CN104548110A - Mumps vaccine freeze-drying protective agent without gelatin and human serum albumin components - Google Patents
Mumps vaccine freeze-drying protective agent without gelatin and human serum albumin components Download PDFInfo
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- CN104548110A CN104548110A CN201510021874.XA CN201510021874A CN104548110A CN 104548110 A CN104548110 A CN 104548110A CN 201510021874 A CN201510021874 A CN 201510021874A CN 104548110 A CN104548110 A CN 104548110A
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Abstract
The invention discloses a mumps vaccine freeze-drying protective agent without gelatin and human serum albumin components, and a formula and a preparation process of the mumps vaccine freeze-drying protective agent are provided. By adopting a technical scheme that the mumps vaccine freeze-drying protective agent is prepared from cane sugar, mycose, dextran, sodium glutamate, urea, arginine, a 199 culture medium, mannitol and sterile water for injection, a mumps live-attenuated vaccine freeze-drying preparation can avoid the defects of gelatin and human serum albumin, and the purposes of eliminating allergic reaction, removing hidden dangers of hepatitis infection and reducing cost can be achieved.
Description
Technical field
The present invention relates to a kind of freeze drying protectant, specifically referring to that the cheek Seedling of a kind of gelatin-free and human albumin's composition for making attenuated mumps vaccine, live lyophilized formulations freezes guarantor's agent.
Background technology
Freeze drying protectant, is called for short and freezes guarantor's agent, being when vaccine is made lyophilized formulations, being added on the protective agent for preventing vaccine from becoming because of lyophilization generation in vaccine; For making the freeze drying protectant of attenuated mumps vaccine, live lyophilized formulations, being called for short cheek Seedling and freezing guarantor's agent.
Prior art cheek Seedling freezes to be protected containing gelatin, human albumin's composition in agent, freezes protect attenuated mumps vaccine, live lyophilized formulations prepared by agent and be called for short existing cheek Seedling lyophilized formulations with prior art cheek Seedling; Biological medicine Technological research finds, the part of existing cheek Seedling lyophilized formulations is vaccinated crowd especially child, can produce strong anaphylaxis to gelatin wherein, human albumin's composition, or has the risk infecting hepatitis; Study carefully its mechanism, causing anaphylactoid is that the derivant of gelatin or gelatin can cause part vaccinate to produce the IgE antibody of anti-gelatin molecule, and gelatin both can cause cell mediated immune response, and acellular mediated immunity also can be caused to react; In addition, the endotoxin containing other strains in gelatin also can cause part vaccinate to produce exothermic reaction; In addition, causing part vaccinate to infect hepatitis is be derived from prior art cheek Seedling to freeze in the human albumin protected in agent and carry hepatitis source of disease, although human albumin derives from detect qualified healthy human blood, but by the limitation of existing detection technique level, the hepatitis source of disease of carrying in human albumin can produce recessive pollution to vaccine; Further, human albumin is in short supply expensive, and cost is high; Therefore, the problems and shortcomings that prior art exists anaphylaxis, infects hepatitis hidden danger, cost is high.
Summary of the invention
For above-mentioned prior art Problems existing with not enough, the present invention adopts the cheek Seedling prepared by sucrose, trehalose, dextran, sodium glutamate, carbamide, arginine, 199 culture medium, mannitol, sterilized water for injection to freeze the technical scheme of protecting agent, the cheek Seedling of a kind of gelatin-free and human albumin's composition is provided to freeze guarantor's agent, be intended to make attenuated mumps vaccine, live lyophilized formulations, avoid the drawback of gelatin, human albumin, reach and eliminate anaphylaxis, hepatitis of avoiding infection, the object that reduces costs.
The object of the present invention is achieved like this: the cheek Seedling of a kind of gelatin-free and human albumin's composition freezes guarantor's agent, comprises formula and preparation technology,
Described formula is,
Raw material 1: sucrose, pharmaceutical grade, 20 ~ 70;
Raw material 2: trehalose, pharmaceutical grade, 10 ~ 50;
Raw material 3: Dextran 40, pharmaceutical grade, 10 ~ 50;
Raw material 4: sodium glutamate, pharmaceutical grade, 5 ~ 15;
Raw material 5: carbamide, pharmaceutical grade, 2 ~ 8;
Raw material 6: arginine, pharmaceutical grade, 0.5 ~ 5;
Raw material 7:199 culture medium, 5 ~ 30;
Raw material 8: mannitol, pharmaceutical grade, 5 ~ 50;
Solvent: sterilized water for injection, surplus;
Formula unit is gram/1000 milliliters;
Described preparation technology is, mixing standardize solution;
Step one,
The solvent of the raw material 1 ~ 8 and 4/5 in formula is placed in beaker and is uniformly mixed dissolving, obtain mixed liquor;
Step 2,
Move in volumetric flask by described mixed liquor, carry out standardize solution with in 1/5 solvent injection capacity bottle remaining in formula, afterwards, cover the bottle stopper of volumetric flask, upset shakes up, and leaves standstill 3 ~ 5 minutes, obtains standardize solution liquid; Afterwards, with the hydrochloric acid of 20% pharmaceutical grade, the pH value of described standardize solution liquid is adjusted to 7.2 ~ 7.6; Obtain the thick liquid of standardize solution;
Step 3,
Move in container bottle by thick for described standardize solution liquid through 0.2 micron membrane filter aseptic filtration, capping, the cheek Seedling obtaining gelatin-free and human albumin's composition freezes protects agent goods.
Test
To freezing the safety of the cheek Seedling lyophilized formulations protecting the gelatin-free prepared of agent and human albumin's composition, effectiveness and stability with the cheek Seedling of gelatin-free of the present invention and human albumin's composition, protect existing cheek Seedling lyophilized formulations prepared by agent and carry out matched group test with freezing with prior art cheek Seedling;
Definition: in the scope of formula of the present invention, the cheek Seedling preparing the different gelatin-free of 3 groups of ratios and human albumin's composition freezes guarantor's agent, is called protective agent 1, protective agent 2, protective agent 3;
Take primary chick embryo cell as mumps virus cell culture prepared by culture matrix, add described protective agent 1 respectively, mumps virus harvest liquid that protective agent 2, protective agent 3 are produced, the gelatin-free obtained through subpackage lyophilizing and the cheek Seedling lyophilized formulations of human albumin's composition, be called sample 1, sample 2, sample 3;
The formula proportion of described protective agent 1 is, dextran 40,30; Sucrose, 50; Trehalose, 20; Sodium glutamate, 10; Carbamide, 5; Arginine, 2; Mannitol, 10; 199 culture medium, 10; Sterilized water for injection, surplus; Unit is gram/1000 milliliters;
The formula proportion of described protective agent 2 is, dextran 40,20; Sucrose, 50; Trehalose, 10; Sodium glutamate, 10; Carbamide, 5; Arginine, 2; Mannitol, 5; 199 culture medium, 10; Sterilized water for injection, surplus; Unit is gram/1000 milliliters;
The formula proportion of described protective agent 3 is, dextran 40,40; Sucrose, 50; Trehalose, 30; Sodium glutamate, 10; Carbamide, 5; Arginine, 2; Mannitol, 15; 199 culture medium, 10; Sterilized water for injection, surplus; Unit is gram/1000 milliliters;
Described prior art cheek Seedling freezes guarantor's agent and is called for short existing protective agent, and described existing protectant component is, gelatin, 1%; Sucrose, 5%; Sodium glutamate, 1%; Carbamide, 0.5%; Arginine, 0.2%; Human albumin, 1%;
Take primary chick embryo cell as mumps virus cell culture prepared by culture matrix, add the mumps virus harvest liquid that described existing protective agent is produced, the cheek Seedling lyophilized formulations obtained through subpackage lyophilizing is called available sample;
Carry out titre decline, 37 DEG C of stability and 2-8 DEG C of Detection of Stability before and after osmotic pressure, moisture, endotoxin content, lyophilizing respectively to sample 1, sample 2, sample 3 and available sample, result of the test is as follows:
A, osmotic pressure index Test
The method that sample 1, sample 2, sample 3 and available sample provide by " Chinese Pharmacopoeia " three annex V H is carried out osmometry respectively, and result is as follows:
Conclusion: the osmotic pressure of sample 1, sample 2, sample 3 is all in the scope of 400-500m0sm/kg, and well below available sample, the osmotic pressure index of sample 1, sample 2, sample 3 is better than available sample.
B, moisture are tested
The method that sample 1, sample 2, sample 3 and available sample provide by " Chinese Pharmacopoeia " three annex VII D is not carried out determination of water, and result is as follows:
Conclusion: the moisture of sample 1, sample 2, sample 3 and available sample is all less than 2%, meets the requirement that " Chinese Pharmacopoeia " moisture is less than 3%.
C, endotoxin content are tested
The method that sample 1, sample 2, sample 3 and available sample " Chinese Pharmacopoeia " three annex XII E provide is carried out endotoxin assay respectively, and result is as follows:
Conclusion: the endotoxin of sample 1, sample 2, sample 3 is all less than 5EU/ml, meets the requirement that " Chinese Pharmacopoeia " endotoxin content is less than 50EU/ml; Sample 1, sample 2, sample 3 are all better than available sample;
Titre dropping test before and after D, lyophilizing
Sample before sample 1, sample 2, sample 3 and available sample lyophilizing and after lyophilizing is carried out titration of virus respectively, and compare titre change, result is as follows:
Conclusion: the titre decline difference before and after sample 1, sample 2, sample 3 and available sample vaccine freeze-drying is all less than 1.0lg CCID
50/ ml, meets " Chinese Pharmacopoeia " requirement.
E, 37 DEG C of stability tests
Under sample 1, sample 2, sample 3 and available sample are placed in the condition of 37 DEG C, carried out titration of virus respectively at 0,7 day, compare titre change, result is as follows:
Conclusion: sample 1, sample 2, sample 3 and available sample are placed to 7 days under 37 DEG C of conditions, and virus titer drop-out value is less than 0.5lg CCID
50/ ml, the titre after simultaneously declining, still in the scope that " Chinese Pharmacopoeia " requires, meets " Chinese Pharmacopoeia " requirement.
F, 2 ~ 8 DEG C of stability tests
Under sample 1, sample 2, sample 3 and available sample are placed in the condition of 2 ~ 8 DEG C, carried out titration of virus respectively at 0,1,3,6,12,18,24 month, compare titre change, result is as follows:
Conclusion: sample 1, sample 2, sample 3 and available sample are being placed 24 months under 2-8 DEG C of condition, and virus titer, without obvious decline, meets " Chinese Pharmacopoeia " requirement.
Beneficial effect
To sum up test data proves, the cheek Seedling of gelatin-free of the present invention and human albumin's composition freezes the attenuated mumps vaccine, live lyophilized formulations protecting agent participation preparation and meets pharmacopoeial requirements, because the cheek Seedling of gelatin-free of the present invention and human albumin's composition freezes in guarantor's agent not containing gelatin, human albumin, avoid gelatin, human albumin to the harm of human body, enhance the safety of attenuated mumps vaccine, live lyophilized formulations, make attenuated mumps vaccine, live lyophilized formulations simultaneously, eliminate anaphylaxis, eliminate infection hepatitis hidden danger, reduce cost.
Above-mentioned, the present invention adopts the cheek Seedling prepared by sucrose, trehalose, dextran, sodium glutamate, carbamide, arginine, 199 culture medium, mannitol, sterilized water for injection to freeze the technical scheme of protecting agent, overcome prior art and there is anaphylaxis, infect hepatitis hidden danger, problems and shortcomings that cost is high, a kind of gelatin-free provided and the cheek Seedling of human albumin's composition freeze guarantor's agent, make attenuated mumps vaccine, live lyophilized formulations, avoid the drawback of gelatin, human albumin, reach and eliminate anaphylaxis, eliminate the object infecting hepatitis hidden danger, reduce costs.
Embodiment explanation
Freeze below by way of the cheek Seedling of specific embodiment to a kind of gelatin-free of the present invention and human albumin's composition and protect agent and be described in further detail, the cheek Seedling that those skilled in the art can obtain gelatin-free and human albumin's composition with reference to the formula of embodiment and preparation technology freezes guarantor's agent, but should not be construed as any limitation of the invention.
Detailed description of the invention
Embodiment
The name of an article: the cheek Seedling of gelatin-free and human albumin's composition freezes guarantor's agent
Formula: formula unit is gram/1000 milliliters;
Raw material 1: sucrose, pharmaceutical grade, 20 ~ 70;
Raw material 2: trehalose, pharmaceutical grade, 10 ~ 50;
Raw material 3: Dextran 40, pharmaceutical grade, 10 ~ 50;
Raw material 4: sodium glutamate, pharmaceutical grade, 5 ~ 15;
Raw material 5: carbamide, pharmaceutical grade, 2 ~ 8;
Raw material 6: arginine, pharmaceutical grade, 0.5 ~ 5;
Raw material 7:199 culture medium, 5 ~ 30;
Raw material 8: mannitol, pharmaceutical grade, 5 ~ 50;
Solvent: sterilized water for injection, surplus;
Preparation
Preparation condition, equipment
Ambient pressure a: atmospheric pressure; Ambient temperature: room temperature;
Prepare facility, equipment: sterilizing room; Beaker; Volumetric flask; Container bottle; 0.2 micron membrane filter;
Preparation technology: mixing standardize solution;
Step one,
The solvent of the raw material 1 ~ 8 and 4/5 in formula is placed in beaker and is uniformly mixed dissolving, obtain mixed liquor;
Step 2,
Move in volumetric flask by described mixed liquor, carry out standardize solution with in 1/5 solvent injection capacity bottle remaining in formula, afterwards, cover the bottle stopper of volumetric flask, upset shakes up, and leaves standstill 3 ~ 5 minutes, obtains standardize solution liquid; Afterwards, with the hydrochloric acid of 20% pharmaceutical grade, the pH value of described standardize solution liquid is adjusted to 7.2 ~ 7.6; Obtain the thick liquid of standardize solution;
Step 3,
Move in container bottle by thick for described standardize solution liquid through 0.2 micron membrane filter aseptic filtration, capping, the cheek Seedling obtaining gelatin-free and human albumin's composition freezes protects agent goods.
Claims (1)
1. the cheek Seedling of gelatin-free and human albumin's composition freezes guarantor's agent, comprises formula and preparation technology, it is characterized in that:
Described formula is,
Raw material 1: sucrose, pharmaceutical grade, 20 ~ 70;
Raw material 2: trehalose, pharmaceutical grade, 10 ~ 50;
Raw material 3: Dextran 40, pharmaceutical grade, 10 ~ 50;
Raw material 4: sodium glutamate, pharmaceutical grade, 5 ~ 15;
Raw material 5: carbamide, pharmaceutical grade, 2 ~ 8;
Raw material 6: arginine, pharmaceutical grade, 0.5 ~ 5;
Raw material 7:199 culture medium, 5 ~ 30;
Raw material 8: mannitol, pharmaceutical grade, 5 ~ 50;
Solvent: sterilized water for injection, surplus;
Formula unit is gram/1000 milliliters.
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| CN201510021874.XA CN104548110A (en) | 2015-01-10 | 2015-01-10 | Mumps vaccine freeze-drying protective agent without gelatin and human serum albumin components |
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| CN201510021874.XA CN104548110A (en) | 2015-01-10 | 2015-01-10 | Mumps vaccine freeze-drying protective agent without gelatin and human serum albumin components |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106668867A (en) * | 2016-12-31 | 2017-05-17 | 科兴（大连）疫苗技术有限公司 | Mumps vaccine freeze-drying protective additive free of gelatin and human albumin |
| 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 |
| CN110585439A (en) * | 2019-09-30 | 2019-12-20 | 长春生物制品研究所有限责任公司 | Improved freeze-dried live attenuated hepatitis A vaccine stabilizer, vaccine semi-finished product, vaccine finished product and preparation method thereof |
| CN110917148A (en) * | 2019-12-20 | 2020-03-27 | 北京民海生物科技有限公司 | Gelatin-free and human-blood-albumin-free gill wind combined attenuated live vaccine freeze-drying protective agent |
| JP2020520253A (en) * | 2017-05-17 | 2020-07-09 | ウニベルシダード ノヴァ デ リスボンＵｎｉｖｅｒｓｉｄａｄｅ Ｎｏｖａ Ｄｅ Ｌｉｓｂｏａ | Cryoprotectant and/or cryopreservative composition, method and uses thereof |
| CN115948311A (en) * | 2022-09-26 | 2023-04-11 | 天津科技大学 | Genetically engineered bacterium for producing 5-hydroxytryptophan and construction method and application thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106668867A (en) * | 2016-12-31 | 2017-05-17 | 科兴（大连）疫苗技术有限公司 | Mumps vaccine freeze-drying protective additive free of gelatin and human albumin |
| CN106668867B (en) * | 2016-12-31 | 2019-08-02 | 科兴（大连）疫苗技术有限公司 | A kind of mumps vaccine freeze drying protectant without gelatin and human serum albumin |
| JP2020520253A (en) * | 2017-05-17 | 2020-07-09 | ウニベルシダード ノヴァ デ リスボンＵｎｉｖｅｒｓｉｄａｄｅ Ｎｏｖａ Ｄｅ Ｌｉｓｂｏａ | Cryoprotectant and/or cryopreservative composition, method and uses thereof |
| 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 |
| CN110585439A (en) * | 2019-09-30 | 2019-12-20 | 长春生物制品研究所有限责任公司 | Improved freeze-dried live attenuated hepatitis A vaccine stabilizer, vaccine semi-finished product, vaccine finished product and preparation method thereof |
| CN110917148A (en) * | 2019-12-20 | 2020-03-27 | 北京民海生物科技有限公司 | Gelatin-free and human-blood-albumin-free gill wind combined attenuated live vaccine freeze-drying protective agent |
| CN110917148B (en) * | 2019-12-20 | 2022-08-23 | 北京民海生物科技有限公司 | Blumea balsamifera combined attenuated live vaccine freeze-drying protective agent without gelatin and human blood albumin |
| CN115948311A (en) * | 2022-09-26 | 2023-04-11 | 天津科技大学 | Genetically engineered bacterium for producing 5-hydroxytryptophan and construction method and application thereof |
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Application publication date: 20150429 |