s2, preparing inactivated virus liquid; s3, dropwise adding the inactivated virus liquid into the chitosan solution to obtain a virus/chitosan mixed solution; and adding the sodium tripolyphosphate solution into the virus/chitosan mixed solution at a flow rate of 0.1-0.5 mL/min under the condition of continuously stirring at a rotating speed of 800-1500 rpm/min. The invention has great potential in the aspect of preventing H9N2 avian influenza, is expected to be used as a novel nano vaccine to be applied to clinic, and has good vaccine development potential. In addition, the method provided by the invention can be applied to a wider range, different virus types can be selected, or a certain virus is subjected to genetic engineering modification, so that the antigen region of the virus is utilized for preparing the vaccine.
Description
Technical Field
The invention relates to the technical field of biological products, in particular to a preparation method of a chitosan nanoparticle vaccine for avian influenza.
Background
Avian influenza is a high-grade disease of poultry caused by influenza A virus, and takes respiratory symptoms such as mental depression, anorexia, cough, sneeze and the like as main clinical symptoms. At present, avian influenza caused by H9N2 is endemic in China, although the mortality is low, infected laying hens are easy to have abnormal oviduct function, lay faded eggs, malformed eggs, preserved eggs and soft-shell eggs, the laying rate of a chicken group is reduced by 30-80%, the duration of the disease course is long and can reach more than one month, and serious economic loss is caused. China also happens successively in recent years, which causes huge economic loss for the poultry industry. Vaccination remains the main means for preventing and controlling H9N2 avian influenza, while nanoparticles refer to tiny particles with a particle diameter size of 1-1000 nm. Due to the physical and biological characteristics, the method can be widely applied to the aspects of food, chemical industry, detection, textile and machinery, biomedicine and the like. The nano particles can be designed into a natural virus form (virus-like particles) through precision processing, and can be loaded with multiple antigens simultaneously, so that the nano particles have wide application prospects in the aspect of vaccines.
The traditional avian influenza vaccines are generally divided into three types, namely inactivated vaccines, attenuated live vaccines and genetic engineering vaccines. The traditional vaccine has certain limitations, such as the situation that attenuated live vaccine may have virulence reversion, the duration of immunity generated by inactivated vaccine is short, plasmid DNA vaccine may be integrated into the chromosome of the immunized animal, and the immune stress reaction is severe.
Disclosure of Invention
The invention aims to solve the problems that attenuated live vaccines in the prior art are likely to have a virulence reversion phenomenon, inactivated vaccines are short in duration of immunity generation, plasmid DNA vaccines are likely to be integrated into chromosomes of immunized animals, immune stress reaction is severe and the like, and provides a preparation method of chitosan nanoparticle vaccines for avian influenza.
The purpose of the invention is realized by the following technical scheme:
a preparation method of chitosan nanoparticle vaccine for avian influenza comprises the following steps:
s1, preparing a chitosan solution and a sodium tripolyphosphate solution;
s2, preparing inactivated virus liquid;
s3, dropwise adding the inactivated virus liquid into the chitosan solution to obtain a virus/chitosan mixed solution; adding a sodium tripolyphosphate solution into the virus/chitosan mixed solution at a flow rate of 0.1-0.5 mL/min under the condition of continuously stirring at a rotating speed of 800-1500 rpm/min;
s4, centrifuging the mixed solution obtained in the step S3 at a rotating speed of 8000-13000 g/min for 20-50 min to obtain chitosan-avian influenza nanoparticles at the temperature of 1-8 ℃; dispersing the obtained chitosan-avian influenza nano-particles in a phosphoric acid buffer solution.
Preparing a chitosan solution: the chitosan solid particles were dissolved in 2% aqueous acetic acid until the solution was clear and stirred with a magnetic stirrer until complete dissolution. After dissolution, the chitosan solution was diluted with deionized water and the pH of the solution was adjusted to 5.4 with NaOH. The solution was then filtered through a 0.22 μm filter to give a chitosan solution for use in the experiment.
Preparing a sodium tripolyphosphate solution: the solid particles of sodium tripolyphosphate are dissolved in deionized water and stirred continuously with a magnetic stirrer until completely dissolved. The solution was filtered through a 0.22 μm filter to obtain a sodium tripolyphosphate solution for the experiment.
According to the principle of ionic crosslinking, the nanoparticles are formed by intramolecular and intermolecular crosslinking between positively charged chitosan and negatively charged sodium tripolyphosphate.
Preferably, in the step S1, the concentration of the chitosan solution is 0.5% to 2% (w/v); the concentration of the sodium tripolyphosphate is 0.5-1.5% (w/v).
Preferably, in the step S3, the concentration of the inactivated virus is 0.05-0.15% (mg/v), and the volume ratio of the inactivated virus to the chitosan solution is 1 (1-3).
Preferably, in the step S3, the volume ratio of the virus/chitosan mixed solution to the sodium tripolyphosphate solution is (4-6): 1.
Preferably, in the step S4, the obtained chitosan-avian influenza nanoparticles are dispersed in a phosphoric acid buffer solution by using ultrasound for 5-15 min.
Preferably, the inactivated virus is H9N2 avian influenza virus.
The avian influenza chitosan nanoparticle vaccine is obtained by the preparation method of the avian influenza chitosan nanoparticle vaccine.
Compared with the prior art, the invention has the following technical effects:
the preparation method of the chitosan nanoparticle vaccine for avian influenza disclosed by the invention has the characteristics of high safety and convenience in preparation, is simple and rapid in preparation, high in efficiency and time-saving, and is beneficial to popularization and application, and meanwhile, the vaccine prepared by the invention has strong specificity and small toxic and side effects, has great potential in the aspect of preventing H9N2 avian influenza, is expected to be applied to clinic as a novel nano vaccine, and has good vaccine development potential. In addition, the method provided by the invention can be applied to a wider range, different virus types can be selected, or a certain virus is subjected to genetic engineering modification, so that the antigen region of the virus is utilized for preparing the vaccine.
Drawings
FIG. 1 size and size distribution diagram of chitosan nanoparticle vaccine of example 1 of the present invention;
FIG. 2 is an electron micrograph showing the morphology of the chitosan nanoparticle vaccine according to example 1 of the present invention;
FIG. 3 is a graph showing HI analysis results of antibody titers of chitosan nanoparticle vaccines at different concentration ratios for eliciting humoral immune response of SPF white-feather chickens;
FIG. 4 is a graph showing the results of analyzing the proliferation of SPF chicken lymphocytes induced by chitosan nanoparticle vaccine with different concentration ratios.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Unless otherwise specified, the devices used in the present examples, comparative examples and experimental examples were all conventional experimental devices, the materials and reagents used were commercially available without specific reference, and the experimental methods without specific reference were also conventional experimental methods.
Example 1
A preparation method of chitosan nanoparticle vaccine for avian influenza comprises the following steps:
s1, preparing a chitosan solution with the concentration of 1% (w/v) and a sodium tripolyphosphate solution with the concentration of 1% (w/v);
s2, preparing inactivated virus liquid; utilizing SPF chick embryo to amplify virus, collecting allantoic fluid, inactivating the harvested virus allantoic fluid by using formaldehyde to obtain a final concentration of 0.1%, reacting in a shaker at 37 ℃ and 400rpm/min for 48h, and filtering by using a 0.22 mu m filter;
s3, dropwise adding 2.5mL of inactivated virus liquid into 5.0mL of chitosan solution to obtain a virus/chitosan mixed solution; adding a sodium tripolyphosphate solution into the virus/chitosan mixed solution at a flow rate of 1/6mL/min under the condition of continuously stirring at a rotation speed of 1000rpm/min, wherein the volume ratio of the chitosan solution to the sodium tripolyphosphate solution is 5: 1;
s4, centrifuging the mixed solution obtained in the step S3 at the rotating speed of 10000g/min for 30min at the temperature of 4 ℃ to obtain chitosan-avian influenza nano particles; the obtained chitosan-avian influenza nanoparticles were dispersed in phosphoric acid buffer solution and sonicated for 10 mim.
The avian influenza chitosan nanoparticle vaccine is obtained by the preparation method of the avian influenza chitosan nanoparticle vaccine.
Example 2
A preparation method of chitosan nanoparticle vaccine for avian influenza comprises the following steps:
s1, preparing a chitosan solution with the concentration of 0.5% (w/v) and a sodium tripolyphosphate solution with the concentration of 0.5% (w/v);
s2, preparing inactivated virus liquid; utilizing SPF chick embryo to amplify virus, collecting allantoic fluid, inactivating the harvested virus allantoic fluid by using formaldehyde to obtain a final concentration of 0.1%, reacting in a shaker at 37 ℃ and 400rpm/min for 48h, and filtering by using a 0.22 mu m filter;
s3, dropwise adding 2.5mL of inactivated virus liquid into 2.5mL of chitosan solution to obtain a virus/chitosan mixed solution; adding a sodium tripolyphosphate solution into the virus/chitosan mixed solution at a flow rate of 0.2mL/min and under the condition of continuously stirring at a rotating speed of 1000rpm/min, wherein the volume ratio of the chitosan solution to the sodium tripolyphosphate solution is 4: 1;
s4, centrifuging the mixed solution obtained in the step S3 at the rotating speed of 13000g/min for 20min to obtain chitosan-avian influenza nano particles at the temperature of 1 ℃; the obtained chitosan-avian influenza nanoparticles were dispersed in phosphoric acid buffer solution and sonicated for 10 mim.
The avian influenza chitosan nanoparticle vaccine is obtained by the preparation method of the avian influenza chitosan nanoparticle vaccine.
Example 3
A preparation method of chitosan nanoparticle vaccine for avian influenza comprises the following steps:
s1, preparing a chitosan solution with the concentration of 2% (w/v) and a sodium tripolyphosphate solution with the concentration of 1.5% (w/v);
s2, preparing inactivated virus liquid; utilizing SPF chick embryo to amplify virus, collecting allantoic fluid, inactivating the harvested virus allantoic fluid by using formaldehyde to obtain a final concentration of 0.1%, reacting in a shaker at 37 ℃ and 400rpm/min for 48h, and filtering by using a 0.22 mu m filter;
s3, dropwise adding 2.5mL of inactivated virus liquid into 7.5mL of chitosan solution to obtain a virus/chitosan mixed solution; adding a sodium tripolyphosphate solution into the virus/chitosan mixed solution at a flow rate of 0.5mL/min and under the condition of continuously stirring at a rotating speed of 800rpm/min, wherein the volume ratio of the chitosan solution to the sodium tripolyphosphate solution is 6: 1;
s4, centrifuging the mixed solution obtained in the step S3 at the rotating speed of 8000g/min for 50min to obtain chitosan-avian influenza nano particles at the temperature of 8 ℃; the obtained chitosan-avian influenza nanoparticles were dispersed in phosphoric acid buffer solution and sonicated for 10 mim.
The avian influenza chitosan nanoparticle vaccine is obtained by the preparation method of the avian influenza chitosan nanoparticle vaccine.
Comparative example 1
A preparation method of chitosan nanoparticle vaccine for avian influenza comprises the following steps:
s1, preparing a chitosan solution with the concentration of 1% (w/v) and a sodium tripolyphosphate solution with the concentration of 1% (w/v);
s2, preparing inactivated virus liquid; utilizing SPF chick embryo to amplify virus, collecting allantoic fluid, inactivating the harvested virus allantoic fluid by using formaldehyde to obtain a final concentration of 0.1%, reacting in a shaker at 37 ℃ and 400rpm/min for 48h, and filtering by using a 0.22 mu m filter;
s3, dropwise adding 2.5mL of inactivated virus liquid into 5.0mL of chitosan solution to obtain a virus/chitosan mixed solution; adding the sodium tripolyphosphate solution into the virus/chitosan mixed solution at the flow rate of 1/6mL/min under the condition of continuously stirring at the rotating speed of 1000rpm/min, wherein the volume ratio of the chitosan solution to the sodium tripolyphosphate solution is 8: 1;
s4, centrifuging the mixed solution obtained in the step S3 at the rotating speed of 10000g/min for 30min at the temperature of 4 ℃ to obtain chitosan-avian influenza nano particles; the obtained chitosan-avian influenza nanoparticles were dispersed in phosphoric acid buffer solution and sonicated for 10 mim.
The avian influenza chitosan nanoparticle vaccine is obtained by the preparation method of the avian influenza chitosan nanoparticle vaccine.
Compared with example 1, the volume ratio of the virus/chitosan mixed solution to the sodium tripolyphosphate solution in the comparative example is different.
Comparative example 2
A preparation method of chitosan nanoparticle vaccine for avian influenza comprises the following steps:
s1, preparing a chitosan solution with the concentration of 1% (w/v) and a sodium tripolyphosphate solution with the concentration of 1% (w/v);
s2, preparing inactivated virus liquid; utilizing SPF chick embryo to amplify virus, collecting allantoic fluid, inactivating the harvested virus allantoic fluid by using formaldehyde to obtain a final concentration of 0.1%, reacting in a shaker at 37 ℃ and 400rpm/min for 48h, and filtering by using a 0.22 mu m filter;
s3, dropwise adding 2.5mL of inactivated virus liquid into 5.0mL of chitosan solution to obtain a virus/chitosan mixed solution; adding the sodium tripolyphosphate solution into the virus/chitosan mixed solution at the flow rate of 1/6mL/min under the condition of continuously stirring at the rotating speed of 1000rpm/min, wherein the volume ratio of the chitosan solution to the sodium tripolyphosphate solution is 2: 1;
s4, centrifuging the mixed solution obtained in the step S3 at the rotating speed of 10000g/min for 30min at the temperature of 4 ℃ to obtain chitosan-avian influenza nano particles; the obtained chitosan-avian influenza nanoparticles were dispersed in phosphoric acid buffer solution and sonicated for 10 mim.
The avian influenza chitosan nanoparticle vaccine is obtained by the preparation method of the avian influenza chitosan nanoparticle vaccine.
Compared with example 1, the volume ratio of the virus/chitosan mixed solution to the sodium tripolyphosphate solution in the comparative example is different.
Experimental example 1
Size and distribution of particles: as shown in fig. 1, the particle size and particle size distribution of the vaccine were measured using a BT-90 nm laser particle size distribution instrument of the dandongbett instruments ltd. The abscissa of the graph is the size of the nanoparticle vaccine and the ordinate is the percentage of the nanoparticle vaccine in a certain size. Particle size range is derived from the figure: the smaller the nano size is, the smaller the size is, the 300-900nm is helpful for helping the vaccine enter cells and enhancing immunity. As shown in fig. 2, transmission electron microscopy FEI/Talos L120c measured the characterization of avian influenza nanoparticle vaccine. In the figure, a large circle is an ultrathin carbon net, and a small circle (black) is the morphological characterization of the avian influenza nanoparticle vaccine under an electron microscope.
Experimental example 2
60 SPF white-feathered chickens, 4 weeks old, were randomly divided into six groups of 10 chickens each. The chickens were immunized subcutaneously (S/C) with 0.5mL of each of the chitosan nanoparticle vaccines of the example or comparative example groups, one of which served as a negative control group. The chickens were immunized a second time 3 weeks after the first immunization and the immunization schedule is shown in table 1.
TABLE 1 immunization protocol
| Group of | First immunization | Second immunization |
| Example 1 | S/C 0.5mL | S/C 0.5mL |
| Example 2 | S/C 0.5mL | S/C 0.5mL |
| Example 3 | S/C 0.5mL | S/C 0.5mL |
| Comparative example 1 | S/C 0.5mL | S/C 0.5mL |
| Comparative example 2 | S/C 0.5mL | S/C 0.5mL |
| Control group | NO | NO |
Determination of specific antibodies (HI experiment): according to the Hemagglutination Inhibition (HI) assay published in the OIE manual. The HI experiment is performed on serum samples collected from an immunization group with a test avian influenza vaccine and a control group to detect the presence of antibodies against H9N2 in the samples. Measurements were taken weekly within 3 weeks after the first immunization and every 2 weeks after the second immunization. A disposable vacuum collection tube was used to collect 3-5mL of blood, and then serum was collected for HI experiments. The test sera were diluted in 2-fold serial dilutions and mixed with equal volumes of avian influenza antigen (Ags) containing 4 HA units each. An equal volume of 0.5% turkey Red Blood Cells (RBC) was then added to the mixture and the diluent examined for hemagglutination inhibition. The serum antibody titer is the reciprocal of the highest dilution that completely inhibits hemagglutination of avian influenza virus. The results are shown in FIG. 3. The abscissa of the graph is the number of days after the first immunization, and the ordinate is the HI value (antibody titer value). Comparison with other experimental groups: example 1 has the highest value, elicits stronger humoral immunity, and induces the body to produce stronger antibody titers.
Lymphocyte isolation and proliferation assay: (from each group of 3 selected chicken) using XTT cell proliferation assay kit for lymphocyte proliferation assay. On day 14 after the first immunization, blood was collected from each chicken by winged venipuncture with heparin (20 μm/mL) for lymphocyte proliferation assay. The separation of lymphocytes from peripheral blood of chicken was carried out by using a lymphocyte separation medium. The results of lymphocyte proliferation are shown in FIG. 4. The abscissa in the figure is the group, each group comprising four values, and the ordinate is the OD value. Comparison with other experimental groups: the higher the value of example 1, the stronger cellular immunity is elicited, and stimulation of lymphocytes and the like produces a stronger immune response.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (7)
1. A preparation method of chitosan nanoparticle vaccine for avian influenza is characterized by comprising the following steps:
s1, preparing a chitosan solution and a sodium tripolyphosphate solution;
s2, preparing inactivated virus liquid;
s3, dropwise adding the inactivated virus liquid into the chitosan solution to obtain a virus/chitosan mixed solution; adding a sodium tripolyphosphate solution into the virus/chitosan mixed solution at a flow rate of 0.1-0.5 mL/min under the condition of continuously stirring at a rotating speed of 800-1500 rpm/min;
s4, centrifuging the mixed solution obtained in the step S3 at a rotating speed of 8000-13000 g/min for 20-50 min to obtain chitosan-avian influenza nanoparticles at the temperature of 1-8 ℃; dispersing the obtained chitosan-avian influenza nano-particles in a phosphoric acid buffer solution.
2. The method for preparing chitosan nanoparticle vaccine for avian influenza according to claim 1, wherein in the step S1, the concentration of chitosan solution is 0.5% -2% (w/v); the concentration of the sodium tripolyphosphate is 0.5-1.5% (w/v).
3. The method for preparing chitosan nanoparticle vaccine for avian influenza according to claim 1, wherein in the step S3, the concentration of the inactivated virus is 0.05% -0.15% (mg/v), and the volume ratio of the inactivated virus to the chitosan solution is 1 (1-3).
4. The method for preparing chitosan nanoparticle vaccine for avian influenza according to claim 1, wherein in the step S3, the volume ratio of the virus/chitosan mixed solution to the sodium tripolyphosphate solution is (4-6): 1.
5. The method for preparing chitosan nanoparticle vaccine for avian influenza according to claim 1, wherein in step S4, the obtained chitosan-avian influenza nanoparticles are dispersed in phosphoric acid buffer solution by using ultrasound for 5-15 min.
6. The method for preparing chitosan nanoparticle vaccine for avian influenza according to claim 1, wherein the inactivated virus is H9N2 avian influenza virus.
7. The chitosan nanoparticle vaccine for avian influenza obtained by the method for preparing the chitosan nanoparticle vaccine for avian influenza of any one of claims 1 to 5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110302260.4A CN113117066A (en) | 2021-03-22 | 2021-03-22 | Preparation method of chitosan nanoparticle vaccine for avian influenza |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110302260.4A CN113117066A (en) | 2021-03-22 | 2021-03-22 | Preparation method of chitosan nanoparticle vaccine for avian influenza |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113117066A true CN113117066A (en) | 2021-07-16 |
Family
ID=76773704
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110302260.4A Pending CN113117066A (en) | 2021-03-22 | 2021-03-22 | Preparation method of chitosan nanoparticle vaccine for avian influenza |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113117066A (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102327228A (en) * | 2011-09-26 | 2012-01-25 | 黑龙江大学 | Preparation method of chitosan newcastle disease vaccine nanoparticles |
| CN102586194A (en) * | 2011-12-01 | 2012-07-18 | 哈药集团生物疫苗有限公司 | H9 subtype avian influenza virus Vero cell adapted strain and application thereof |
| CN102580083A (en) * | 2012-04-06 | 2012-07-18 | 黑龙江大学 | Method for preparing nanoparticle oil adjuvant vaccine |
| CN102973936A (en) * | 2011-12-14 | 2013-03-20 | 普莱柯生物工程股份有限公司 | Application of chitosan in avian vaccine composition preparation |
| CN103877570A (en) * | 2014-02-19 | 2014-06-25 | 北京中联康生物科技有限公司 | Rabies live vaccine and method for preparing rabies live vaccine for oral administration |
| CN106039303A (en) * | 2016-06-20 | 2016-10-26 | 华南农业大学 | H5 subtype avian influenza DNA vaccine and preparation method thereof |
| CN108310374A (en) * | 2017-12-30 | 2018-07-24 | 华南农业大学 | A kind of preparation method of avian influenza virus inactivated vaccine |
| CN108379239A (en) * | 2018-02-09 | 2018-08-10 | 中山大学 | A kind of nano vaccine and its preparation method and application of the controllable antigen rate of release based on chitosan |
| US20190125661A1 (en) * | 2016-04-12 | 2019-05-02 | Capsular Technologies Pty Ltd | Injectable composition for delivery of a biologically active agent |
-
2021
- 2021-03-22 CN CN202110302260.4A patent/CN113117066A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102327228A (en) * | 2011-09-26 | 2012-01-25 | 黑龙江大学 | Preparation method of chitosan newcastle disease vaccine nanoparticles |
| CN102586194A (en) * | 2011-12-01 | 2012-07-18 | 哈药集团生物疫苗有限公司 | H9 subtype avian influenza virus Vero cell adapted strain and application thereof |
| CN102973936A (en) * | 2011-12-14 | 2013-03-20 | 普莱柯生物工程股份有限公司 | Application of chitosan in avian vaccine composition preparation |
| CN102580083A (en) * | 2012-04-06 | 2012-07-18 | 黑龙江大学 | Method for preparing nanoparticle oil adjuvant vaccine |
| CN103877570A (en) * | 2014-02-19 | 2014-06-25 | 北京中联康生物科技有限公司 | Rabies live vaccine and method for preparing rabies live vaccine for oral administration |
| US20190125661A1 (en) * | 2016-04-12 | 2019-05-02 | Capsular Technologies Pty Ltd | Injectable composition for delivery of a biologically active agent |
| CN106039303A (en) * | 2016-06-20 | 2016-10-26 | 华南农业大学 | H5 subtype avian influenza DNA vaccine and preparation method thereof |
| CN108310374A (en) * | 2017-12-30 | 2018-07-24 | 华南农业大学 | A kind of preparation method of avian influenza virus inactivated vaccine |
| CN108379239A (en) * | 2018-02-09 | 2018-08-10 | 中山大学 | A kind of nano vaccine and its preparation method and application of the controllable antigen rate of release based on chitosan |
Non-Patent Citations (3)
| Title |
|---|
| IRAJ KHALILI 等: "Evaluation of Immune Response Against Inactivated Avian Influenza (H9N2) Vaccine, by Using Chitosan Nanoparticles", 《JUNDISHAPUR J MICROBIOL》 * |
| SAMAH H MOHAMED 等: "Preparation and immunological evaluation of inactivated avian influenza virus vaccine encapsulated in chitosan nanoparticles", 《BIOLOGICALS》 * |
| 潘亮奇 等: "禽流感纳米颗粒疫苗的研究进展", 《中国兽医科学》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA2250078C (en) | Processes for the replication of influenza viruses in cell culture, and the influenza viruses obtainable by the process | |
| CA2197683C (en) | Method for preparing an influenza virus, antigens obtained and applications thereof | |
| CN105412921B (en) | A kind of I group of 4 type aviadenovirus vaccine | |
| CN101491672A (en) | Chicken new castle disease-bird influenza dual oil-emulsion inactivated vaccine and preparation method thereof | |
| CN106924727B (en) | Preparation method of avian influenza virus H9 subtype inactivated vaccine | |
| CN102805864B (en) | Newcastle disease and H9N2 subtype avian influenza bivalent inactivated vaccine and preparation method thereof | |
| JP3950500B2 (en) | Iridovirus infectious disease vaccine and diagnostic agent for fish and production method thereof | |
| CN107050448B (en) | Preparation method of avian influenza virus and avian adenovirus bivalent inactivated vaccine | |
| CN108721615A (en) | A kind of method and its vaccine preparing duck tembusu virus inactivated vaccine | |
| CN104730259A (en) | Method for detecting hemagglutination inhibition antibody of chicken infectious bronchitis | |
| CN102327228A (en) | Preparation method of chitosan newcastle disease vaccine nanoparticles | |
| CN104212771A (en) | Avian influenza virus proliferation method and avian influenza vaccine prepared by use of avian influenza virus proliferation method | |
| CN109097340A (en) | A kind of aviadenovirus, a kind of quadruple vaccine and preparation method thereof | |
| CN113117066A (en) | Preparation method of chitosan nanoparticle vaccine for avian influenza | |
| CN105664150A (en) | Newcastle disease virus, avian influenza virus and avian adenovirus triple inactivated vaccine | |
| CN104164408A (en) | Newcastle disease, infectious bronchitis and avian influenza resisting vaccine composition and preparation | |
| Tevethia et al. | Antigenic characterization of infectious bronchitis virus | |
| CN1843507B (en) | Mumps virus ingredient vaccine for human, and its preparation method and uses | |
| CN107287168A (en) | A kind of NDV saves method and its application | |
| CN105886477A (en) | Virus strain capable of being used for preparing VII type newcastle disease vaccine and encoding gene thereof | |
| CN113957007B (en) | Inactivated vaccine for mycoplasma synoviae | |
| KR20170030635A (en) | Vaccine containing virus-like particles | |
| CN103789272A (en) | H9 subtype avian influenza virus isolate and vaccine composition prepared thereby | |
| CN106075424B (en) | One avian influenza virus vaccine | |
| CN104667270A (en) | Avian infectious bronchitis viral antigen and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |