CN107693497A - A kind of 50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine - Google Patents
A kind of 50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine Download PDFInfo
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- CN107693497A CN107693497A CN201711096020.3A CN201711096020A CN107693497A CN 107693497 A CN107693497 A CN 107693497A CN 201711096020 A CN201711096020 A CN 201711096020A CN 107693497 A CN107693497 A CN 107693497A
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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
- A61K39/02—Bacterial antigens
- A61K39/0241—Mollicutes, e.g. Mycoplasma, Erysipelothrix
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
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- A—HUMAN NECESSITIES
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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
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/52—Bacterial cells; Fungal cells; Protozoal cells
- A61K2039/522—Bacterial cells; Fungal cells; Protozoal cells avirulent or attenuated
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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
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
- A61K2039/552—Veterinary vaccine
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Abstract
The present invention relates to vaccine freeze-drying technical field, especially a kind of porcine mycoplasmal pneumonia live vaccine rapid freeze-drying method, including the recovery of (1) porcine mycoplasmal pneumonia live vaccine freeze-drying lactobacillus, Secondary Culture, choose product freeze-dried antigen material；(2) gelatin sucrose freeze drying protectant is prepared；(3) antigen and protective agent are sufficiently mixed uniformly；(4) the product quantitative separating that will be sufficiently mixed, is placed into freeze drying box flaggy；(5) lyophilized program is run 1. to freeze：First under conditions of 5 DEG C of refrigeration of flaggy, 40 DEG C are cooled in 1 hour, and kept for 4 hours；2. vacuum：Operation will be vacuumized in freeze drying box, made in below 0.1mbar；3. lyophilization：Vacuum is warming up to 10 DEG C under the conditions of 0.07mbar in 1 hour, and is kept for 15 hours, continues to be kept for 1 hour under the conditions of 15 DEG C afterwards；4. redrying：32 DEG C are warming up within 20 minutes, is kept for 5 hours, and continues to be warming up to 36 DEG C in 20 minutes, is kept for 2 hours；5. starting tamponade function, freeze-dried vaccine tamponade, outlet are carried out.
Description
Technical field
The present invention relates to vaccine freeze-drying technical field, specific field is a kind of porcine mycoplasmal pneumonia live vaccine rapid freeze-drying side
Method.
Background technology
In recent years, the application of porcine mycoplasmal pneumonia live vaccine is progressively paid attention to and extensive use, 10 part loading amounts of traditional mode of production are produced
The step of frequent dilution operation be present during immune use in product, hence it is evident that be unfavorable for whole nest, cluster scale inoculation it is immune
Operation.
In order to adapt to marketing development demand, 50 part loading amount products more adapt to use demand, save manpower, ensure cluster
Immunoprophylaxis uniformity.But in order to fully ensure that product Effective Antigens amount in low capacity heavy dose specification product production process,
It is necessary to establish a set of effective product freeze drying process, ensures product quality.
The content of the invention
It is an object of the invention to provide a kind of 50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine, with solution
Certainly in the prior art there is the problem of frequent dilution operation in 10 part loading amount products during immune use.
To achieve the above object, the present invention provides following technical scheme：
A kind of 50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine, comprise the following steps：
(1) recovery of porcine mycoplasmal pneumonia live vaccine freeze-drying lactobacillus, Secondary Culture, choose product freeze-dried antigen material；
(2) freeze drying protectant is prepared；
(3) antigen and protective agent are sufficiently mixed uniformly；
(4) the product quantitative separating that will be sufficiently mixed, is placed into freeze drying box flaggy；
(5) lyophilized program is run：
1. freeze：First under conditions of 5 DEG C of refrigeration of flaggy, -40 DEG C are cooled in 1 hour, and kept for 4 hours；
2. vacuum：Operation will be vacuumized in freeze drying box, made in below 0.1mbar；
3. lyophilization：Vacuum is warming up to 10 DEG C under the conditions of 0.07mbar in 1 hour, and is kept for 15 hours, rear to continue
Kept for 1 hour under the conditions of 15 DEG C；
4. redrying：32 DEG C are warming up within 20 minutes, is kept for 5 hours, and continues to be warming up to 36 DEG C in 20 minutes, is protected
Hold 2 hours；
5. starting tamponade function, freeze-dried vaccine tamponade, outlet are carried out.
50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine of the present invention, wherein, will in step (1)
Antigen count plate >=109CCU carries out product and freezed.
50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine of the present invention, wherein, freeze in step (2)
Dry protective agent is gelatin sucrose freeze drying protectant.
50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine of the present invention, wherein, the gelatin sugarcane
Sugared freeze drying protectant is prepared according to the mass percentage content of gelatin 4% and sucrose 20%.
50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine of the present invention, wherein, freeze in step (2)
Dry protective agent be completely dissolved after through 115 DEG C, autoclaving processing in 40 minutes.
50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine of the present invention, wherein, step is pressed in (3)
Antigen：Protectant volume ratio is 3：1 is sufficiently mixed uniformly.
50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine of the present invention, wherein, produced in step (4)
Product press 4ml/ bottle quantitative separatings.
Compared with prior art, the beneficial effects of the invention are as follows：
(1) present invention uses cillin bottle (7ml specifications) product loading amount as the fast of 4ml porcine mycoplasmal pneumonia live vaccine product
Quick-frozen drying method, ensure that product appearance jelly type physical behavior and rehydration are instant, living bacteria count is freezing in terms of especially lyophilized loss
It is dry forward and backward basically identical, and product rated condition preserve 18 months after still keep former living bacteria count horizontal.
(2) present invention is applied to major part part, the lyophilized production of small loading amount porcine mycoplasmal pneumonia product, major part part product adaptation city
Field immunity inoculation application, and the lyophilized run time of product is short, saves equipment energy consumption, is produced suitable for promoting.
Embodiment
The technical scheme in the embodiment of the present invention will be clearly and completely described below, it is clear that described implementation
Example only part of the embodiment of the present invention, rather than whole embodiments.It is general based on the embodiment in the present invention, this area
The every other embodiment that logical technical staff is obtained under the premise of creative work is not made, belongs to protection of the present invention
Scope.
Embodiment 1
A kind of 50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine, comprise the following steps：
(1) recovery of porcine mycoplasmal pneumonia live vaccine freeze-drying lactobacillus, Secondary Culture, choose antigen count plate >=109CCU enters
Row product freezes；
(2) gelatin sucrose freeze drying protectant is prepared, gelatin sucrose freeze drying protectant is according to gelatin 4% and the content of sucrose 20%
Prepare, through 115 DEG C after being completely dissolved, autoclaving is handled within 40 minutes；
(3) antigen is pressed：Protectant volume ratio is 3：1 is sufficiently mixed uniformly；
(4) product being sufficiently mixed is placed into freeze drying box flaggy by 4ml/ bottle quantitative separatings；
(5) lyophilized program is run：
1. freeze：First under conditions of 5 DEG C of refrigeration of flaggy, -40 DEG C are cooled in 1 hour, and kept for 4 hours；
2. vacuum：Operation will be vacuumized in freeze drying box, made in below 0.1mbar；
3. lyophilization：Vacuum is warming up to 10 DEG C under the conditions of 0.07mbar in 1 hour, and is kept for 15 hours, rear to continue
Kept for 1 hour under the conditions of 15 DEG C；
4. redrying：32 DEG C are warming up within 20 minutes, is kept for 5 hours, and continues to be warming up to 36 DEG C in 20 minutes, is protected
Hold 2 hours；
5. starting tamponade function, freeze-dried vaccine tamponade, outlet are carried out.
Embodiment 2
Using the method for embodiment 1 to (168 plants) the progress rapid freeze-drying experiments of Nanjing day nation porcine mycoplasmal pneumonia live vaccine
Comprise the following steps that：
(1) product antigen is using the recovery of Nanjing day nation porcine mycoplasmal pneumonia live vaccine (168 plants) freeze-drying lactobacillus, passage training
Support, antigen count plate >=109CCU carries out product and freezed.
(2) freeze drying protectant is prepared：By gelatin 4%, the mass percentage content of sucrose 20% is prepared, and is passed through after being completely dissolved
115 DEG C of 40 minutes autoclavings processing.
(3) product matches：Antigen and protective agent are pressed 3：1 volume ratio is sufficiently mixed uniformly.
(4) packing, inlet：By the product being sufficiently mixed by 4ml/ bottle quantitative separatings, put put freeze drying box flaggy in time.
(5) operation is freezed：Packing runs lyophilized program after terminating.
1. freeze：First under conditions of 5 DEG C of refrigeration of flaggy, -40 DEG C are cooled in 1 hour, and kept for 4 hours；
2. vacuum：Operation will be vacuumized in freeze drying box, made in below 0.1mbar；
3. lyophilization：Vacuum is warming up to 10 DEG C under the conditions of 0.07mbar in 1 hour, and is kept for 15 hours, rear to continue
Kept for 1 hour under the conditions of 15 DEG C；
4. redrying：32 DEG C are warming up within 20 minutes, is kept for 5 hours, and continues to be warming up to 36 DEG C in 20 minutes, is protected
Hold 2 hours；
5. starting tamponade function, freeze-dried vaccine tamponade, outlet are carried out.
Final effect evaluation is as shown in table 1：
Table 1
For prominent beneficial effects of the present invention, following comparative example experiment has also been carried out.
Comparative example 1
Rapid freeze-drying experiment is carried out to Nanjing day nation porcine mycoplasmal pneumonia live vaccine using the method for embodiment 1, wherein, will
Lyophilization and redrying parameter adjustment in step (5) are：
Lyophilization：Vacuum is warming up to 8 DEG C under the conditions of 0.07mbar in 1 hour, and is kept for 13 hours, rear to continue
Kept for 1 hour under the conditions of 10 DEG C；
Redrying：13 DEG C are warming up within 20 minutes, is kept for 5 hours, and continues to be warming up to 36 DEG C in 20 minutes, keeps 2
Hour.
Remaining parameter keeps constant.
Test result indicates that：
| Embodiment 2 | Comparative example 1 |
| Outward appearance jelly type is good | Remaining moisture content is higher, unqualified |
Comparative example 2
Rapid freeze-drying experiment is carried out to Nanjing day nation porcine mycoplasmal pneumonia live vaccine using the method for embodiment 1, wherein, will
The mass percentage content of gelatin is adjusted to 3% in step (2), and remaining parameter keeps constant.
Comparative example 3
Rapid freeze-drying experiment is carried out to Nanjing day nation porcine mycoplasmal pneumonia live vaccine using the method for embodiment 1, wherein, will
The mass percentage content of gelatin is adjusted to 5% in step (2), and remaining parameter keeps constant.
Comparative example 4
Rapid freeze-drying experiment is carried out to Nanjing day nation porcine mycoplasmal pneumonia live vaccine using the method for embodiment 1, wherein, will
The mass percentage content of gelatin is adjusted to 6% in step (2), and remaining parameter keeps constant.
Test result indicates that：
Although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of changes, modification can be carried out to these embodiments, replace without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (7)
- A kind of 1. 50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine, it is characterised in that comprise the following steps：(1) recovery of porcine mycoplasmal pneumonia live vaccine freeze-drying lactobacillus, Secondary Culture, choose product freeze-dried antigen material；(2) freeze drying protectant is prepared；(3) antigen and protective agent are sufficiently mixed uniformly；(4) the product quantitative separating that will be sufficiently mixed, is placed into freeze drying box flaggy；(5) lyophilized program is run：1. freeze：First under conditions of 5 DEG C of refrigeration of flaggy, -40 DEG C are cooled in 1 hour, and kept for 4 hours；2. vacuum：Operation will be vacuumized in freeze drying box, made in below 0.1mbar；3. lyophilization：Vacuum is warming up to 10 DEG C under the conditions of 0.07mbar in 1 hour, and is kept for 15 hours, continues afterwards 15 Kept for 1 hour under the conditions of DEG C；4. redrying：32 DEG C are warming up within 20 minutes, is kept for 5 hours, and continues to be warming up to 36 DEG C in 20 minutes, holding 2 is small When；5. starting tamponade function, freeze-dried vaccine tamponade, outlet are carried out.
- 2. 50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine according to claim 1, it is characterised in that： Wherein, by antigen count plate >=10 in step (1)9CCU carries out product and freezed.
- 3. 50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine according to claim 2, it is characterised in that： Wherein, freeze drying protectant is gelatin sucrose freeze drying protectant in step (2).
- 4. 50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine according to claim 3, it is characterised in that： The gelatin sucrose freeze drying protectant is prepared according to the mass percentage content of gelatin 4% and sucrose 20%.
- 5. 50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine according to claim 4, it is characterised in that： Wherein, through 115 DEG C after freeze drying protectant is completely dissolved in step (2), autoclaving is handled within 40 minutes.
- 6. 50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine according to claim 5, it is characterised in that： Wherein, step (3) presses antigen：Protectant volume ratio is 3：1 is sufficiently mixed uniformly.
- 7. 50 part product rapid freeze-drying methods of porcine mycoplasmal pneumonia live vaccine according to claim 6, it is characterised in that： Wherein, product presses 4ml/ bottle quantitative separatings in step (4).
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711096020.3A CN107693497B (en) | 2017-11-03 | 2017-11-03 | Quick freeze-drying method for 50-part live vaccine of mycoplasma hyopneumoniae |
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|---|---|---|---|
| CN201711096020.3A CN107693497B (en) | 2017-11-03 | 2017-11-03 | Quick freeze-drying method for 50-part live vaccine of mycoplasma hyopneumoniae |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109078003A (en) * | 2018-08-02 | 2018-12-25 | 南京大爻网络科技有限公司 | A kind of big specification production method of mycoplasma hyopneumoniae live vaccine |
| CN110801436A (en) * | 2019-11-19 | 2020-02-18 | 畜科生物工程有限公司 | Porcine transmissible gastroenteritis and porcine epidemic diarrhea bivalent live vaccine freeze-drying protective agent and bivalent live vaccine |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102732473A (en) * | 2011-04-08 | 2012-10-17 | 华中农业大学 | Recombinant salmonella choleraesuis expressing mycoplasma hyopneumoniae p46 protein, preparation method and application thereof |
-
2017
- 2017-11-03 CN CN201711096020.3A patent/CN107693497B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102732473A (en) * | 2011-04-08 | 2012-10-17 | 华中农业大学 | Recombinant salmonella choleraesuis expressing mycoplasma hyopneumoniae p46 protein, preparation method and application thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109078003A (en) * | 2018-08-02 | 2018-12-25 | 南京大爻网络科技有限公司 | A kind of big specification production method of mycoplasma hyopneumoniae live vaccine |
| CN109078003B (en) * | 2018-08-02 | 2021-02-26 | 南京大爻网络科技有限公司 | Large-scale production method of mycoplasma hyopneumoniae live vaccine |
| CN110801436A (en) * | 2019-11-19 | 2020-02-18 | 畜科生物工程有限公司 | Porcine transmissible gastroenteritis and porcine epidemic diarrhea bivalent live vaccine freeze-drying protective agent and bivalent live vaccine |
| CN110801436B (en) * | 2019-11-19 | 2021-06-22 | 畜科生物工程有限公司 | Porcine transmissible gastroenteritis and porcine epidemic diarrhea bivalent live vaccine freeze-drying protective agent and bivalent live vaccine |
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