CN111184871A - Heat-resistant protective agent for newcastle disease and bronchitis combined live vaccine and preparation process thereof - Google Patents
Heat-resistant protective agent for newcastle disease and bronchitis combined live vaccine and preparation process thereof Download PDFInfo
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- CN111184871A CN111184871A CN202010206099.6A CN202010206099A CN111184871A CN 111184871 A CN111184871 A CN 111184871A CN 202010206099 A CN202010206099 A CN 202010206099A CN 111184871 A CN111184871 A CN 111184871A
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- 239000003223 protective agent Substances 0.000 title claims abstract description 48
- 229960005486 vaccine Drugs 0.000 title claims abstract description 39
- 208000010359 Newcastle Disease Diseases 0.000 title claims abstract description 28
- 206010006451 bronchitis Diseases 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- 239000000243 solution Substances 0.000 claims abstract description 53
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 30
- 239000008215 water for injection Substances 0.000 claims abstract description 28
- MKJXYGKVIBWPFZ-UHFFFAOYSA-L calcium lactate Chemical compound [Ca+2].CC(O)C([O-])=O.CC(O)C([O-])=O MKJXYGKVIBWPFZ-UHFFFAOYSA-L 0.000 claims abstract description 20
- 229960002401 calcium lactate Drugs 0.000 claims abstract description 20
- 235000011086 calcium lactate Nutrition 0.000 claims abstract description 20
- 239000001527 calcium lactate Substances 0.000 claims abstract description 20
- UMGDCJDMYOKAJW-UHFFFAOYSA-N thiourea Chemical compound NC(N)=S UMGDCJDMYOKAJW-UHFFFAOYSA-N 0.000 claims abstract description 20
- 241000287828 Gallus gallus Species 0.000 claims abstract description 14
- 230000001502 supplementing effect Effects 0.000 claims abstract description 13
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 12
- 229930006000 Sucrose Natural products 0.000 claims abstract description 12
- 239000005720 sucrose Substances 0.000 claims abstract description 12
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims abstract description 10
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims abstract description 10
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims abstract description 10
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims abstract description 10
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Natural products NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims abstract description 10
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims abstract description 10
- 229940098773 bovine serum albumin Drugs 0.000 claims abstract description 10
- 229960001855 mannitol Drugs 0.000 claims abstract description 10
- 239000001267 polyvinylpyrrolidone Substances 0.000 claims abstract description 10
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 claims abstract description 10
- 229920000036 polyvinylpyrrolidone Polymers 0.000 claims abstract description 10
- 229940073490 sodium glutamate Drugs 0.000 claims abstract description 9
- 241000711450 Infectious bronchitis virus Species 0.000 claims abstract description 6
- 241000711404 Avian avulavirus 1 Species 0.000 claims abstract description 5
- 239000000427 antigen Substances 0.000 claims abstract description 4
- 102000036639 antigens Human genes 0.000 claims abstract description 4
- 108091007433 antigens Proteins 0.000 claims abstract description 4
- 238000001035 drying Methods 0.000 claims description 22
- 238000004108 freeze drying Methods 0.000 claims description 14
- 238000001914 filtration Methods 0.000 claims description 10
- 238000010438 heat treatment Methods 0.000 claims description 10
- 238000003756 stirring Methods 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 9
- 230000001954 sterilising effect Effects 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 5
- 238000007710 freezing Methods 0.000 claims description 4
- 230000008014 freezing Effects 0.000 claims description 4
- 238000004659 sterilization and disinfection Methods 0.000 claims description 3
- 238000002347 injection Methods 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 claims 1
- 241000700605 Viruses Species 0.000 abstract description 14
- 238000012360 testing method Methods 0.000 abstract description 9
- 210000003746 feather Anatomy 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 241001465754 Metazoa Species 0.000 abstract description 2
- 238000001514 detection method Methods 0.000 abstract 1
- 235000013330 chicken meat Nutrition 0.000 description 12
- 208000015181 infectious disease Diseases 0.000 description 8
- 230000002458 infectious effect Effects 0.000 description 8
- 238000011160 research Methods 0.000 description 5
- 238000003860 storage Methods 0.000 description 5
- 210000003837 chick embryo Anatomy 0.000 description 3
- 238000010998 test method Methods 0.000 description 3
- 208000010884 Bursa disease Diseases 0.000 description 1
- 201000006082 Chickenpox Diseases 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 208000006758 Marek Disease Diseases 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 206010046980 Varicella Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000009993 protective function Effects 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
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- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
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- 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
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
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- A61K47/20—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing sulfur, e.g. dimethyl sulfoxide [DMSO], docusate, sodium lauryl sulfate or aminosulfonic acids
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- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
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- C12N2760/00011—Details
- C12N2760/18011—Paramyxoviridae
- C12N2760/18111—Avulavirus, e.g. Newcastle disease virus
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- C12N2770/00011—Details
- C12N2770/20011—Coronaviridae
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Abstract
The invention relates to the technical field of production of vaccines for animals, in particular to a heat-resistant protective agent of a chicken newcastle disease and bronchitis bigeminal live vaccine and a preparation process thereof, wherein the heat-resistant protective agent consists of a solution A and a solution B according to the volume ratio of 4: 1; wherein the solution A comprises 20-30g of calcium lactate, 5-10g of thiourea, 15-20g of sucrose, 3-5g of trehalose, 5-10g of L-sodium glutamate and 10-20g of bovine serum albumin; supplementing water for injection to 800 ml; the solution B consists of the following components: 4-6g of polyvinylpyrrolidone and 3-5g of d-mannitol, and the volume is increased to 200mL by using water for injection; the protective agent is mixed with antigens of Newcastle disease and infectious bronchitis virus according to the proportion of 1: 1, subpackaged, freeze-dried, the virus content after being placed at 37 ℃ is compared with the virus content of effectiveness test, the reduction range of each feather of the virus content of Newcastle disease and infectious bronchitis virus is not more than 1 titer after 10 days, all detection results are qualified, the performance of the protective agent is stable, and the yield of the protective agent produced by the preparation process of the heat-resistant protective agent is high.
Description
Technical Field
The invention relates to the technical field of production of vaccines for animals, in particular to a heat-resistant protective agent of a chicken newcastle disease and bronchitis bigeminal live vaccine and a preparation process thereof.
Background
The freeze-drying protective agent and the preparation process thereof are a key technology in the production of live vaccines, and have great influence on the storage, transportation and use of the vaccines. At present, in domestic veterinary vaccines, live freeze-dried vaccines account for a considerable proportion, and domestic commonly used protective agents mainly comprise milk, sucrose, gelatin and the like, and the live freeze-dried vaccines have simple formula and poor protective function on the vaccines. The vaccine freeze-dried by the traditional protective agent usually requires that the product is kept below-15 ℃, which causes great inconvenience to manufacturers and users, is limited in long-term storage and transportation of the vaccine, and may cause inactivation and efficacy reduction of the vaccine due to improper storage of the vaccine, so that the aim of epidemic prevention cannot be achieved, and huge economic loss is caused.
Compared with the conventional freeze-drying protective agent, the heat-resistant freeze-drying protective agent has the characteristic of long high-temperature-resistant storage period, so that the heat-resistant protective agent has better performance than the conventional protective agent, the live vaccine can be stored at 2-8 ℃ in the valid period, and can be stored for dozens of days even at higher room temperature, and the storage, transportation and use of the live vaccine are more convenient and economical.
The research on the heat-resistant protective agent in the developed countries starts earlier, and at present, the heat-resistant protective agent is basically used in the freeze-dried live vaccines of the developed countries, but the research is not reported in detail. The development of the domestic veterinary biological product protective agent is relatively lagged, and in recent years, China also has some related research reports, mainly including the research on heat-resistant freeze-drying protective agents for chicken newcastle disease, chicken Marek's disease, chicken bursa disease, chicken infectious bronchitis, chicken pox, swine fever freeze-drying live vaccines and the like. The research on the heat-resistant freeze-drying protective agent of the newcastle disease and infectious bronchitis combined live vaccine usually uses 5% sucrose skim milk as the protective agent, the performance of the product is not stable enough, and the passing yield is low.
Disclosure of Invention
In order to solve the problems, the invention provides a heat-resistant protective agent of a chicken newcastle disease and bronchitis bivalent live vaccine and a preparation process thereof.
The technical problem to be solved by the invention is realized by adopting the following technical scheme:
a heat-resistant protective agent for chicken newcastle disease and bronchitis bigeminal live vaccine comprises solution A and solution B at a volume ratio of 4: 1; wherein the solution A consists of the following components: 20-30g of calcium lactate, 5-10g of thiourea, 15-20g of sucrose, 3-5g of trehalose, 5-10g of L-sodium glutamate and 10-20g of bovine serum albumin; supplementing water for injection to 800 ml; the solution B consists of the following components: 4-6g of polyvinylpyrrolidone and 3-5g of d-mannitol, and the volume is increased to 200mL by using water for injection.
Preferably, the solution A consists of the following components: 30g of calcium lactate, 10g of thiourea, 20g of sucrose, 4g of trehalose, 6g of L-sodium glutamate and 16g of bovine serum albumin; the injection water is supplemented to 800 ml.
Preferably, the liquid B consists of the following components: 5g of polyvinylpyrrolidone and 4g of d-mannitol, and the volume of the solution is made up to 200mL with water for injection.
A preparation process of a chicken Newcastle disease and bronchitis bivalent live vaccine heat-resistant protective agent comprises the following steps:
s1: firstly, heating 500ml of water for injection to boil, slowly adding calcium lactate, stirring until the calcium lactate is completely dissolved and transparent, and filtering by qualitative filter paper; dissolving thiourea, sucrose, trehalose, L-sodium glutamate and bovine serum albumin in calcium lactate solution in sequence, supplementing to 800ml with water for injection, and filtering for sterilization;
s2: heating 100mL of water for injection to boil, adding polyvinylpyrrolidone, adding d-mannitol, and stirring until the mixture is dissolved and transparent; the preparation of the solution B is completed by autoclaving under the condition of 15 pounds for 15 minutes and supplementing the solution to 200mL by using water for injection;
s3: mixing 800ml of the solution A and 200ml of the solution B, and storing for 1-2 weeks at normal temperature; when in use, the protective agent and the antigen of the newcastle disease and the infectious bronchitis virus are mixed according to the ratio of 1: mixing at a ratio of 1, packaging, and lyophilizing.
Preferably, the filter sterilization described in S1 is performed with a 0.22 μm microporous filter.
Preferably, the freeze-drying in S3 includes freezing, evacuating, and drying, wherein the freezing is performed at a low temperature of-40 ℃ for 480min, the frozen heat-resistant protective agent is then evacuated at a low temperature of-40 ℃ for 1min, the evacuation environment is maintained at a vacuum degree of 90ubar, and then the drying is performed for three times in a vacuum degree of 90ubar, the temperature for the first drying is-20 ℃, the time for the first drying is 2000min, the temperature for the second drying is 20 ℃, the time for the second drying is 300min, the temperature for the third drying is 25 ℃, and the time for the third drying is 1 min; and then the fourth drying is carried out under the vacuum degree of 50ubar, the temperature of the fourth drying is 25 ℃, and the time of the fourth drying is 300-.
The application of the heat-resistant protective agent for the Newcastle disease and bronchitis combined live vaccine is as follows: when in use, the protective agent is mixed with newcastle disease and infectious bronchitis virus antigen according to the proportion of 1: 1, subpackaged and freeze-dried.
Compared with the prior art, the invention has the beneficial effects that:
after the vaccine protected by the protective agent is placed in a 37 ℃ incubator for 10 days, virus content is measured according to a potency test chick embryo test method; compared with the virus content of effectiveness test, the virus content of each feather of Newcastle disease and infectious bronchitis of chicken is reduced by not more than 1 titer after 10 days, all the test results are qualified, the performance of the protective agent is stable, and the yield of the protective agent produced by the preparation process of the heat-resistant protective agent is high.
DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all 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.
Example 1
1) Firstly, heating 500ml of water for injection to boil, slowly adding 30g of calcium lactate, stirring until the calcium lactate is completely dissolved and transparent, and filtering by qualitative filter paper; dissolving 10g of thiourea, 20g of sucrose, 4g of trehalose, 6g of L-sodium glutamate and 16g of bovine serum albumin in the calcium lactate solution in sequence, supplementing the solution to 800ml with water for injection, and filtering and sterilizing the solution to complete the preparation of the solution A;
2) heating 100mL of water for injection to boil, adding 5g of polyvinylpyrrolidone, adding 4g of d-mannitol, and stirring until the mixture is dissolved and transparent; the preparation of the solution B is completed by autoclaving under the condition of 15 pounds for 15 minutes and supplementing the solution to 200mL by using water for injection;
3) mixing the solution A, the solution B and the solution C according to the volume ratio of 4: 1 to prepare a heat-resistant protective agent;
4) mixing the vaccine freeze-drying protective agent with the newcastle disease and infectious bronchitis bigeminal live vaccine virus liquid according to the ratio of 1: 1 (V/V);
5) subpackaging and then freeze-drying;
example 2
1) Firstly, heating 500ml of water for injection to boil, slowly adding 25g of calcium lactate, stirring until the calcium lactate is completely dissolved and transparent, and filtering by qualitative filter paper; dissolving 7g of thiourea, 16g of sucrose, 5g of trehalose, 10g of L-sodium glutamate and 20g of bovine serum albumin in the calcium lactate solution in sequence, supplementing the solution to 800ml with water for injection, and filtering and sterilizing the solution to complete the preparation of the solution A;
2) heating 100mL of water for injection to boil, adding 6g of polyvinylpyrrolidone, adding 5g of d-mannitol, and stirring until the mixture is dissolved and transparent; the preparation of the solution B is completed by autoclaving under the condition of 15 pounds for 15 minutes and supplementing the solution to 200mL by using water for injection;
3) mixing the solution A, the solution B and the solution C according to the volume ratio of 4: 1 to prepare a heat-resistant protective agent;
4) mixing the vaccine freeze-drying protective agent with the newcastle disease and infectious bronchitis bigeminal live vaccine virus liquid according to the ratio of 1: 1 (V/V);
5) subpackaging and freeze-drying.
Example 3
1) Firstly, heating 500ml of water for injection to boil, slowly adding 20g of calcium lactate, stirring until the calcium lactate is completely dissolved and transparent, and filtering by qualitative filter paper; dissolving 5g of thiourea, 15g of sucrose, 5g of trehalose, 6g of L-sodium glutamate and 13g of bovine serum albumin in the calcium lactate solution in sequence, supplementing the solution to 800ml with water for injection, and filtering and sterilizing the solution to complete the preparation of the solution A;
2) heating 100mL of water for injection to boil, adding 4g of polyvinylpyrrolidone, adding 3g of d-mannitol, and stirring until the mixture is dissolved and transparent; the preparation of the solution B is completed by autoclaving under the condition of 15 pounds for 15 minutes and supplementing the solution to 200mL by using water for injection;
3) mixing the solution A, the solution B and the solution C according to the volume ratio of 4: 1 to prepare a heat-resistant protective agent;
4) mixing the vaccine freeze-drying protective agent with the newcastle disease and infectious bronchitis bigeminal live vaccine virus liquid according to the ratio of 1: 1 (V/V);
5) subpackaging and freeze-drying.
Protective agent effect test standard:
after 3 bottles of the vaccine from examples 1 to 3 were placed in an incubator at 37 ℃ for 10 days, the virus content was measured by the efficacy test chick embryo test method. Compared with the virus content of the efficacy test, the virus content of each feather chicken Newcastle disease and infectious bronchitis does not decrease by more than 1 titer, and the feather chicken Newcastle disease and infectious bronchitis virus are judged to be qualified.
Table 1 shows the results of experiments on the virus content of each feather of Newcastle disease and infectious bronchitis of chickens, which are obtained after the virus content is measured according to the effectiveness test chick embryo test method;
TABLE 1 test results
While the invention has been described above with reference to an embodiment, various modifications may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. In particular, the various features of the embodiments disclosed herein may be used in any combination, provided that there is no structural conflict, and the combinations are not exhaustively described in this specification merely for the sake of brevity and conservation of resources. Therefore, it is intended that the invention not be limited to the particular embodiments disclosed, but that the invention will include all embodiments falling within the scope of the appended claims.
Claims (6)
1. A heat-resistant protective agent for a newcastle disease and bronchitis combined live vaccine is characterized in that: consists of liquid A and liquid B according to the volume ratio of 4: 1; wherein the solution A consists of the following components: 20-30g of calcium lactate, 5-10g of thiourea, 15-20g of sucrose, 3-5g of trehalose, 5-10g of L-sodium glutamate and 10-20g of bovine serum albumin; supplementing water for injection to 800 ml; the solution B consists of the following components: 4-6g of polyvinylpyrrolidone and 3-5g of d-mannitol, and the volume is increased to 200mL by using water for injection.
2. The combined live vaccine heat-resistant protective agent for Newcastle disease and bronchitis according to claim 1, which is characterized in that: the solution A consists of the following components: 30g of calcium lactate, 10g of thiourea, 20g of sucrose, 4g of trehalose, 6g of L-sodium glutamate and 16g of bovine serum albumin; the injection water is supplemented to 800 ml.
3. The combined live vaccine heat-resistant protective agent for Newcastle disease and bronchitis according to claim 1, which is characterized in that: the solution B consists of the following components: 5g of polyvinylpyrrolidone and 4g of d-mannitol, and the volume of the solution is made up to 200mL with water for injection.
4. A preparation process of a chicken Newcastle disease and bronchitis bivalent live vaccine heat-resistant protective agent is characterized by comprising the following steps:
s1: firstly, heating 500ml of water for injection to boil, slowly adding calcium lactate, stirring until the calcium lactate is completely dissolved and transparent, and filtering by qualitative filter paper; dissolving thiourea, sucrose, trehalose, sodium L-glutamate and bovine serum albumin in calcium lactate solution in sequence, supplementing to 800ml with water for injection, filtering, and sterilizing
S2: heating 100mL of water for injection to boil, adding polyvinylpyrrolidone, adding d-mannitol, and stirring until the mixture is dissolved and transparent; the preparation of the solution B is completed by autoclaving under the condition of 15 pounds for 15 minutes and supplementing the solution to 200mL by using water for injection;
s3: mixing 800ml of the solution A and 200ml of the solution B, and storing for 1-2 weeks at normal temperature; when in use, the protective agent is mixed with newcastle disease and infectious bronchitis virus antigen according to the proportion of 1: 1, subpackaged and freeze-dried.
5. The preparation process of the Newcastle disease and bronchitis bivalent live vaccine heat-resistant protective agent according to claim 4, which is characterized in that: the filter sterilization described in S1 was performed using a 0.22 μm microporous filter.
6. The preparation process of the Newcastle disease and bronchitis bivalent live vaccine heat-resistant protective agent according to claim 4, which is characterized in that: the freeze-drying in the S3 comprises freezing, evacuating and drying, wherein the freezing is carried out for 480min in a low-temperature environment at-40 ℃, then the frozen heat-resistant protective agent is placed in a low-temperature environment at-40 ℃ and evacuated for 1min, the evacuation environment is kept at a vacuum degree of 90ubar, and then the drying is carried out for three times in a vacuum degree of 90ubar, the temperature for the first drying is-20 ℃, the time for the first drying is 2000min, the temperature for the second drying is 20 ℃, the time for the second drying is 300min, the temperature for the third drying is 25 ℃, and the time for the third drying is 1 min; and then the fourth drying is carried out under the vacuum degree of 50ubar, the temperature of the fourth drying is 25 ℃, and the time of the fourth drying is 300-.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114042167A (en) * | 2021-12-08 | 2022-02-15 | 山东滨州博莱威生物技术有限公司 | Newcastle disease and infectious bronchitis bivalent live vaccine heat-resistant protective agent and preparation method and application thereof |
| CN116983419A (en) * | 2023-08-08 | 2023-11-03 | 北京科牧丰生物制药有限公司 | Universal heat-resistant freeze-drying protective agent and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102430125A (en) * | 2011-12-31 | 2012-05-02 | 山东滨州博莱威生物技术有限公司 | Heat-resistant stabilizer for chicken embryos fluid lyophilization, as well as preparation method and application thereof |
| CN102908625A (en) * | 2011-08-01 | 2013-02-06 | 普莱柯生物工程股份有限公司 | Chicken Newcastle disease and avian infectious bronchitis bivalent live vaccine heat-resistant protective agent and preparation process thereof |
| CN104984357A (en) * | 2015-06-02 | 2015-10-21 | 长春百克生物科技股份公司 | Vaccine protective agent composition without gelatin and flu attenuated live vaccine |
-
2020
- 2020-03-23 CN CN202010206099.6A patent/CN111184871A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102908625A (en) * | 2011-08-01 | 2013-02-06 | 普莱柯生物工程股份有限公司 | Chicken Newcastle disease and avian infectious bronchitis bivalent live vaccine heat-resistant protective agent and preparation process thereof |
| CN102430125A (en) * | 2011-12-31 | 2012-05-02 | 山东滨州博莱威生物技术有限公司 | Heat-resistant stabilizer for chicken embryos fluid lyophilization, as well as preparation method and application thereof |
| CN104984357A (en) * | 2015-06-02 | 2015-10-21 | 长春百克生物科技股份公司 | Vaccine protective agent composition without gelatin and flu attenuated live vaccine |
| WO2016192487A1 (en) * | 2015-06-02 | 2016-12-08 | 长春百克生物科技股份公司 | Non-gelatin vaccine protectant composition and live attenuated influenza vaccine |
Non-Patent Citations (1)
| Title |
|---|
| 李永洲等: "兽用疫苗耐热保护剂的种类及作用机理" * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114042167A (en) * | 2021-12-08 | 2022-02-15 | 山东滨州博莱威生物技术有限公司 | Newcastle disease and infectious bronchitis bivalent live vaccine heat-resistant protective agent and preparation method and application thereof |
| CN116983419A (en) * | 2023-08-08 | 2023-11-03 | 北京科牧丰生物制药有限公司 | Universal heat-resistant freeze-drying protective agent and preparation method thereof |
| CN116983419B (en) * | 2023-08-08 | 2024-04-26 | 北京科牧丰生物制药有限公司 | Universal heat-resistant freeze-drying protective agent and preparation method thereof |
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