CN106668867A - Mumps vaccine freeze-drying protective additive free of gelatin and human albumin - Google Patents
Mumps vaccine freeze-drying protective additive free of gelatin and human albumin Download PDFInfo
- Publication number
- CN106668867A CN106668867A CN201611267267.2A CN201611267267A CN106668867A CN 106668867 A CN106668867 A CN 106668867A CN 201611267267 A CN201611267267 A CN 201611267267A CN 106668867 A CN106668867 A CN 106668867A
- Authority
- CN
- China
- Prior art keywords
- vaccine
- finished product
- mumps
- semi
- freeze
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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
- A61K39/12—Viral antigens
-
- 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
- A61K47/02—Inorganic compounds
-
- 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
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/12—Carboxylic acids; Salts or anhydrides thereof
-
- 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
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
- A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
-
- 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
- A61K47/22—Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
-
- 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
- A61K2039/5254—Virus avirulent or attenuated
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
- C12N2760/00011—Details
- C12N2760/18011—Paramyxoviridae
- C12N2760/18111—Avulavirus, e.g. Newcastle disease virus
- C12N2760/18134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Epidemiology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- General Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Virology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Inorganic Chemistry (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention relates to the field of biological products, in particular to a mumps vaccine freeze-drying protective additive free of gelatin and human albumin. The umps vaccine freeze-drying protective additive free of the gelatin and human albumin comprises sucrose, sodium glutamate, urea, disodium hydrogen phosphate and citric acid and is characterized in that the final concentration of the sucrose in a mumps vaccine semi-finished product is 30-100g/L, the final concentration of the sodium glutamate in the mumps vaccine semi-finished product is 0.5-15g/L, the final concentration of the urea in the mumps vaccine semi-finished product is 0-10g/L, the final concentration of the disodium hydrogen phosphate in the mumps vaccine semi-finished product is 1-20g/L, the final concentration of the citric acid in the mumps vaccine semi-finished product is 0.2-10g/L, matrix liquid for preparing the protective agent is a 199 culture solution, a PBS buffer solution or injection water, and the vaccine semi-finished product is a liquid vaccine before freeze drying. The invention further provides application of the freeze-drying protective additive. When the freeze-drying protective additive is used for preparing the freeze-dried vaccine, the stability of the vaccine during freeze drying and storage can be increased, the safety of the freeze-dried vaccine to human bodies is increased greatly, and the untoward effect of the vaccine is reduced.
Description
Technical field
The present invention relates to field of biological product, and in particular to a kind of mumps Vaccine without gelatin and Human Albumin is protected
The formula and using method of shield agent.
Background technology
The application of preventative vaccine makes common epidemic infectious diseases obtain effective control, effectively reduces these infections
The M ＆ M of disease.But, find that most of vaccines have higher untoward reaction during the use of vaccine
Incidence rate；Extensive application with vaccine and the further investigation to untoward reaction inducement find occur after many vaccinations
Untoward reaction problem has the freeze drying protectant in direct relation, particularly freeze dried vaccine product with the protective agent of vaccine product, its
In gelatin or gelatine derivative can directly cause the allergy such as vaccinee's allergy, research shows the mistake of human body caused by gelatin
Quick reaction has humoral immune mechanisms (IgE antibody), and (Zhang Zhuoguang, the sensitization of cattle source gelatin is made in vaccine also cellular immune mechanism
With,《Foreign medical science》, (the 2nd phase) in 2002,25).
It is standard production that existing vaccine product majority is the biological product code issued with reference to World Health Organization (WHO), is marked
Protectant gelatin usage amount is not any limitation as in standard.Japanese FDA in 1986 approval vaccine listing license and
In the vaccine listing license of U.S. FDA approval, not using gelatin as the disabling composition for producing vaccine.In the patent text of early stage
In offering, most of vaccine protectants all contain gelatin.
Chinese Pharmacopoeia Commission is distinctly claimed " as stable on July 17th, 2007 to July 19 5 Beijing Meeting summary
The gelatin of agent composition is expelled to after human body can occur anaphylaxiss, and manufacturing enterprise should carry out replacing the correlational study work of gelatin
Make, further to improve the safety of product ".
At present, human serum albumin is widely used Virus culture and freeze drying protectant in vaccines arts, human seralbumin egg
It is in vain to extract from the blood plasma of people, such mode of production is not only limited by blood plasma supply, but also may be contained dangerous
Infectious disease pathogens, such as hepatitis viruss etc., therefore there is larger worry during use；Additionally, human serum albumin
It is expensive so that the cost of vaccine rises sharply.So forcing a series of succedaneum that researcher studies human serum albumins
Improve traditional formula.
The zest to human body is although reduced without gelatin, the protective agent of human serum albumin in currently available technology,
But it is unsatisfactory to the protected effect of vaccine so that stability of the vaccine in freeze-drying process and during storage declines, easily
Fail.Therefore, a kind of safely and effectively gelatin-free is badly in need of in this area, without human serum albumin's vaccine freeze-drying protective agent,
While vaccine quality is ensured, immune risk and production cost are reduced.
The content of the invention
It is an object of the invention to provide it is a kind of without gelatin, the new generation vaccine freeze drying protectant without Human Albumin, make
Endotoxin content declines, and is not susceptible to anaphylaxiss, and still can keep higher stability and longer effect duration.
In order to achieve the above object, the present invention provides a kind of vaccine freeze-drying protective agent, and each composition of the freeze drying protectant is in epidemic disease
It is final concentration of in Seedling semi-finished product：Sucrose 30-100g/L, sodium glutamate 0.5-15g/L, carbamide 0-10g/L, disodium hydrogen phosphate 1-
20g/L, citric acid 0.2-10g/L；The vaccine semi-finished product refer to the liquid vaccine before lyophilizing.
Preferably, each composition of the freeze drying protectant is final concentration of in vaccine semi-finished product：Sucrose 30-80g/L, glutamic acid
Sodium 0.5-10g/L, carbamide 0-5g/L, disodium hydrogen phosphate 1-10g/L, citric acid 0.2-5g/L.
It is highly preferred that each composition of the freeze drying protectant is final concentration of in vaccine semi-finished product：Sucrose 50g/L, glutamic acid
Sodium 10g/L, carbamide 5g/L, disodium hydrogen phosphate 10g/L, citric acid 5g/L.
The mumps Vaccine freeze drying protectant that the present invention is provided, without gelatin and Human Albumin in its composition.
Matrix liquid for preparing freeze drying protectant of the present invention is 199 culture fluid or PBS buffer solution or water for injection.
Further, 199 described culture fluid are without phenol red 199 culture medium.
The invention provides application of the above-mentioned mumps Vaccine freeze drying protectant in lyophilizing mumps Vaccine is prepared.
Further, the invention provides a kind of method for preparing lyophilizing mumps Vaccine, comprises the following steps：
Mumps viruss liquid is provided；
Mumps Vaccine freeze drying protectant of the present invention is proportionally added into into virus liquid, mumps viruss work is made
Vaccinogen liquid；
By live mumps virus vaccine stock solution press in vaccine semi-finished product target viral titre add appropriate diluent preparing into
Vaccine semi-finished product, make each composition in vaccine freeze-drying protective agent final concentration of in vaccine semi-finished product：Sucrose 30-100g/L, paddy
Propylhomoserin sodium 0.5-15g/L, carbamide 0-10g/L, disodium hydrogen phosphate 1-20g/L, citric acid 0.2-10g/L；The diluent contains
With the freeze drying protectant identical composition of the present invention, and the concentration of each composition and freeze drying protectant of the present invention are in vaccine semi-finished product
Final concentration it is consistent；The vaccine semi-finished product are the liquid vaccine before lyophilizing；The target viral titre is 5.8 ± 0.3lg
CCID50/ml；
Vaccine semi-finished product under the conditions of 2-8 DEG C, subpackage, lyophilizing.
The condition of lyophilizing is：- 45 DEG C~-48 DEG C of pre-freeze stage minimum temperature, reaches and maintained 2 hours after minimum temperature；Rise
Magnificent -35 DEG C of drying stage final temperature, the time for reaching final temperature is 17 hours, and vacuum pressure is controlled in 4-6Pa；Solution is blotted
28 DEG C of dry stage final temperature, vacuum is not controlled, and is run 8 hours.
A kind of lyophilizing mumps Vaccine that the above-mentioned preparation method provided using the present invention is prepared falls within the present invention
Protection domain.
The invention provides a kind of freeze drying protectant of gelatin-free without Human Albumin, it being capable of rationally replacing gelatin and people
Blood albumin, and like product identical effectiveness can be reached, and with higher safety.Freeze drying protectant in the present invention
Safety and the less susceptible generation anaphylaxiss of commercialized product, safety is higher, and the quality of vaccine is far above《Chinese Pharmacopoeia》
Three required standards, its effect includes：
(1) gelatin, Human Albumin, dextran, trehalose are not contained in protective agent composition, is difficult to cause allergy anti-
Answer, such as urticaria, skin pruritus, heating, nausea, the safety of vaccine is higher, reduces vaccinated untoward reaction；
(2) the attenuated mumps vaccine, live finished product virus titer prepared using the present invention is all higher than 4.8lgCCID50/ ml, matter
Amount standard is higher than《Chinese Pharmacopoeia》Three requirements are not less than 4.0lgCCID50/ ml standards；Heat stabilization test result is all higher than
4.4lgCCID50/ ml, quality standard is higher than《Chinese Pharmacopoeia》Three requirements are not less than 4.0lgCCID50/ ml standards；Virus titer
Drop-out value is not higher than 0.6lg, and quality standard is higher than《Chinese Pharmacopoeia》Three require to decline not higher than 1.0lg standards；
(3) moisture is below 2.0%, and quality standard is higher than《Chinese Pharmacopoeia》Three less demanding in 3.0% standard；
(4) the attenuated mumps vaccine, live finished product accelerated stability and long-time stability result for being prepared using the present invention be not low
In the control sample containing gelatin and Human Albumin；
(5) endotoxin content of the attenuated mumps vaccine, live finished product prepared using the present invention is well below 2015 editions《In
State's pharmacopeia》Three require (should be not higher than 50EU/ agent)；
(6) attenuated mumps vaccine, live finished product carries out Cavia porcelluss whole body active hypersensitive test：Test sample subcutaneous injection sensitization
Afterwards again intravenous administration is excited, and observes systemic anaphylaxis produced after Cavia porcelluss injection test sample, and anaphylaxiss should be negative.
Specific embodiment
Following examples further illustrate present disclosure, but should not be construed as limiting the invention.Without departing substantially from
In the case of spirit of the invention and essence, the modification made to the inventive method, step or condition or replacement belong to the present invention
Scope.
If not specializing, the conventional meanses that technological means used are well known to those skilled in the art in embodiment；
If not specializing, agents useful for same is commercially available in embodiment.Mumps viruss S79Strain derives from Ministry of Public Health Shanghai biological product
Institute.
Mumps viruss S in the embodiment of the present invention of embodiment 179The preparation method of strain virus liquid
1. the hatching SPF hatching egg of 9~11 days is dissected, discarded Embryo Gallus domesticus head and internal organs, prepared by piece of tissue digestion thin
Born of the same parents' suspension, suitable cell concentration is diluted to cell nutrient solution by cell suspension, is sub-packed in 10L rolling bottles and is placed in 36.5 ± 1 DEG C
Thermostatic chamber culture, cultivates to growing up to fine and close cell monolayer.
2. after cell grows up to fine and close monolayer, abandon cell nutrient solution and change the viral growth liquid of pH value 7.2~7.4, press
MOI0.001~0.01 is inoculated with mumps viruss (S79Strain), it is placed in 33 ± 1 DEG C of thermostatic chambers and continues to cultivate.
3. Virus culture discards viral growth liquid after 44~52 hours, and with 0.85% brine cell surface
Bovine serum albumin is removed, is added after washing and is not contained bovine serum albumin, the viral maintaining liquid of pH value 7.4~7.6, it is placed in 33 ±
Continue to cultivate in 1 DEG C.
4. Microscopic observation harvests virus liquid when pathological changes reach more than 75%, and single harvest liquid is after merging.
The component of the freeze drying protectant of the present invention of embodiment 2, content (final concentrations in vaccine semi-finished product)
Component 1：Sucrose, pharmaceutical grade, 30~100g/L；
Component 2：Sodium glutamate, pharmaceutical grade, 0.5~15g/L；
Component 3：Carbamide, pharmaceutical grade, 0.0~5.0g/L；
Component 4：Disodium hydrogen phosphate, analyzes pure, 1.0~20g/L；
Component 5：Citric acid, pharmaceutical grade, 0.2~10g/L；
Solvent：Water for injection or PBS buffer solution or 199 culture fluid；
The compound method of freeze drying protectant of the present invention：
(1) after said components 1 are weighed, solvent dissolving is added, determines molten laggard horizontal high voltage steam sterilization, sterilising conditions are
121 DEG C, 30 minutes；
(2) said components 2, component 3 are weighed, adds solvent dissolving, fixed molten rear liquid to pass through the degerming mistake of 0.2 micron membrane filter
Filter, is placed in 2~8 DEG C of freezers, standby；
(3) said components 4, component 5 are weighed, solvent dissolving is separately added into, then mixes fixed molten, fixed molten rear liquid passes through
0.2 micron membrane filter aseptic filtration, is placed in 2~8 DEG C of freezers, standby；
(4) protective agent is being mixed (1) (2) (3) using front.
Embodiment 3
With mumps viruss S79Work seed inoculation primary chick embryo cell prepared by strain, Jing after culture, harvesting virus liquid, will
Freeze drying protectant presses 1 with virus harvest liquid:The ratio of 4 (V/V) is mixed with vaccinogen liquid, wherein, freeze drying protectant is in vaccine
Final concentration of sucrose 50g/L in stock solution, sodium glutamate 10g/L, carbamide 5g/L, disodium hydrogen phosphate 10g/L, citric acid 5g/L.
The diluent of appropriate pH6.8~7.0 is added in vaccinogen liquid so that target viral titre is 5.8lgCCID50/
Ml, that is, obtain semi-finished product solution；Wherein, diluent is containing sucrose 50g/L, sodium glutamate 10g/L, carbamide 5g/L, phosphoric acid hydrogen
The solution of disodium 10g/L, citric acid 5g/L.
Semi-finished product are sub-packed in into lyophilizing in cillin bottle, as attenuated mumps vaccine, live finished product.
Freeze drying process is：- 45 DEG C of pre-freeze phase temperature, reaches and maintained 2 hours after temperature；Sublimation drying stage most final temperature
- 35 DEG C of degree, the time for reaching final temperature is 17 hours, and vacuum pressure is controlled in 5 ± 1Pa；Adsorption stripping and dry stage final temperature
28 DEG C, vacuum is not controlled, and is run 8 hours.
Embodiment 4
With mumps viruss S79Work seed inoculation primary chick embryo cell prepared by strain, Jing after culture, harvesting virus liquid, will
Freeze drying protectant presses 1 with virus harvest liquid:The ratio of 4 (V/V) is mixed with vaccinogen liquid, wherein, freeze drying protectant is in vaccine
Final concentration of sucrose 30g/L in stock solution, sodium glutamate 3g/L, carbamide 2g/L, disodium hydrogen phosphate 5g/L, citric acid 2g/L.
The diluent of appropriate pH6.8~7.0 is added in vaccinogen liquid so that target viral titre is 5.8lg CCID50/
Ml, that is, obtain semi-finished product solution；Wherein, diluent is containing sucrose 30g/L, sodium glutamate 3g/L, carbamide 2g/L, phosphoric acid hydrogen two
The solution of sodium 5g/L, citric acid 2g/L.
Semi-finished product are sub-packed in into lyophilizing in cillin bottle, as attenuated mumps vaccine, live finished product.
Freeze drying process is：- 45 DEG C of pre-freeze phase temperature, reaches and maintained 2 hours after temperature；Sublimation drying stage most final temperature
- 35 DEG C of degree, the time for reaching final temperature is 17 hours, and vacuum pressure is controlled in 5 ± 1Pa；Adsorption stripping and dry stage final temperature
28 DEG C, vacuum is not controlled, and is run 8 hours.
Embodiment 5
With mumps viruss S79Work seed inoculation primary chick embryo cell prepared by strain, Jing after culture, harvesting virus liquid, will
Freeze drying protectant presses 1 with virus harvest liquid:The ratio of 4 (V/V) is mixed with vaccinogen liquid, wherein, freeze drying protectant is in vaccine
Final concentration of sucrose 80g/L in stock solution, sodium glutamate 2g/L, disodium hydrogen phosphate 3g/L, citric acid 0.2g/L.
The diluent of appropriate pH6.8~7.0 is added in vaccinogen liquid so that target viral titre is 5.8lgCCID50/
Ml, that is, obtain semi-finished product solution；Wherein, diluent is containing sucrose 80g/L, sodium glutamate 2g/L, disodium hydrogen phosphate 3g/L, Chinese holly
The solution of rafter acid 0.2g/L.
Semi-finished product are sub-packed in cillin bottle, lyophilizing, as attenuated mumps vaccine, live finished product.
Freeze drying process is：- 45 DEG C of pre-freeze phase temperature, reaches and maintained 2 hours after temperature；Sublimation drying stage most final temperature
- 35 DEG C of degree, the time for reaching final temperature is 17 hours, and vacuum pressure is controlled in 5 ± 1Pa；Adsorption stripping and dry stage final temperature
28 DEG C, vacuum is not controlled, and is run 8 hours.
Attenuated mumps vaccine, live finished product obtained in 3~5 embodiments places different times respectively at 37 DEG C, 4 DEG C to more than,
Sampling carries out outward appearance, moisture and titration of virus, and the commercialized product of (Dalian) Vaccine Technology Co., Ltd emerging with section (contains gelatin
And human serum albumin) attenuated mumps vaccine, live product contrasted.
Quality standard is as follows：
(1) outward appearance：Faint yellow loosening body, is weak yellow liquid after redissolution, foreign.
(2) moisture：According to《Pharmacopoeia of People's Republic of China》Three (D of annex Ⅻ) is checked, should be not higher than 3.0%.
(3) titration of virus：Titration of virus detection is carried out using Microdose cytopathic effect assay.3 bottles of mixing of vaccine are taken, it is appropriate to carry out
Dilution, per dilution factor virus liquid Vero cells are connect, and put 36.5 ± 1 DEG C, 5%CO2Culture, cultivates 8 days result of determination, virus titer
4.0lg CCID should be not less than50/ml.The results are shown in Table 1, table 2.
Table 1 attenuated mumps vaccine, live, 37 DEG C of accelerated stability results
※For the attenuated mumps vaccine, live batch number that emerging (Dalian) Vaccine Technology Co., Ltd of section has listed.
Table 2 attenuated mumps vaccine, live, 2~8 DEG C of long-time stability results
※For the attenuated mumps vaccine, live batch number that emerging (Dalian) Vaccine Technology Co., Ltd of section has listed.
The finished product of the protective agent production of 3~5 embodiments is respectively at carrying out safety evaluatio, and (Dalian) emerging with section to more than
The attenuated mumps vaccine, live product of the commercialized product (containing gelatin and human serum albumin) of Vaccine Technology Co., Ltd is carried out
Contrast.
Evaluation criterion is as follows：
(1) baterial endotoxin test：According to《Pharmacopoeia of People's Republic of China》Three (the E gel limiting tests of annex Ⅻ) is examined
Look into, 50EU/ agent should be not higher than.
(2) Cavia porcelluss whole body active hypersensitive test：Again intravenous administration is excited after test sample subcutaneous injection sensitization, observes globefish
Produced systemic anaphylaxis after Mus injection test sample, anaphylaxiss should be negative.Result of the test such as table 3 below：
The attenuated mumps vaccine, live safety evaluatio of table 3
| Embodiment | Baterial endotoxin test | Cavia porcelluss whole body active hypersensitive test |
| 3 | <1.25 | It is negative |
| 4 | <1.25 | It is negative |
| 5 | <1.25 | It is negative |
| 201210003※ | <12.5 | It is negative |
※For the attenuated mumps vaccine, live batch number that emerging (Dalian) Vaccine Technology Co., Ltd of section has listed
By data above as can be seen that the present invention's is conventional use of with current without gelatin and Human Albumin protective agent
Compare with Human Albumin protective agent containing gelatin, there was no significant difference for stability result, but the vaccine freeze-drying of the present invention is protected
Agent improves the safety of vaccine, it is possible to decrease the untoward reaction of vaccine.
Although above having used general explanation, specific embodiment and test, the present invention is made to retouch in detail
State, but on the basis of the present invention, it can be made some modifications or improvements, this is to those skilled in the art apparent
's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed
Scope.
Claims (10)
1. a kind of mumps Vaccine freeze drying protectant is characterized in that, end of each composition of the freeze drying protectant in vaccine semi-finished product
Concentration is：Sucrose 30-100g/L, sodium glutamate 0.5-15g/L, carbamide 0-10g/L, disodium hydrogen phosphate 1-20g/L, citric acid
0.2-10g/L；The vaccine semi-finished product refer to the liquid vaccine before lyophilizing.
2. mumps Vaccine freeze drying protectant as claimed in claim 1, it is characterised in that each composition of the freeze drying protectant is in epidemic disease
It is final concentration of in Seedling semi-finished product：Sucrose 30-80g/L, sodium glutamate 0.5-10g/L, carbamide 0-5g/L, disodium hydrogen phosphate 1-
10g/L, citric acid 0.2-5g/L.
3. mumps Vaccine freeze drying protectant as claimed in claim 1, it is characterised in that each composition of the freeze drying protectant is in epidemic disease
It is final concentration of in Seedling semi-finished product：Sucrose 50g/L, sodium glutamate 10g/L, carbamide 5g/L, disodium hydrogen phosphate 10g/L, citric acid
5g/L。
4. the mumps Vaccine freeze drying protectant as described in claim 1-3 is arbitrary, it is characterised in that white without gelatin and human blood
Albumen.
5. the mumps Vaccine freeze drying protectant as described in claim 1-3 is arbitrary, it is characterised in that prepare protectant substrate
Liquid is 199 culture fluid or PBS buffer solution or water for injection.
6. mumps Vaccine freeze drying protectant as claimed in claim 1, it is characterised in that 199 described culture fluid are without phenol
Red 199 culture medium.
7. the mumps Vaccine freeze drying protectant described in any one of claim 1-6 prepare lyophilizing mumps Vaccine in should
With.
8. application as claimed in claim 7, it is characterised in that：Comprise the following steps：
Mumps viruss liquid is provided；
The mumps Vaccine freeze drying protectant described in any one of claim 1-5 is proportionally added into into virus liquid, the parotid gland is made
Scorching viral lived vaccine stock solution；
Live mumps virus vaccine stock solution is pressed into target viral titre in vaccine semi-finished product and adds appropriate diluent preparing into vaccine
Semi-finished product, make each composition in vaccine freeze-drying protective agent final concentration of in vaccine semi-finished product：Sucrose 30-100g/L, glutamic acid
Sodium 0.5-15g/L, carbamide 0-10g/L, disodium hydrogen phosphate 1-20g/L, citric acid 0.2-10g/L；The diluent contains and this
The freeze drying protectant identical composition of invention, and the concentration of each composition and end of the freeze drying protectant of the present invention in vaccine semi-finished product
Concentration is consistent；The vaccine semi-finished product are the liquid vaccine before lyophilizing；The target viral titre is 5.8 ± 0.3lg CCID50/
ml；
Vaccine semi-finished product under the conditions of 2-8 DEG C, subpackage, lyophilizing.
9., according to the arbitrary described application of claim 7-8, the condition of lyophilizing is：Pre-freeze stage minimum temperature -45 DEG C~-48
DEG C, reach and maintained 2 hours after minimum temperature；- 35 DEG C of sublimation drying stage final temperature, the time for reaching final temperature is 17 little
When, vacuum pressure is controlled in 4-6Pa；28 DEG C of adsorption stripping and dry stage final temperature, vacuum is not controlled, and is run 8 hours.
10. the vaccine freeze-drying product that the arbitrary described application of claim 7-9 is prepared.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611267267.2A CN106668867B (en) | 2016-12-31 | 2016-12-31 | A kind of mumps vaccine freeze drying protectant without gelatin and human serum albumin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611267267.2A CN106668867B (en) | 2016-12-31 | 2016-12-31 | A kind of mumps vaccine freeze drying protectant without gelatin and human serum albumin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106668867A true CN106668867A (en) | 2017-05-17 |
| CN106668867B CN106668867B (en) | 2019-08-02 |
Family
ID=58850142
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611267267.2A Active CN106668867B (en) | 2016-12-31 | 2016-12-31 | A kind of mumps vaccine freeze drying protectant without gelatin and human serum albumin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106668867B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110101864A (en) * | 2019-05-31 | 2019-08-09 | 辽宁茂康源生物科技有限公司 | The protective agent of serum-free Antirabic Vaccine a kind of and its application |
| CN113144209A (en) * | 2021-01-19 | 2021-07-23 | 上海荣盛生物药业有限公司 | Rabies vaccine freeze-drying protective agent |
| CN115105604A (en) * | 2022-07-05 | 2022-09-27 | 吉林惠康生物药业有限公司 | Vaccine freeze-drying protective agent and freeze-drying method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104258404A (en) * | 2014-09-11 | 2015-01-07 | 长春长生生物科技股份有限公司 | Freeze-dried vaccine protective agent, freeze-dried varicella attenuated live vaccine and preparation methods of freeze-dried vaccine protective agent and freeze-dried varicella attenuated live vaccine |
| CN104548110A (en) * | 2015-01-10 | 2015-04-29 | 浙江卫信生物药业有限公司 | Mumps vaccine freeze-drying protective agent without gelatin and human serum albumin components |
| CN105999281A (en) * | 2016-07-26 | 2016-10-12 | 河南后羿生物工程股份有限公司 | Freeze-drying protective additive for poultry live viruses and preparation method and application thereof |
| CN106063933A (en) * | 2015-12-31 | 2016-11-02 | 武汉博沃生物科技有限公司 | General vaccines freeze drying protectant and application thereof |
-
2016
- 2016-12-31 CN CN201611267267.2A patent/CN106668867B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104258404A (en) * | 2014-09-11 | 2015-01-07 | 长春长生生物科技股份有限公司 | Freeze-dried vaccine protective agent, freeze-dried varicella attenuated live vaccine and preparation methods of freeze-dried vaccine protective agent and freeze-dried varicella attenuated live vaccine |
| CN104548110A (en) * | 2015-01-10 | 2015-04-29 | 浙江卫信生物药业有限公司 | Mumps vaccine freeze-drying protective agent without gelatin and human serum albumin components |
| CN106063933A (en) * | 2015-12-31 | 2016-11-02 | 武汉博沃生物科技有限公司 | General vaccines freeze drying protectant and application thereof |
| CN105999281A (en) * | 2016-07-26 | 2016-10-12 | 河南后羿生物工程股份有限公司 | Freeze-drying protective additive for poultry live viruses and preparation method and application thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110101864A (en) * | 2019-05-31 | 2019-08-09 | 辽宁茂康源生物科技有限公司 | The protective agent of serum-free Antirabic Vaccine a kind of and its application |
| CN113144209A (en) * | 2021-01-19 | 2021-07-23 | 上海荣盛生物药业有限公司 | Rabies vaccine freeze-drying protective agent |
| CN115105604A (en) * | 2022-07-05 | 2022-09-27 | 吉林惠康生物药业有限公司 | Vaccine freeze-drying protective agent and freeze-drying method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106668867B (en) | 2019-08-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105267971B (en) | A kind of vaccine freeze-drying protective agent without gelatin and human serum albumin | |
| CN101537186B (en) | Vaccine freeze-drying protective agent without gelatin | |
| CN102657870B (en) | Vaccine cryoprotectant without composition of gelatin and human albumin | |
| CN107412943A (en) | A kind of soluble microneedle patch and preparation method thereof | |
| CN103409375B (en) | Virus diluent for inoculating chick embryo and preparation method of virus diluent | |
| CN106237339B (en) | A kind of freeze dried vaccine heat resisting protective and its preparation method and application | |
| CN106668867B (en) | A kind of mumps vaccine freeze drying protectant without gelatin and human serum albumin | |
| CN106754762A (en) | A kind of antigen of encephalitis B live vaccine and preparation method and application | |
| CN101474410A (en) | Heat-resisting lyophilized protecting agent of live vaccine for animal and preparation method thereof | |
| CN104548110A (en) | Mumps vaccine freeze-drying protective agent without gelatin and human serum albumin components | |
| CN110917148B (en) | Blumea balsamifera combined attenuated live vaccine freeze-drying protective agent without gelatin and human blood albumin | |
| CN111588859B (en) | Freeze-drying protective agent and application thereof, freeze-dried seedling and preparation method thereof | |
| CN104367996A (en) | Method for producing swine pseudorabies live vaccine by using passage cell source, and product thereof | |
| CN102512685B (en) | A kind of vaccine protectant, measles encephalitis B combined vaccine and preparation method thereof | |
| CN105497904B (en) | A kind of pseudorabies virus vaccine heat-resisting lyophilized protecting agent and preparation method thereof | |
| CN115040656B (en) | Freeze-drying protective agent for rabies vaccine, application, vaccine and preparation method of vaccine | |
| CN114931648B (en) | Heat-resistant protective agent for combined live vaccine against porcine epidemic diarrhea and transmissible gastroenteritis, and preparation method and application thereof | |
| CN114652840A (en) | Freeze-drying protective agent for canine virus quadruple live vaccine and preparation method and application thereof | |
| CN110585439B (en) | Improved freeze-dried live attenuated hepatitis A vaccine stabilizer, vaccine semi-finished product, vaccine finished product and preparation method thereof | |
| CN1321694C (en) | Freezedrying type vaccine of AIDS recombined poxvirus with Ankara gene stock carrier, and preparation method | |
| CN113214388B (en) | Goose star virus egg yolk antibody and preparation method thereof | |
| CN115998853B (en) | Attenuated live vaccine for Japanese encephalitis and protective agent thereof | |
| CN104162149B (en) | Leukocyte interferon of pig freeze drying powder injection and preparation method thereof | |
| RU2522868C1 (en) | Method of production of vaccine against foot-and-mouth disease | |
| CN102309759B (en) | Poultry freeze-drying live vaccine heat-resisting protective agent and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |