CN1233418C - Heat-resisting lyophilized protectant for chicken newcastle disease, chicken infectious bronchitis (H120, H52) dual lyophilized vaccine and preparing process thereof - Google Patents
Heat-resisting lyophilized protectant for chicken newcastle disease, chicken infectious bronchitis (H120, H52) dual lyophilized vaccine and preparing process thereof Download PDFInfo
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- CN1233418C CN1233418C CNB2004100295822A CN200410029582A CN1233418C CN 1233418 C CN1233418 C CN 1233418C CN B2004100295822 A CNB2004100295822 A CN B2004100295822A CN 200410029582 A CN200410029582 A CN 200410029582A CN 1233418 C CN1233418 C CN 1233418C
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Abstract
The present invention relates to a heat-resisting freeze drying protective agent of a bigeminy freeze-dried vaccine for a newcastle disease and infectious bronchitis (H120, H52) of chickens and a preparation technology thereof. The heat-resisting freeze drying protective agent of a bigeminy freeze-dried vaccine is characterized in that the preparation technology comprises: effectively components of the protective agent are respectively sterilized, and the components which can be sterilized by high pressure are dissolved in distillation water according to preparation proportion to be sterilized for 30 to 40 minutes at 116 DEG C; the components without high-temperature resistance are also dissolved in the distillation water according to the preparation proportion to be filtered and sterilized by a filter film of which the hole diameter is 0.22 mu m, and the two components are mixed to obtain the freeze drying heat-resisting protective agent. The heat-resisting freeze drying protective agent is added in antivirus raw liquid according to the proportion of 1:1 when being used and is frozen and dried after being separately loaded, products still maintain the appearance after freeze drying after being preserved for 24 months at the temperature of 2 to 8 DEG C, and the virus content exceeds the rule standard of national veterinarian biological products. The present invention provides a product having the advantages of low cost and long effect and a preparation method thereof, and the heat-resisting freeze drying protective agent solves the longtime problem of high cost of transportation and storage for the use of the veterinarian biological products.
Description
Technical field
The present invention relates to the Freeze Drying Technique of biological product, preservation adjuvant and manufacture method thereof.
Background technology
China's veterinary biological product protective agent commonly used at present is mainly 5% sucrose skim milk and 5-10% sucrose, the 1.5-3% gelatin, and goods require cryopreservation (below 15 ℃), and the storage life of 2-8 ℃ of condition only 3-9 month must use up at normal temperatures as early as possible.This is just for the preservation of veterinary biological product, transportation, use etc. bring many difficulties and inconvenience, simultaneously owing to must cryopreservation causing energy resource consumption and storage cost big.
At present, the flourishing in the world biological freeze-dried products of national veterinary almost all is that 2-8 ℃ of preservation is more than 24 months.But the not play-by-play of square method.Chinese patent CN1176827A " a kind of manufacture method of freeze-dried live vaccine for animal use " proposes to adopt with polyvinylpyrrolidone, sodium carboxymethyl cellulose; tamarind gum; sodium glutamate; trehaloses etc. are the freeze drying protectant that the formulated of main matrix freeze drying protectant becomes; the method that freeze-drying curve that employing adapts with it and newcastle disease vaccine virus antigen are made the newcastle disease freeze-dried live vaccine, but this Seedling long preservation under 2-8 ℃ of condition.
Summary of the invention
The purpose of this invention is to provide dried vaccine heat-resisting lyophilized protecting agent of a kind of infectious bronchitis of chicken H120 or H52 and newcastle disease bigeminy and preparation technology, reduce protective condition, extend using period and reduce use cost.
Infectious bronchitis of chicken H120 or H52 and newcastle disease bigeminy freeze dried vaccine is characterized in that the component proportioning of heat-resisting lyophilized protecting agent is: gelatin 2%, dextrin 6%, bovine serum albumin 1-2%, sucrose 5-6%, PVP K903%, casein peptone 1-2%, sorbitol 2%, sodium glutamate 1%, Vc0.2-0.3%; Be the dry component in the preparation, percentage ratio residue part is distilled water.
The preparation technology of the heat-resisting lyophilized protecting agent of above-mentioned vaccine presses component, mixed, it is characterized in that protectant active princlple is handled degerming respectively: to can autoclaved gelatin, dextrin, sucrose, PVPK90, casein peptone; Be dissolved in the distilled water in the preparation ratio, sterilized 30-40 minute for 116 ℃; Component bovine serum albumin, sorbitol, sodium glutamate, the Vc of non-refractory also are dissolved in the distilled water with aperture 0.22 μ m membrane filtration degerming in the preparation ratio; After finishing degerming respectively two kinds of compositions are mixed, be the lyophilizing heat resisting protective, add in the virus antigen liquid in 1: 1 ratio during use, carry out lyophilization after the packing.
The preparation of preparation technology's preparation of the present invention can be preserved more than 24 months under 2-8 ℃ of condition, can reach every index of regulation in " People's Republic of China's veterinary biological product quality standard ".
The specific embodiment
Infectious bronchitis of chicken H120 or H52 and newcastle disease bigeminy freeze dried vaccine heat-resisting lyophilized protecting agent, comprise gelatin commonly used, sucrose, it is characterized in that the weight percentages of heat-resisting lyophilized protecting agent is: gelatin 2%, dextrin 6%, bovine serum albumin 1-2%, sucrose 5-6%, PVPK90 3%, casein peptone 1-2%, sorbitol 2%, sodium glutamate 1%, Vc 0.2-0.3%; Be the dry component in the preparation, percentage ratio residue part is distilled water.
The preparation technology of infectious bronchitis of chicken H120 or H52 and newcastle disease bigeminy freeze dried vaccine heat-resisting lyophilized protecting agent, comprise dry component mixed by weight percentage with above-mentioned heat-resisting lyophilized protecting agent, it is characterized in that protectant active princlple is carried out degerming to be handled: to can autoclaved gelatin, dextrin, sucrose, PVP K90, casein peptone, be dissolved in the distilled water 116 ℃ of sterilizations 30-40 minute in ratio separately; With component sorbitol, bovine serum albumin, sodium glutamate, the Vc of non-refractory, also be dissolved in the membrane filtration degerming of using aperture 0.22 μ m in the distilled water in proportion; After finishing degerming respectively two kinds of compositions are mixed, be the lyophilizing heat resisting protective, add in the virus antigen liquid in 1: 1 ratio during use, carry out lyophilization after the packing.
Technical parameter and detection method
1. the physical behavior of product, steriling test, mycoplasma check, exogenous virus check, specificity check, safety verification, residual moisture mensuration and vacuum are measured by the method and the standard of " People's Republic of China's veterinary biological product rules " and are undertaken.
2. frozen in the cycle and be no more than 48h.
3. protectant toxicity test
Bird vaccine carries out each 20 of collunarium, eye dripping, drinking-water and subcutaneous injections by containing 10 times of amounts protecting dosage in the middle of the product respectively to 7 age in days healthy chicks, observes 20d, and chickling is all strong to live; There is not any untoward reaction.
4. viral level is measured and standard
(1) assay method:
A. vaccine is diluted to 1 plumage part/0.5mL with normal saline, is respectively charged into 2 in vitro, every pipe 1mL.The 1st pipe adds the newcastle disease antiserum of equivalent, and the 2nd pipe adds the infectious bronchitis of chicken antiserum of equivalent.At room temperature and 1h, this moment, viral level was 1 plumage part/0.1mL, i.e. 10-1.The 1st pipe continues 10 times of serial dilutions, measures the infectious bronchitis of chicken part.The 2nd pipe continues 10 times of serial dilutions, measures the newcastle disease part, and its method is with infectious bronchitis of chicken and newcastle disease freeze-dried vaccine.
B. vaccine is contained tissue mass and is diluted to 500 times by real with normal saline, be respectively charged in two test tubes, every pipe 1mL, the 1st pipe adds the newcastle disease antiserum of equivalent, and the 2nd pipe adds equivalent infectious bronchitis of chicken antiserum.Be 10-3 this moment, at room temperature in and remake 10 times of serial dilutions behind the 1h, other method is the same.
(2) reach standard
One. the standard of the weak malicious freeze-dried vaccine of newcastle disease:
Viral loss rate is less than 0.4 titre after A, the lyophilizing.
B. every plumage part viral level 〉=10 after the lyophilizing
6.7EID
50
C. the product after the lyophilizing is put 37 ℃ and is preserved 10d and 30d and carry out viral level and measure, the viral level rate of descent the former within 1 titre, the latter is within 2 titres.
D. product is put 25 ℃ of preservation 30d after the lyophilizing, and the viral level rate of descent is within 0.6 titre.
E. product is put 2-8 ℃ of preservation 24 months, every plumage part viral level 〉=10
6.17EID
50
Two. the standard of infectious bronchitis of chicken freeze-dried vaccine:
Viral loss rate is less than 0.4 titre after A, the lyophilizing.
B. H after the lyophilizing
52Every plumage part viral level 〉=10
4.5EID
50
C. product is put 37 ℃ of preservation 10d and 30d, and the viral level rate of descent is respectively within 1 to 2 titre.
D. product is put 25 ℃ of preservation 30d, and the viral level rate of descent is no more than 1 titre.
E. product is put 2-8 ℃ of preservation 24 months, H
52Every plumage part viral level 〉=10
3.5EID
50
Claims (2)
1, a kind of infectious bronchitis of chicken H120 or H52 and newcastle disease bigeminy freeze dried vaccine heat-resisting lyophilized protecting agent is characterized in that the weight percentages of heat-resisting lyophilized protecting agent is: gelatin 2%, dextrin 6%, bovine serum albumin 1-2%, sucrose 5-6%, PVP K90 3%, casein peptone 1-2%, sorbitol 2%, sodium glutamate 1%, Vc 0.2-0.3%; Be the dry component in the preparation, percentage ratio residue part is distilled water.
2, the technology of the preparation described infectious bronchitis of chicken H120 of claim 1 or H52 and newcastle disease bigeminy freeze dried vaccine heat-resisting lyophilized protecting agent, comprise dry component mixed by weight percentage with above-mentioned heat-resisting lyophilized protecting agent, it is characterized in that protectant active princlple is carried out degerming to be handled: to can autoclaved gelatin, dextrin, sucrose, PVP K90, casein peptone, be dissolved in the distilled water 116 ℃ of sterilizations 30-40 minute in ratio separately; With component sorbitol, bovine serum albumin, sodium glutamate, the Vc of non-refractory, also be dissolved in the membrane filtration degerming of using aperture 0.22 μ m in the distilled water in proportion; After finishing degerming respectively two kinds of compositions are mixed, be the lyophilizing heat resisting protective, add in the virus antigen liquid in 1: 1 ratio during use, carry out lyophilization after the packing.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100295822A CN1233418C (en) | 2001-12-21 | 2001-12-21 | Heat-resisting lyophilized protectant for chicken newcastle disease, chicken infectious bronchitis (H120, H52) dual lyophilized vaccine and preparing process thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100295822A CN1233418C (en) | 2001-12-21 | 2001-12-21 | Heat-resisting lyophilized protectant for chicken newcastle disease, chicken infectious bronchitis (H120, H52) dual lyophilized vaccine and preparing process thereof |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB011388714A Division CN1168503C (en) | 2001-12-21 | 2001-12-21 | Veterinarian virus kind biological product heat resisting freeze drying protective agent and its preparation technique |
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| Publication Number | Publication Date |
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| CN1528458A CN1528458A (en) | 2004-09-15 |
| CN1233418C true CN1233418C (en) | 2005-12-28 |
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| CNB2004100295822A Expired - Fee Related CN1233418C (en) | 2001-12-21 | 2001-12-21 | Heat-resisting lyophilized protectant for chicken newcastle disease, chicken infectious bronchitis (H120, H52) dual lyophilized vaccine and preparing process thereof |
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Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100418580C (en) * | 2005-11-14 | 2008-09-17 | 中国农业科学院哈尔滨兽医研究所 | Heat resistant freeze drying protectant for chicken Newcastle disease live vaccine and its prepn and application |
| CN101099864B (en) * | 2006-07-03 | 2010-09-29 | 河南农业大学 | Method for preparing inactivated vaccine both for preventing chicken Newcastle disease and infectious bronchitis |
| CN101514334B (en) * | 2009-03-23 | 2011-03-23 | 中国农业科学院哈尔滨兽医研究所 | Chicken infectivity bronchitis virus attenuated vaccine strain and application thereof |
| CN102908625B (en) * | 2011-08-01 | 2014-04-16 | 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 |
| CN103182082B (en) * | 2013-03-25 | 2014-08-20 | 浙江诺倍威生物技术有限公司 | Bivalent live vaccine for avian infectious bronchitis and preparation method thereof |
| CN103599533A (en) * | 2013-09-23 | 2014-02-26 | 天津瑞普生物技术股份有限公司 | Chicken new castle disease-infectious bronchitis trivalent combined vaccine and preparation method thereof |
| TWI745278B (en) | 2014-10-10 | 2021-11-11 | 以色列商艾畢克生物實驗有限公司 | Reduced foaming vaccine compositions |
| CN105233296B (en) * | 2015-11-24 | 2018-08-17 | 江苏省农业科学院 | Heat-resisting lyophilized protecting agent and its preparation method and application for duck virus hepatitis live vaccine |
| CN114042167B (en) * | 2021-12-08 | 2022-10-21 | 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 |
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