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Vaccine freeze-drying protective agent without gelatin

A freeze-drying protective agent and freeze-drying technology, which can be used in freeze-dried transportation, medical preparations containing active ingredients, antiviral agents, etc., can solve the problems of poor vaccine protection effect, decreased vaccine stability, and prone to failure. Achieve good protection effect, improve stability and improve safety

Active Publication Date: 2009-09-23
CHANGCHUN BCHT BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the gelatin-free vaccine protective agent in the prior art reduces the irritation to the human body, the protective effect on the vaccine is not as good as the commonly used protective agent containing gelatin, which makes the stability of the vaccine during freeze-drying and storage. drop, prone to failure

Method used

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  • Vaccine freeze-drying protective agent without gelatin
  • Vaccine freeze-drying protective agent without gelatin
  • Vaccine freeze-drying protective agent without gelatin

Examples

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Effect test

preparation example Construction

[0017] In a preferred embodiment, the present invention provides a kind of preparation method of freeze-dried varicella vaccine, comprises the following steps:

[0018] (1) take the varicella virus Oka strain as the virus seed, and the MRC-5 strain human diploid cells as the culture substrate;

[0019] (2) amplify and passage MRC-5 strain human diploid cells with culture medium, wherein the cell passage ratio is 1:2-1:4, and culture at 37°C for 3-5 days;

[0020] (3) After the human diploid cells of the MRC-5 strain grow into uniform and dense monolayer cells, replace the medium maintenance solution and add virus species to infect the cells;

[0021] (4) When more than 75% of the typical lesions appear in the human diploid cells of the MRC-5 strain, the original maintenance solution is poured out, and the cell surface is washed with a washing solution equivalent to 2-3 times the volume of the maintenance solution;

[0022] (5) Harvest the cell culture by adding the vaccine li...

preparation Embodiment 1

[0031] (1) Take human diploid cell MRC-5 at a ratio of 1:2, and use MEM cell culture medium with pH 7.2 and supplemented with 10% calf serum as the medium for cell subculture.

[0032] (2) At 37°C, culture for 3 days and amplify within 33 passages.

[0033] (3) After the MRC-5 cells form a uniform and dense 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. The 0ka strain of varicella virus was added to infect the cells.

[0034] (4) When more than 75% of the MRC-5 cells have typical lesions, 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.

[0035] (5) Add the vaccine solution to harvest the cell culture; the vaccine solution is: containing 10g / l human serum albumin, 40g / l sucrose, 15g / l trehalose, 50g / l dext...

preparation Embodiment 2

[0039] The steps of this embodiment are similar to Preparation Example 1, the difference is:

[0040] The maintenance solution used in step (3) is MEM medium with pH 7.4 and containing 2% calf serum, and the ratio of virus seed to cell inoculation is 1:90.

[0041] In step (4), the cell surface is washed with PBS buffer equivalent to 2 times the volume of the original maintenance solution, and the bovine serum is washed away.

[0042] The vaccine liquid used in step (5) is: containing 20g / l human serum albumin, 50g / l sucrose, 10g / l trehalose, 20g / l dextran 70, 10g / l sodium glutamate, 9g / l urea and 199 comprehensive medium with 0.5g / l arginine.

[0043]The final concentration of mannitol added in the stock solution in step (7) is 15g / l.

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Abstract

The invention relates to a composition used as a vaccine freeze-drying protective agent, comprising the components with initial concentration in the freeze-drying stock solution as follows: 30-100g / l sucrose, 10-30g / l trehalose, 10-50g / l dextran, 6-12g / l sodium glutamate, 3-9g / l urea and 0.5-2g / l arginine, and no gelatin. The invention further provides a method for preparing freeze-drying vaccines by using the freeze-drying protective agent and application of the freeze-drying protective agent for improving the stability of freeze-drying vaccines.

Description

technical field [0001] The invention belongs to the technical field of vaccine production technology, and in particular relates to the formulation and application method of a gelatin-free vaccine protective agent. Background technique [0002] At present, the application of preventive vaccines has effectively controlled common epidemic infectious diseases and effectively reduced the morbidity and mortality of these infectious diseases. However, in the process of storage, transportation and clinical use, a series of problems such as poor stability, short validity period and high adverse reaction rate continue to appear in the vaccine products produced by the existing production process. With the wide application of vaccines and the rapid development of molecular biology and cell biology, studies have found that many problems are directly related to the protective agents and adjuvants of vaccine products, especially freeze-dried vaccine products need to use lyoprotectants, Th...

Claims

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Application Information

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IPC IPC(8): A61K47/42A61K47/36A61K47/26A61K47/18A61K47/10A61K39/25A61P31/22
CPCA61K47/36A61K9/19A61K47/42A61K47/26A61K39/00A61K47/18A61P31/22
Inventor 朱昌林沈艳杰祝洪敢李海泉孙会来徐艳君张喆王晓丽
Owner CHANGCHUN BCHT BIOTECH
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