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Method for producing novel freeze-dried live attenuated varicella vaccine

A technology of a live attenuated vaccine and a production method, which is applied in the production field of a new type of freeze-dried live attenuated varicella vaccine, can solve problems such as restrictions on production methods, and achieve the effects of rapid preparation, improved safety and stability, and high purity

Inactive Publication Date: 2014-12-10
BEIJING HEKANGYUAN BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Common human serum albumin is mostly extracted from human plasma. This production method is not only limited by the supply of plasma, but also may contain dangerous infectious disease pathogens, such as hepatitis virus, HIV virus, etc., which makes it difficult for people to use plasma There are huge concerns about extracting human serum albumin from

Method used

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  • Method for producing novel freeze-dried live attenuated varicella vaccine
  • Method for producing novel freeze-dried live attenuated varicella vaccine
  • Method for producing novel freeze-dried live attenuated varicella vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] A production method of a novel freeze-dried live attenuated varicella vaccine, the production method comprising the following steps:

[0028] 1) Recovery and subculture of human diploid cell lines;

[0029] Resuscitate MRC-5 cells: Take out the frozen MRC-5 cells from liquid nitrogen, thaw them immediately in a 37°C water bath, centrifuge at 1000rpm for 5min, discard the upper layer of frozen storage solution, add fresh culture medium, mix gently and set aside for 37 Cultivate at ℃ for 2-4 days until the cells cover a single layer; wherein, the culture medium is MEM solution containing 10% calf serum and 1% glutamine, and no antibiotics are added during the vaccine production process.

[0030] Digestion and subculture: When the cells grow into a uniform and dense monolayer, pour off the culture medium in the culture bottle, add trypsin digestion solution to digest until the cell attachment is loose, the cell edge rolls up and the interval between cells increases, then p...

Embodiment 2

[0050] This embodiment is a preferred solution on the basis of embodiment 1. The quality of the raw materials used is the same as that of embodiment 1, and the same part as embodiment 1 will not be repeated. Please refer to embodiment 1.

[0051] The raw materials of the lyoprotectant include sorbitol, recombinant human serum albumin, sucrose, dextran, trehalose, arginine, sodium glutamate and urea, and the ratio of parts by weight of each raw material is:

[0052]

[0053] The lyoprotectant is the 199 comprehensive culture medium containing the above-mentioned raw materials.

[0054] The recombinant human serum albumin described in this example is yeast-expressed recombinant human serum albumin Catalog No. PRO-332 produced by Prospec Company of Israel.

[0055] The microcarriers described in this example are Cytodex series microcarriers produced by General Electric Company of the United States; the stirred bioreactor produced by NBS Company of the United States is used in ...

Embodiment 3

[0058] This embodiment is a preferred solution on the basis of embodiment 1. The quality of the raw materials used is the same as that of embodiment 1, and the same part as embodiment 1 will not be repeated. Please refer to embodiment 1.

[0059] The raw materials of the lyoprotectant include sorbitol, recombinant human serum albumin, sucrose, dextran, trehalose, arginine, sodium glutamate and urea, and the ratio of parts by weight of each raw material is:

[0060]

[0061]

[0062] The lyoprotectant is the 199 comprehensive culture medium containing the above-mentioned raw materials.

[0063] The microcarriers described in this example are Fibra-Cel Disks series microcarriers produced by NBS Company of the United States; the packed bed bioreactor produced by Biou Company of Switzerland is used in this example.

[0064] Please refer to Table 1 for the titer, moisture analysis and stability test results of the varicella vaccine.

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Abstract

The invention discloses a method for producing a novel freeze-dried live attenuated varicella vaccine. The method is characterized in that a human diploid cell is used as a culture medium, a reactor culture process is applied to inoculate a varicella OKa strain, and an invented freeze-drying protective additive containing sorbitol and recombinant human serum albumin is used to produce the freeze-dried live attenuated varicella vaccine. By applying the reactor culture technology, the production cost is reduced, the defects of low yield, large production occupied area, high labor intensity and the like of a static culturing process can be overcome, the yield of vaccines can be increased, and the virus titer stability can be improved; furthermore, the protective additive containing sorbitol and recombinant human serum albumin is used and does not contain gelatin, so that animal derived ingredients can be removed, the content of toxin in the vaccine can be remarkably reduced, the stimulation and hazard of the vaccine on a human body can be greatly reduced, and the safety and stability of the vaccine can be improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a production method of a novel freeze-dried live attenuated varicella vaccine. Background technique [0002] Chickenpox is an acute respiratory infectious disease caused by the varicella-zoster virus. It is highly contagious and commonly occurs in children. Its main clinical features are fever accompanied by itchy rashes, macules, papules, and vesicular eruptions all over the body. Because most of the symptoms of chickenpox in childhood are mild, people used to not pay attention to it. However, if the chickenpox is not treated properly, it will be complicated by serious diseases such as pneumonia and encephalitis, and even death. Shingles occurs. The incidence and harm of chickenpox among children in our country are becoming increasingly prominent. Vaccination of varicella vaccine is an effective method to control the infection and epidemic of varicella. How to further increase the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/19A61K39/25A61K47/26A61K47/42A61P31/22
Inventor 高辉
Owner BEIJING HEKANGYUAN BIOLOGICAL SCI & TECH
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