CN105999288A - Swine erysipelas live vaccine heat-resistant protective agent, preparation method and application - Google Patents
Swine erysipelas live vaccine heat-resistant protective agent, preparation method and application Download PDFInfo
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- CN105999288A CN105999288A CN201610573467.4A CN201610573467A CN105999288A CN 105999288 A CN105999288 A CN 105999288A CN 201610573467 A CN201610573467 A CN 201610573467A CN 105999288 A CN105999288 A CN 105999288A
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- vaccine
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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
- 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
- A61K39/09—Lactobacillales, e.g. aerococcus, enterococcus, lactobacillus, lactococcus, streptococcus
- A61K39/092—Streptococcus
-
- 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
- A61K47/06—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
-
- 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
-
- 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)
Abstract
The invention relates to a vaccine protective agent and a preparation method thereof, in particular to a swine erysipelas live vaccine heat-resistant protective agent and a preparation method. The heat-resistant protective agent is composed of, by weight, 3-5% of gelatin, 15-17% of lactose, 5.5-7.7% of casein and the balance of water. Compared with conventional vaccines, the heat-resistant freeze-dry protective agent for live vaccines has the advantages of high stability, long storage time, low storage conditions, small activity loss of the live vaccines, high solubility and simple raw materials and is more suitable for industrial production.
Description
Technical field
The present invention relates to a kind of live vaccine protective agent and preparation method, be specifically related to a kind of pig erysipelas live-vaccine heat-proof protective agent and system
Preparation Method.
Background technology
Pig erysipelas is one of three big infectious disease of pig, in worldwide distribution, is also a kind of zoonosis, and its cause of disease is pig erysipelas
Bacillus.China's existing pig erysipelas live vaccine, has single Seedling, Combined vaccine and trigeminy vaccine at present, and prevention swine erysipelas is played by they
Important effect.
Live vaccine is different with preservation condition according to production technology, can be divided into GPF (General Protection False agent live vaccine and heat resisting protective live vaccine.
Most domestic bacterial live vaccines product is GPF (General Protection False agent live vaccine at present, and its protective agent is mainly composed of sucrose, gelatin etc.,
Protective agent prescription is simple, preparation simplicity.Owing to antigen is temperature sensitive, if under the conditions of 2～8 DEG C, storage life only has 3～9
Individual month, so most vaccine needs to preserve below-15 DEG C.
Pig erysipelas live vaccine 2 prepared by GPF (General Protection False agent～8 DEG C of preservations, effect duration is 9 months, deposits if preserving 12 months antibacterials
Live and be only 60%；Less than-15 DEG C preservations, effect duration is 12 months.Production of vaccine, transport and use during, if
It is improper that cold chain system controls, and vaccine Conservation environment temperature raises, and frequently results in vaccine potency and declines or lost efficacy, causes immuning failure.
The loss of vaccine antigen takes place mostly in the temperature Rapid Variable Design stage in freeze-drying process, thus heat-resisting protective technology is vaccine product
The key technology that matter is stable.Finding the manufacturing inspection analysis of multi items vaccine, the vaccine of different cultivars is because of antibacterial (or antigen egg
Construction features in vain) is different, it is adaptable to its resist technology is the most different.Wherein there is the difference of heat-resisting protective agent prescription, also have system
The special requirement of standby technique are different, do not have a kind of " omnipotent " protective agent that can be common to various vaccine.If use is heat-resisting
Resist technology is incorrect, although vaccine can preserve under refrigerated conditions, but the castering action that this technology is brought to vaccine quality is just
It is difficult to embody.In reality, also the long part that runs into have employed the product of heat-resisting protective technology, but still has to strengthen using dosage
The situation of effectively immunity can be realized.
Therefore, the selection of vaccine heat-resisting protective agent prescription and the importance of Technology are the most self-evident, and it is not only the cold of vaccine
Chain transport and storage are provided convenience, and are more crucially greatly improved the effect of vaccine.
Summary of the invention
It is an object of the invention to overcome pig erysipelas live vaccine preservation condition in prior art harsh, the defect that effect duration is short, it is provided that
Planting pig erysipelas live-vaccine heat-proof protective agent and preparation method, so that its preservation condition is the gentleest, effect duration is longer, meets city
The field demand to this product.
For solving the problems referred to above, pig erysipelas live-vaccine heat-proof protective agent of the present invention be the technical scheme is that a kind of pig erysipelas is lived
Vaccine heat resisting protective, is made up of the component of following weight percentage: gelatin 2-6%, lactose 8-24%, casein
3.3-9.9%, surplus is water.
Being preferably gelatin 4%, lactose 16%, casein 6.6%, surplus is water.
Described water is water for injection.
The present invention also aims to provide a kind of protectant preparation method of pig erysipelas live-vaccine heat-proof.
The preparation method of heat-resisting lyophilized protecting agent of the present invention be the technical scheme is that a kind of pig erysipelas live-vaccine heat-proof protective agent
Preparation method, after gelatin, lactose, casein are dissolved in water, stirring until each component is thoroughly dissolved, under the conditions of 116 DEG C,
Sterilizing 30 minutes, obtains pig erysipelas live-vaccine heat-proof protective agent.
Described water is water for injection.
The present invention also aims to provide a kind of pig erysipelas live vaccine using heat resisting protective.
The pig erysipelas live-vaccine heat-proof protective agent live vaccine of the present invention be the technical scheme is that a kind of heat-resisting lyophilized protecting agent
Pig erysipelas live vaccine, aseptically, is added to antibacterial heat resisting protective in bacterial antigens, uniform according to the weight ratio of 1:1
Mixing, after shaking up, by subpackage 2.5ml suspension in each vaccine bottle, then carries out lyophilization, obtains pig erysipelas epidemic disease alive
Seedling.
Wherein said, antibacterial heat resisting protective is made up of the component of following weight percentage:: gelatin 2-6%, lactose 8-24%,
Casein 3.3-9.9%, surplus is water.
Being preferably gelatin 4%, lactose 16%, casein 6.6%, surplus is water.
Described bacterial antigens are that pig erysipelas viable bacteria (is prepared by " People's Republic of China (PRC) veterinary biological product code " two OOO versions
Obtain).
Described cryodesiccated concretely comprise the following steps: each vaccine bottle is sequentially passed through following lyophilizing program:
1, pre-freeze controls:
| Step | Design temperature DEG C | Setting time min | Persistent period min |
| 1 | 2 | 1 | 10 |
| 2 | -45 | 1 | 120 |
2, cold well is arranged:
Water vessel refrigeration design temperature sets-60 DEG C；
Forvacuum 0.14mBar, warning vacuum 0.22mBar.
3, primary drying:
| Step | Design temperature DEG C | Setting time min | Persistent period min | Control vacuum |
| 1 | -25 | 90min | 120min | 0.14mBar |
| 2 | -10 | 150min | 900min | 0.14mBar |
| 3 | 5 | 120min | 60min | 0.14mBar |
4, parsing-desiccation:
| Step | Design temperature DEG C | Setting time min | Persistent period min | Control vacuum Pa |
| 1 | 25 | 20 | 120 | 0.14mBar |
| 2 | 25 | 30 | 0 | 0mBar |
The pig erysipelas live vaccine that the live vaccine of the present invention preserves with heat resisting protective, compared with traditional vaccine, has the advantage that
1), storage temperature higher, when 2-8 DEG C, protective value is the most stable, it is simple to stores, transport and uses；2), protect
Time of depositing is long, generally can preserve more than 24 months at 2-8 DEG C, and vaccine is preserving 24
After Yue, number of viable is still above more than 90%, and conventional vaccine 2-8 DEG C preserves 12 months bacteria livings and is only 60%, this technical side
The pig erysipelas live vaccine efficacy test of case is far superior to prior art；
3), heat resisting protective decrease virus in freezing dry process various chemical factors may to goods activity
Damage, live vaccine when stored in activity change less, live vaccine is played a very good protection；
4), in heat resisting protective prescription use low molecular compound lactose more, Microbial survival rate can be improved, form homogeneous suspension,
Play moisture mitigation；Macromolecular compound casein, effect protected to microorganism, can promote that its distillation forms heat-resisting bone
Frame blocks conduction of heat and heat radiation；Gelatin hydrolysate can remove impurity protein, no antigen, without anaphylaxis, without thermal source, and
Molecular weight is little, homogenizing, soluble in water, may filter that degerming, eutectic point is-12 DEG C, and the protective effect to microorganism exceeds the brightest
Glue more than 10%；
5), protective agent formula is selected the most flexible, it is to avoid the drawback that formula components is many or complicated, and the operation letter of preparation condition gentleness
Just, it is more suitable for industrialized production.
Detailed description of the invention
Unless otherwise defined, the present invention uses all technology and implication and the common skill of the technical field of the invention of scientific terminology
The implication that art personnel are generally understood that is identical.Generally, the name that the present invention uses is well known in the art or conventional, if the most special
Indicating, material used in the embodiment of the present invention is all commercially available.In order to make technical problem solved by the invention, technical side
Case and beneficial effect are clearer, and below in conjunction with specific embodiment, the present invention is further illustrated.
Embodiment 1: the protectant preparation of pig erysipelas live-vaccine heat-proof
Accurately weigh gelatin 40g, casein 66g, lactose 160 grams, add appropriate water for injection and stir until thoroughly dissolving,
Inject water to 1L；Sterilizing 30 minutes under the conditions of 116 DEG C, cooling, obtain pig erysipelas live-vaccine heat-proof protective agent.
Embodiment 2: the protectant preparation of pig erysipelas live-vaccine heat-proof
Accurately weigh gelatin 20g, casein 33g, lactose 80 grams, add appropriate water for injection and stir until thoroughly dissolving,
Inject water to 1L；Sterilizing 30 minutes under the conditions of 116 DEG C, cooling, obtain pig erysipelas live-vaccine heat-proof protective agent.
Embodiment 3: the protectant preparation of pig erysipelas live-vaccine heat-proof
Accurately weigh gelatin 60g, casein 99g, lactose 240 grams, add appropriate water for injection and stir until thoroughly dissolving,
Inject water to 1L；Sterilizing 30 minutes under the conditions of 116 DEG C, cooling, obtain pig erysipelas live-vaccine heat-proof protective agent.
Embodiment 4: the preparation of pig erysipelas live vaccine
Aseptically, the pig erysipelas live-vaccine heat-proof protective agent prepared in embodiment 1 is added in pig erysipelas live vaccine,
Uniformly mix according to the weight ratio of 1:1, after shaking up, by subpackage 2.5ml suspension in each vaccine bottle, then carry out as follows
Lyophilization operating procedure:
1) pre-freeze controls: design temperature 2 DEG C, sets time 1min, persistent period 10min；Design temperature-45 DEG C, during setting
Between 1min, persistent period 120min；
2) cold well is arranged: water vessel refrigeration design temperature sets-60 DEG C, forvacuum 0.14mBar, warning vacuum 0.22mBar；
3) primary drying: design temperature-25 DEG C, sets time 90min, persistent period 120min, controls vacuum 0.14mBar；
Design temperature-10 DEG C, sets time 150min, persistent period 900min, controls vacuum 0.14mBar；Design temperature 5 DEG C,
Setting time 120min, persistent period 60min, control vacuum 0.14mBar；
4) parsing-desiccation: design temperature 25 DEG C, sets time 20min, persistent period 120min, controls vacuum 0.14mBar；
Design temperature 25 DEG C, sets time 30min, persistent period 0min, controls vacuum 0mBar；
Prepare pig erysipelas live vaccine.
By the pig erysipelas live-vaccine heat-proof protective agent for preparing in embodiment 2-3 according to method described in embodiment 4, prepare equally
Obtain pig erysipelas live vaccine.
Embodiment 5: product inspection
Preparing pig erysipelas live vaccine respectively according to method described in above-described embodiment 1,2,3 and embodiment 4, its lot number is successively
It is 120702,120703,120804.By three batches of pig erysipelas live vaccine, before preparing complete preservation and 2-8 DEG C of condition preserves
After 30 months to character, purely inspection, diagnostic test, count plate, safety verification, efficacy test, residual moisture mensuration,
Vacuum measures indices and detects.
Every examination criteria specifies:
1, character inspection: should be filbert Sponge Porosity agglomerate, easily departs from bottle wall, dissolves rapidly after adding diluent；
2, purely inspection: the method specified by " Chinese veterinary pharmacopoeia " is tested；
3, diagnostic test: use gelatin culture medium stab culture, should grow in wire；
4, count plate: indicate head part by label, with the Martin's agar plate containing 10% Ox blood serum do count plate (by " in
State's veterinary drug allusion quotation " method that specifies carries out), result is that every part vaccine should be not less than 7.0 × 10 containing viable bacteria8.0CFU；
5, safety verification: indicate head part by label, by vaccine 20% aluminium glue normal saline dilution, subcutaneous injection body weight 20～22g
White mice 10, every 2 parts, to observe 14, inspection mice is the most strong to live；
6, efficacy test: indicate head part by label and become 1ml to contain 1/10 using dosage, skin with 20% aluminium glue normal saline dilution
Hemostasis body weight 16～18g white mice 10, every 0.2ml (containing 1/50 using dosage).After 14 days identical together with condition
Comparison white mice 6, wherein 3 comparison white mice and immunity white mice each subcutaneous injection 1000MLD bacillus rhusiopathiae suis bacterium
Liquid, the bacillus rhusiopathiae suis bacterium solution of another 3 comparison white mice each subcutaneous injection 1MLD.Observing 10, injection 1000MLD's is right
The most dead according to white mice, the most dead 2 of the control mice of injection 1MLD, immunity white mice at least protects 8；
7, residual moisture measures: the method specified by " Chinese veterinary pharmacopoeia " is measured, and result is should be less than 4.0%；
8, vacuum measure: the method specified by " Chinese veterinary pharmacopoeia " be measured 3 batches of goods of result all should occur white or
Pink colour or purple glow.
Testing result such as following table:
Embodiment 6: pig erysipelas live vaccine storage life is tested
Preparing pig erysipelas live vaccine respectively according to method described in above-described embodiment 1,2,3 and embodiment 4, its lot number is successively
It is 120702,120703,120804.Under the conditions of three batches of pig erysipelas live vaccine are stored in 2-8 DEG C respectively 0 day, 6 months,
12 months, 18 months, 24 months, detect its count plate after 30 months.
Testing result is as follows:
Embodiment 7: pig erysipelas live vaccine storage life is tested
Preparing pig erysipelas live vaccine respectively according to method described in above-described embodiment 1,2,3 and embodiment 4, its lot number is successively
It is 120702,120703,120804.Under the conditions of three batches of pig erysipelas live vaccine are stored in 37 DEG C respectively 0 day, 7 days, 10 days,
Its count plate is detected after 15 days.
Testing result is as follows:
Embodiment 5-7 test result indicate that: the pig erysipelas live vaccine prepared by the protective agent of the present invention, can preserve being placed on 2-8 DEG C
More than 24 months.And find through the test such as safety, effect, select the freeze dried vaccine that heat-resisting protective agent prescription is made at 2-8 DEG C
Under the conditions of place 24 months, wherein respectively at the 12nd, 18,24 and 30 months detection vaccine number of viable, find vaccine exist
After preserving 24 months, number of viable is still above original more than 90%, and safety verification and efficacy test all meet quality criteria requirements.
It should be noted that, although the present invention is disclosed above with preferred embodiment, so it is not limited to the present invention, any is familiar with this
Item those skilled in the art, without departing from the spirit and scope of the present invention, any amendment, equivalent and the improvement etc. made, all should wrap
Within being contained in protection scope of the present invention.
Claims (8)
1. a pig erysipelas live-vaccine heat-proof protective agent, it is characterised in that be made up of the component of following weight percentage: gelatin 2-6%, lactose 8-24%, casein 3.3-9.9%, surplus is water.
2. protective agent as claimed in claim 1, it is characterised in that being made up of the component of following weight percentage: gelatin 4%, lactose 16%, casein 6.6%, surplus is water.
3. protective agent as claimed in claim 1 or 2, it is characterised in that described water is water for injection.
4. the protectant preparation method of pig erysipelas live-vaccine heat-proof, it is characterised in that after gelatin, lactose, casein are dissolved in water, stirring is until each component is thoroughly dissolved, under the conditions of 116 DEG C, sterilizing 30 minutes, obtain pig erysipelas live-vaccine heat-proof protective agent.
5. method as claimed in claim 4, it is characterised in that gelatin, lactose, the percentage by weight of each component of cheese egg are gelatin 2-6%, lactose 8-24%, casein 3.3-9.9%, and water is water for injection.
6. method as claimed in claim 4, it is characterised in that gelatin, lactose, the percentage by weight of each component of cheese egg are gelatin 4%, lactose 16%, casein 6.6%, and water is water for injection.
7. the pig erysipelas live vaccine using heat resisting protective, it is characterised in that aseptically, is added to antibacterial heat resisting protective in pig erysipelas live vaccine, uniformly mixes according to the weight ratio of 1:1 and shakes up, then carries out lyophilization, obtain pig erysipelas live vaccine.
8. pig erysipelas live vaccine as claimed in claim 7, it is characterised in that pre-freeze control, the setting of cold well, primary drying and four steps of parsing-desiccation are experienced in described lyophilization successively, wherein
1) pre-freeze controls: design temperature 2 DEG C, sets time 1min, persistent period 10min；Design temperature-45 DEG C, sets time 1min, persistent period 120min；
2) cold well is arranged: water vessel refrigeration design temperature sets-60 DEG C, forvacuum 0.14mBar, warning vacuum 0.22mBar；
3) primary drying: design temperature-25 DEG C, sets time 90min, persistent period 120min, controls vacuum 0.14mBar；Design temperature-10 DEG C, sets time 150min, persistent period 900min, controls vacuum 0.14mBar；Design temperature 5 DEG C, sets time 120min, persistent period 60min, controls vacuum 0.14mBar；
4) parsing-desiccation: design temperature 25 DEG C, sets time 20min, persistent period 120min, controls vacuum 0.14mBar；Design temperature 25 DEG C, sets time 30min, persistent period 0min, controls vacuum 0mBar.
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| CN201610573467.4A CN105999288A (en) | 2016-07-20 | 2016-07-20 | Swine erysipelas live vaccine heat-resistant protective agent, preparation method and application |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107929749A (en) * | 2017-11-27 | 2018-04-20 | 云南生物制药有限公司 | Pig pasteurella multocida disease live vaccine protective agent and preparation method 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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| CN102459568A (en) * | 2009-05-26 | 2012-05-16 | 先进生物营养公司 | Stable dry powder composition comprising biologically active microorganisms and/or bioactive materials and methods of making |
| CN103908665A (en) * | 2013-01-05 | 2014-07-09 | 普莱柯生物工程股份有限公司 | Vaccine composition, preparation method and application thereof |
| CN103933561A (en) * | 2014-03-07 | 2014-07-23 | 青岛易邦生物工程有限公司 | Porcine epidemic diarrhea live vaccine |
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2016
- 2016-07-20 CN CN201610573467.4A patent/CN105999288A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102459568A (en) * | 2009-05-26 | 2012-05-16 | 先进生物营养公司 | Stable dry powder composition comprising biologically active microorganisms and/or bioactive materials and methods of making |
| CN103908665A (en) * | 2013-01-05 | 2014-07-09 | 普莱柯生物工程股份有限公司 | Vaccine composition, preparation method and application thereof |
| CN103933561A (en) * | 2014-03-07 | 2014-07-23 | 青岛易邦生物工程有限公司 | Porcine epidemic diarrhea live vaccine |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107929749A (en) * | 2017-11-27 | 2018-04-20 | 云南生物制药有限公司 | Pig pasteurella multocida disease live vaccine protective agent and preparation method 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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