Method for preparing streptococcus pneumoniae capsular polysaccharide

A technology for Streptococcus pneumoniae and capsular polysaccharide, which can be used in microorganism-based methods, biochemical equipment and methods, antibacterial drugs, etc., and can solve the problems of many laboratory methods and difficult industrialization.

Active Publication Date: 2014-06-04
TASLY BIOPHARMACEUTICALS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Pneumococcus capsular polysaccharide is an effective component of pneumonia vaccine. There are many existing preparation methods, but there are many laboratory methods, which are difficult to apply to ind

Method used

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  • Method for preparing streptococcus pneumoniae capsular polysaccharide
  • Method for preparing streptococcus pneumoniae capsular polysaccharide
  • Method for preparing streptococcus pneumoniae capsular polysaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Example 1: The cultivation of Streptococcus pneumoniae 19F and the extraction of Pn19F-Ps

[0080] 1. Culture of Streptococcus pneumoniae 19F

[0081] Strains were obtained from the American Type Culture Collection (ATCC 6319).

[0082] The conventional culture method of Streptococcus pneumoniae is well known in the art, and the liquid medium that can be used includes: TSB, CY, BHI, Hoeprich's medium and its improved formula etc.

[0083] Take Streptococcus pneumoniae type 19F working seeds from the -70°C refrigerator, inoculate them on nutrient agar plates containing 10% defibrated sheep blood, at 37°C ± 1°C, 5% CO 2 Cultivate in the incubator for 18-24h. Verify the absence of contaminants.

[0084] Scrape off the above culture, inoculate in several 1L (500ml medium capacity) Erlenmeyer flasks, at 37°C±2°C, 5% CO 2 Cultivate in the incubator for 14-20h, and monitor the cell density (600nm) during this process. When the cell density reaches 2.0-2.5, use plate marki...

Embodiment 2

[0095] Embodiment 2: the cultivation of Streptococcus pneumoniae 9V and the extraction of Pn9V-Ps

[0096]1. Culture of Streptococcus pneumoniae 9V

[0097] Strains were obtained from the American Type Culture Collection (ATCC 6309).

[0098] The conventional culture method of Streptococcus pneumoniae is well known in the art, and the liquid medium that can be used includes: TSB, CY, BHI, Hoeprich's medium and its improved formula etc.

[0099] Take Streptococcus pneumoniae type 9V working seeds from the -70°C refrigerator and inoculate them on nutrient agar plates containing 10% defibrated sheep blood, at 36°C ± 1°C, 5% CO 2 Cultivate in the incubator for 18-24h. Verify the absence of contaminants.

[0100] Scrape off the above culture, inoculate in several 1L (500ml medium capacity) Erlenmeyer flasks, at 36°C±2°C, 5% CO 2 Cultivate in the incubator for 14-20h, and monitor the cell density (OD600nm) during this process. When the cell density reaches 2.0-2.5, use plate ma...

Embodiment 3

[0108] Embodiment 3: the cultivation of Streptococcus pneumoniae type 5 and the extraction of Pn5-Ps

[0109] 1. Culture of Streptococcus pneumoniae type 5

[0110] Strains were obtained from the American Type Culture Collection (ATCC 6305).

[0111] The conventional culture method of Streptococcus pneumoniae is well known in the art, and the liquid medium that can be used includes: TSB, CY, BHI, Hoeprich's medium and its improved formula etc.

[0112] Take the working seeds of Streptococcus pneumoniae type 5 from the -70°C refrigerator, inoculate them on nutrient agar plates containing 10% defibrated sheep blood, and store them at 37°C±2°C, 5% CO 2 Cultivate in the incubator for 18-24h. Verify the absence of contaminants.

[0113] Scrape off the above culture, inoculate several 1L (500ml culture medium) Erlenmeyer flasks at 37°C ± 1°C, 5% CO 2 Cultivate in the incubator for 14-20h, and monitor the cell density (600nm) during this process. When the cell density reaches 2....

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Abstract

The invention relates to a method for preparing streptococcus pneumoniae capsular polysaccharide. The method comprises the following steps: 1, culturing streptococcus pneumoniae; 2, inactivating; 3, extracting streptococcus pneumoniae capsular polysaccharide; 4, refining streptococcus pneumoniae capsular polysaccharide.

Description

Technical field: [0001] The invention relates to the preparation of vaccines, in particular to a preparation method of Streptococcus pneumoniae capsular polysaccharide. Background technique: [0002] Streptococcus pneumoniae, also known as Diplococcus pneumoniae, belongs to the genus Streptococcus. Streptococcus pneumoniae can cause a variety of diseases, mainly pneumonia, meningitis, bacteremia, otitis media and so on. This bacterium is an opportunistic pathogenic bacterium, which can form a carrier state in the oral cavity and nasopharyngeal cavity of normal people, and cause diseases when the body's resistance declines, especially the elderly, children under 2 years old, and those with immunodeficiency or low immunity crowd. [0003] It is conservatively estimated that Streptococcus pneumoniae causes more than 1 million deaths of children under 5 years old every year in developing countries, and about 20% to 25% of the deaths are in this age group, even in developed cou...

Claims

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Application Information

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IPC IPC(8): C08B37/00C12P19/04A61K39/09A61P31/04C12R1/46
Inventor 金永杰孙倩李军强赵秋敏李亚冰刘贺军曹小丹李静李剑
Owner TASLY BIOPHARMACEUTICALS CO LTD
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