CN102908625A - Chicken Newcastle disease and avian infectious bronchitis bivalent live vaccine heat-resistant protective agent and preparation process thereof - Google Patents
Chicken Newcastle disease and avian infectious bronchitis bivalent live vaccine heat-resistant protective agent and preparation process thereof Download PDFInfo
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- CN102908625A CN102908625A CN2011102188242A CN201110218824A CN102908625A CN 102908625 A CN102908625 A CN 102908625A CN 2011102188242 A CN2011102188242 A CN 2011102188242A CN 201110218824 A CN201110218824 A CN 201110218824A CN 102908625 A CN102908625 A CN 102908625A
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Abstract
The invention discloses a chicken Newcastle disease and avian infectious bronchitis bivalent live vaccine heat-resistant lyophilization protective agent and a preparation process thereof. The heat-resistant lyophilization protective agent is composed of a liquid A and a liquid B according to a volume ratio of 1:1. The liquid A is composed of: 4-6g of L-sodium glutamate, 4-6g of sucrose, 1-3g of D-sorbitol, 2-4g of sodium thiosulfate, 0.1-0.3g of vitamin C, and injection water for complementing the liquid to 100mL. The liquid B is composed of: 4-6g of polyvinyl pyrrolidone, and injection water for complementing the liquid to 100mL. The protective agent provided by the invention has the advantages of simple preparation method, suitability for large-scale production, lower energy consumption, and lower cost. With the protective agent, a lyophilization time is reduced to 28h or shorter, such that application efficiency is greatly improved. The protective agent has good protection performance upon a chicken Newcastle disease and avian infectious bronchitis bivalent live vaccine. With the protective agent, the vaccine can be preserved for mother than 24 months under a temperature of 2-8 DEG C.
Description
Technical field
The present invention relates to a kind of heat resisting protective of animal virus live vaccine, refer to especially a kind of newcastle disease, infectious bronchitis of chicken bigeminal live vaccine heat resisting protective and preparation technology thereof.
Background technology
Freeze drying protectant and preparation technology thereof are key technologies during live vaccine is produced, and preservation, transportation and the use of vaccine had a significant impact.At present, in China's live vaccine, the freeze dried vaccine of living accounts for sizable ratio, and domestic protective agent commonly used is mainly milk, sucrose, gelatin etc., and its prescription is simple, and is poor to the defencive function of vaccine.Adopt the vaccine of these traditional protective agent lyophilizing; usually require goods to be kept at below-15 ℃; this causes very large inconvenience to manufacturer and user; the long preservation and the transportation that are vaccine are limited; and may cause the vaccine inactivation because the vaccine preservation is improper, render a service and reduce; do not reach the epidemic prevention purpose, cause the tremendous economic loss.
Heat-resisting lyophilized protecting agent is compared with the freeze drying protectant of routine; freeze dried vaccine has the characteristics of high temperature resistant long shelf-life; thereby the performance of heat resisting protective is more better than traditional protection agent; live vaccine can be kept under 2～8 ℃ of conditions before the deadline; even under higher room temperature, also can preserve tens days, therefore make storage, transportation and the use of live vaccine all more convenient, economical.
Developed country starts to walk early to the research of heat resisting protective, and their freeze-dried live vaccine all uses heat resisting protective basically at present, but has no play-by-play.The protectant development of domestic veterinary biologics relatively lags behind; in recent years also there are some relevant research reports in China, mainly comprises the heat-resisting lyophilized protecting agent research of newcastle disease, chicken Marek's disease, chicken bursal disease, infectious bronchitis of chicken, fowlpox and swine fever freeze-dried live vaccine etc.At present existing Chinese patent CN 1772302 A heat resistant freeze drying protectant for chicken Newcastle disease live vaccine, its preparation method and application use gelatin, lactose, arginine, sorbitol as protective agent.Chinese patent CN101612404 A discloses the heat-resisting lyophilized protecting agent of newcastle disease and infectious bronchitis of chicken bigeminal live vaccine, can preserve 24 months at 2～8 ℃.But all there is the long problem of freeze-drying time in the use procedure in these patents, affected the propagation and employment of heat resisting protective.Therefore it is simple to develop a kind of compound method, and cost is low, freeze-drying time is short, can large-scale production, the newcastle disease with good protection performance, infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent just seem particularly urgently important.
Summary of the invention
In view of this, main purpose of the present invention is to provide a kind of newcastle disease and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent, is comprised of A liquid and the B liquid volume ratio according to 1: 1; Wherein, described A liquid is comprised of following each component: Pidolidone sodium 4～6g, and sucrose 4～6g, D-glucitol 1～3g, sodium thiosulfate 2～4g, vitamin C 0.1～0.3g mends to 100mL with water for injection; Described B liquid is comprised of following each component: polyvinylpyrrolidone 4～6g, mend to 100mL with water for injection.
Preferably; in newcastle disease of the present invention and the infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent; described A liquid is comprised of following each component: the constituent of A liquid is: Pidolidone sodium 4g; sucrose 5g; D-glucitol 2g; sodium thiosulfate 2g, vitamin C 0.2g mends to 100mL with water for injection.
Preferably, in newcastle disease of the present invention and the infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent, described A liquid is comprised of following each component: Pidolidone sodium 6g; sucrose 6g, D-glucitol 3g, sodium thiosulfate 3g; vitamin C 0.3g mends to 100mL with water for injection.
In newcastle disease of the present invention and the infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent, described B liquid is comprised of following each component: polyvinylpyrrolidone 4g, mend to 100mL with water for injection.
Preferably, in newcastle disease of the present invention and the infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent, described B liquid is comprised of following each component: polyvinylpyrrolidone 6g, mend to 100mL with water for injection.
Another object of the present invention is to provide the compound method of a kind of newcastle disease and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent, may further comprise the steps:
(1) preparation of A liquid: with Pidolidone sodium, sucrose, D-glucitol, sodium thiosulfate, vitamin C fully are dissolved in rear filtration sterilization in the water for injection by described quality proportioning;
(2) preparation of B liquid: autoclaving after fully being dissolved in polyvinylpyrrolidone in the water for injection described quality proportioning;
(3) the A liquid for preparing and B liquid are mixed according to 1: 1 volume ratio after, and get final product.
Preferably, 0.22 μ m filtering with microporous membrane degerming is adopted in the aseptic filtration in the preparation method of newcastle disease of the present invention and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent, step 1).
Preferably, the autoclaving in the preparation method of newcastle disease of the present invention and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent, step 2) is sterilization 30 minutes under 116 ℃ temperature.
The present invention also provides a kind of newcastle disease and the application of infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent in preparation newcastle disease and the heat-resisting dosage form of infectious bronchitis of chicken bigeminal live vaccine.
Preferably; the application process of newcastle disease of the present invention and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent is: newcastle disease and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent and newcastle disease and infectious bronchitis of chicken bigeminal live vaccine are joined Seedling with 1: 1 volume ratio, carry out lyophilization after the packing.
More preferably, in the application process of newcastle disease of the present invention and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent, the described cryodesiccated time is less than 28 hours.
As seen from the above; the applicant adjusts the prescription of heat-resisting lyophilized protecting agent by great many of experiments; solved freeze drying protectant oversize problem consuming time in step of freeze drying; under the prerequisite of the heat-resisting protective effect that guarantees freeze drying protectant; it is simple to have obtained a kind of compound method; energy consumption is lower; cost still less can large-scale production; newcastle disease with good protection performance; infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent; particularly freeze-drying time is reduced in 28 hours, that has greatly improved heat-resisting lyophilized protecting agent is protecting newcastle disease; service efficiency in the infectious bronchitis of chicken bigeminal live vaccine.
The specific embodiment
The below is described in further detail technical scheme of the present invention with specific embodiment.It will be understood by those skilled in the art that lower without departing from the spirit and scope of the present invention and can make amendment or replace the details of technical solution of the present invention and form, but these modifications and replacing all fall in the protection domain of Ben Fanming.
Embodiment 1The preparation of newcastle disease, infectious bronchitis of chicken heat-resisting lyophilized protecting agent
Prescription 1:
The constituent of A liquid is: Pidolidone sodium 4g, and sucrose 5g, D-glucitol 2g, sodium thiosulfate 2g, vitamin C 0.2g mends to 100mL with water for injection after the dissolving fully.Through 0.22 μ m membrane filtration degerming, for subsequent use.
The constituent of B liquid is: polyvinylpyrrolidone 4g, mend to 100mL with water for injection after the dissolving fully.116 ℃ of autoclavings 30 minutes, for subsequent use.
A liquid and B liquid mix according to 1: 1 volume proportion, are the heat-resisting lyophilized protecting agent of prescription 1.
Prescription 2:
The constituent of A liquid is: Pidolidone sodium 5g, and sucrose 4g, D-glucitol 1g, sodium thiosulfate 4g, vitamin C 0.1g mends to 100mL with water for injection after the dissolving fully.Through 0.22 μ m membrane filtration degerming, for subsequent use.
The constituent of B liquid is:: polyvinylpyrrolidone 5g, mend to 100mL with water for injection after the dissolving fully.116 ℃ of autoclavings 30 minutes, for subsequent use.
A liquid and B liquid mix according to 1: 1 volume proportion, are the heat-resisting lyophilized protecting agent of prescription 2.
Prescription 3:
The constituent of A liquid is: Pidolidone sodium 6g, sucrose 6g, D-glucitol 3g, sodium thiosulfate 3g, vitamin C 0.3g, be finished CL after water for injection mend to 100mL.Through 0.22 μ m membrane filtration degerming, for subsequent use.
The constituent of B liquid is: polyvinylpyrrolidone 6g, mend to 100mL with water for injection after the dissolving fully.116 ℃ of autoclavings 30 minutes, for subsequent use.
A liquid and B liquid mix according to 1: 1 volume proportion, are the heat-resisting lyophilized protecting agent of prescription 3.
Embodiment 2The heat-resisting lyophilized protecting agent of newcastle disease, infectious bronchitis of chicken bigeminal live vaccine join Seedling and the protection potency test
1. test material and reagent
1.1 strain
Newcastle disease virus Lasota strain is available from China Veterinery Drug Inspection Office.
Avian infectious bronchitis virus H120 strain, H52 strain are available from China Veterinery Drug Inspection Office.
1.2SPF egg is available from Beijing Cimmeria laboratory animal technology company limited.
1.3 all reagent of reagent the present invention are available from Sigma company.
2. the setting of joining Seedling and lyophilization program of test group bigeminal live vaccine and freeze drying protectant
After joining Seedling with newcastle disease, infectious bronchitis of chicken live vaccine by 1: 1 volume ratio respectively according to the freeze drying protectant of three prescriptions of embodiment 1; aseptic subpackaged in cillin bottle; (1 the lyophilized preparation of namely filling a prescription is A to be respectively three groups of A, B, C; the lyophilized preparation of prescription 2 is B; the lyophilized preparation of prescription 3 is C), test respectively 01,02,03 3 batch for every group.Cartonning under-40 ℃ of conditions, goods start vacuum pump this temperature pre-freeze after 3 hours, start heater after reaching max vacuum, and the primary drying Temperature Setting is-10～-5 ℃, and the time spent is in 18 hours; The redrying Temperature Setting is 25 ℃, and the time spent is in 5 hours.Therefore, the secondary freeze-drying time only is 23 hours.If adjust temperature-time in the middle of counting in, whole lyophilization cycle also is no more than 28h.
3. test group goods technical parameter and the method for inspection
3.1 the physical behavior of product, steriling test, mycoplasma check, exogenous virus check, specificity check, safety verification, residual moisture are measured and vacuum mensuration is undertaken by method and the standard of " People's Republic of China's regulations " (2005 editions).
3.2 viral level assay method and standard: with the standard of Embryo Gallus domesticus median infective dose (EID50) as the measurement vaccine valence.The titration method is carried out according to " People's Republic of China's regulations " (2005 editions) newcastle disease, infectious bronchitis of chicken bigeminal live vaccine titer detection method.
(1) standard of the weak malicious freeze-dried vaccine of newcastle disease:
A. virus is lost less than 0.3 titre after the lyophilizing.
B. every plumage part viral level 〉=10 after the lyophilizing
7.0EID
50
C. goods are put in 37 ℃ and preserve 10d and carry out viral level and measure after the lyophilizing, and viral level descends in 1 titre.
D. goods are put in 2～8 ℃ and preserved every plumage part viral level 〉=10 24 months after the lyophilizing
6.2EID
50
(2) standard of the weak malicious freeze-dried vaccine of infectious bronchitis of chicken:
A. virus is lost less than 0.3 titre after the lyophilizing.
B. every plumage part viral level 〉=10 after the lyophilizing
4.5EID
50
C. goods are put in 37 ℃ and preserve 10d and carry out viral level and measure after the lyophilizing, and viral level descends in 1 titre.
D. goods are put in 2～8 ℃ and preserved every plumage part viral level 〉=10 24 months after the lyophilizing
3.6EID
50
4. result of the test and comparison
Table 1.37 ℃ preservation titration (EID50/ plumage part) newcastle part
Table 237 ℃ preservation titration (EID50/ plumage part) infectious bronchitis part
Table 32～8 ℃ preservation titration (EID50/ plumage part) newcastle part
Table 52～8 ℃ preservation titration (EID50/ plumage) infectious bronchitis of chicken part
Show from result of the test; under 2～8 ℃ of preservation conditions; heat-resisting lyophilized protecting agent of the present invention is suitable to the protection effect of the protection effect of newcastle disease, infectious bronchitis of chicken live vaccine and existing heat-resisting lyophilized protecting agent (patent CN101612404A); but use this protective agent greatly to shorten freeze-drying time; lyophilization cycle has improved production efficiency in 28 hours.
The result of the test explanation adopts heat-resisting lyophilized protecting agent of the present invention that newcastle, infectious bronchitis of chicken live vaccine are had good protection effect.
The above only is preferred embodiment of the present invention, and is in order to limit the present invention, within the spirit and principles in the present invention not all, any modification of doing, is equal to replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (11)
1. newcastle disease and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent is characterized in that: be comprised of A liquid and the B liquid volume ratio according to 1: 1; Wherein, described A liquid is comprised of following each component: Pidolidone sodium 4～6g, and sucrose 4～6g, D-glucitol 1～3g, sodium thiosulfate 2～4g, vitamin C 0.1～0.3g mends to 100mL with water for injection; Described B liquid is comprised of following each component: polyvinylpyrrolidone 4～6g, mend to 100mL with water for injection.
2. newcastle disease according to claim 1 and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent; it is characterized in that: described A liquid is comprised of following each component: the constituent of A liquid is: Pidolidone sodium 4g; sucrose 5g; D-glucitol 2g; sodium thiosulfate 2g; vitamin C 0.2g mends to 100mL with water for injection.
3. newcastle disease according to claim 1 and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent; it is characterized in that: described A liquid is comprised of following each component: Pidolidone sodium 6g; sucrose 6g; D-glucitol 3g; sodium thiosulfate 3g; vitamin C 0.3g mends to 100mL with water for injection.
4. newcastle disease according to claim 1 and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent, it is characterized in that: described B liquid is comprised of following each component: polyvinylpyrrolidone 4g, mend to 100mL with water for injection.
5. newcastle disease according to claim 1 and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent, it is characterized in that: described B liquid is comprised of following each component: polyvinylpyrrolidone 6g, mend to 100mL with water for injection.
6. method for preparing any one described newcastle disease of claim 1-5 and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent comprises:
(1) preparation of A liquid: with Pidolidone sodium, sucrose, D-glucitol, sodium thiosulfate, vitamin C fully are dissolved in rear filtration sterilization in the water for injection by described quality proportioning;
(2) preparation of B liquid: autoclaving after fully being dissolved in polyvinylpyrrolidone in the water for injection described quality proportioning;
(3) the A liquid for preparing and B liquid are mixed according to 1: 1 volume ratio after, and get final product.
7. method according to claim 6 is characterized in that: step 1) described in aseptic filtration adopt 0.22 μ m filtering with microporous membrane degerming.
8. method according to claim 6 is characterized in that: step 2) described in autoclaving under 116 ℃ temperature the sterilization 30 minutes.
9. each described newcastle disease of claim 1-5 and the infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent application in preparation newcastle disease and the heat-resisting dosage form of infectious bronchitis of chicken bigeminal live vaccine.
10. application according to claim 9; it is characterized in that; the described newcastle disease of claim 1-5 any one and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent and newcastle disease and infectious bronchitis of chicken bigeminal live vaccine are joined Seedling with 1: 1 volume ratio, carry out lyophilization after the packing.
11. application according to claim 10 is characterized in that, the described cryodesiccated time is less than 28 hours.
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| CN201110218824.2A CN102908625B (en) | 2011-08-01 | 2011-08-01 | Chicken Newcastle disease and avian infectious bronchitis bivalent live vaccine heat-resistant protective agent and preparation process thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104367996A (en) * | 2013-08-15 | 2015-02-25 | 普莱柯生物工程股份有限公司 | Method for producing swine pseudorabies live vaccine by using passage cell source, and product thereof |
| CN111184871A (en) * | 2020-03-23 | 2020-05-22 | 辽宁益康生物股份有限公司 | Heat-resistant protective agent for newcastle disease and bronchitis combined live vaccine and preparation process thereof |
| 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 |
| CN116440282A (en) * | 2023-05-09 | 2023-07-18 | 中海生物制药(泰州)有限公司 | Freeze-drying protective agent for live vaccine of infectious bronchitis and preparation method thereof |
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| CN1528458A (en) * | 2001-12-21 | 2004-09-15 | 卫广森 | Heat-resisting lyophilized protectant for chicken newcastle disease, chicken infectious bronchitis (H120, H52) dual lyophilized vaccine and preparing process thereof |
| US20050186221A1 (en) * | 2002-05-14 | 2005-08-25 | Bailey Reynolds | Vaccine stabilizer method |
| CN101612404A (en) * | 2009-07-29 | 2009-12-30 | 哈药集团生物疫苗有限公司 | Newcastle disease and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent |
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Patent Citations (3)
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| CN1528458A (en) * | 2001-12-21 | 2004-09-15 | 卫广森 | Heat-resisting lyophilized protectant for chicken newcastle disease, chicken infectious bronchitis (H120, H52) dual lyophilized vaccine and preparing process thereof |
| US20050186221A1 (en) * | 2002-05-14 | 2005-08-25 | Bailey Reynolds | Vaccine stabilizer method |
| CN101612404A (en) * | 2009-07-29 | 2009-12-30 | 哈药集团生物疫苗有限公司 | Newcastle disease and infectious bronchitis of chicken bigeminal live vaccine heat-resisting lyophilized protecting agent |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104367996A (en) * | 2013-08-15 | 2015-02-25 | 普莱柯生物工程股份有限公司 | Method for producing swine pseudorabies live vaccine by using passage cell source, and product thereof |
| CN104367996B (en) * | 2013-08-15 | 2017-09-05 | 普莱柯生物工程股份有限公司 | A kind of method and its product that pseudorabies disease live-vaccine is produced with subculture cell source |
| CN111184871A (en) * | 2020-03-23 | 2020-05-22 | 辽宁益康生物股份有限公司 | Heat-resistant protective agent for newcastle disease and bronchitis combined live vaccine and preparation process thereof |
| 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 |
| CN116440282A (en) * | 2023-05-09 | 2023-07-18 | 中海生物制药(泰州)有限公司 | Freeze-drying protective agent for live vaccine of infectious bronchitis and preparation method thereof |
| CN116440282B (en) * | 2023-05-09 | 2023-10-20 | 中海生物制药(泰州)有限公司 | Freeze-drying protective agent for live vaccine of infectious bronchitis and preparation method thereof |
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