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Method for preparation of freeze-dried chickenpox vaccine

A freeze-drying and chickenpox technology, applied in the field of preparation of chickenpox vaccine, can solve problems such as poor vaccine stability, and achieve the effect of good vaccine stability

Inactive Publication Date: 2007-03-14
长春百克生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The invention provides a preparation method of freeze-dried varicella vaccine to solve the problem of poor stability of current vaccines

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Take human diploid cell MRC-5 at a ratio of 1:2, use MEM cell culture medium with pH 7.2 and supplemented with 10% calf serum as the medium for cell subculture, and culture for 3 days at 37°C to expand and subculture, 33 Within one generation; after the MRC-5 cells form a uniform and dense cell monolayer, replace the MEM medium maintenance medium with pH 7.2 and containing 2% calf serum, and inoculate the culture bottle at a ratio of 1:60 virus species and cells Add varicella virus Oka strain to infect the cells; when more than 75% of typical lesions appear on the MRC-5 cells, pour off the original maintenance solution, wash the cell surface with Earle's solution equivalent to more than twice the volume of the original maintenance solution, and wash away the bovine serum ; add 199 comprehensive medium vaccine solution containing 0.3% human albumin and 10% sucrose, and harvest the cell culture; freeze and thaw the cell culture below -70°C, 20KHz ultrasonically break the c...

Embodiment 2

[0014] Take human diploid cell MRC-5 at a ratio of 1:4, use MEM cell culture medium with pH 7.6 and supplemented with 12% calf serum as the medium for cell subculture, and culture for 5 days at 37°C for expansion and subculture, 33 Within one generation; after the MRC-5 cells form a uniform and dense cell monolayer, replace the MEM medium maintenance medium with pH 7.4 and containing 5% calf serum, and inoculate the virus species and cells at a ratio of 1:100 in the culture bottle Add varicella virus Oka strain to infect the cells; when more than 75% of typical lesions appear on the MRC-5 cells, pour off the original maintenance solution, wash the cell surface with PBS buffer equivalent to 2 times the volume of the original maintenance solution, and wash away the bovine serum; Add PBS buffer vaccine solution containing 2% human albumin and 3% sucrose, and harvest the cell culture; freeze and thaw the cell culture below -70°C, break the cells with 20KHz ultrasound, and clarify a...

Embodiment 3

[0016] Take human diploid cell MRC-5 at a ratio of 1:3, use MEM cell culture medium with pH 7.4 and supplemented with 15% calf serum as the medium for cell subculture, and culture for 4 days at 37°C for expansion and subculture, 33 Within one generation; after the MRC-5 cells form a uniform and dense cell monolayer, replace the MEM medium maintenance medium with pH 7.6 and containing 3.5% calf serum, and inoculate the culture bottle at a ratio of 1:120 virus seeds and cells Add varicella virus Oka strain to infect the cells; when more than 75% of typical lesions appear on the MRC-5 cells, pour off the original maintenance solution, wash the cell surface with Earle's solution 3 times the volume of the original maintenance solution, and wash away the bovine serum; add 1 The 199 comprehensive culture medium vaccine solution containing % human albumin and 8% sucrose is used to harvest the cell culture; the cell culture is frozen and thawed below -70°C, the cells are ultrasonically ...

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Abstract

The invention relates to the preparative way of a kind of freeze-drying varicella vaccine, which has something to do with the biotechnology. The main making procedures include the following steps: the amplification and passage of Varicellavirus strain Oka to the MRC-5 strain in human's diploid cells; the harvest of the vaccine liquids; freeze-drying. The main property of the vaccine is that it adopts 0.5-1.2%(g / ml) gelatine, 0.6-1.2%(g / ml) glutamic sodium, 0.3-0.9%(g / ml) carbamide and 0.05-0.2%(g / ml) arginine as the freeze-drying protective agents, the previous concentrations stand for the final concentration of the protective agents. The good quality of the vaccine lies in that: the freeze-drying protective agents are developed by the producers themselves to make sure the stability of the vaccine. The result of storage stability test at 2-8DEG C shows that: there's no evident difference between the samples that stored for 18 months and the new-produced samples.

Description

technical field [0001] The invention belongs to the technical field of vaccine production and preparation technology, and in particular relates to a preparation method of varicella vaccine. Background technique [0002] Varicella-zoster virus (VZV) is the causative agent of chickenpox and shingles. Chickenpox is the manifestation of the primary infection of the virus, mainly infecting children, characterized by the appearance of herpes on the skin and mucous membranes. Immunity for life after the disease, generally no second infection; Caused by activation of the varicella-zoster virus, mainly in adults, especially those with cancer and the elderly who are receiving immunosuppressive therapy, characterized by inflammation of the dorsal roots and ganglia of the spinal nerves that innervate the Herpes on the skin, pain or nerve pain. Both chickenpox and shingles can have serious complications. [0003] VZV is a member of the Alphaherpesvirinae subfamily of the Herpesvirida...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/275A61K9/19A61P31/20
Inventor 朱昌林魏巍孙会来孔维王忠诚郭丽姝万铁军曾智姜春来
Owner 长春百克生物科技有限公司
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