Novel pneumococcal conjugate vaccine and preparation method thereof

A pneumococcal conjugate vaccine technology, which can be used in medical preparations containing active ingredients, antibacterial drugs, pharmaceutical formulations, etc., can solve problems such as restricting the development of pneumococcal conjugate vaccines and inconclusive PPV23 efficacy and effect.

Active Publication Date: 2010-07-28
复星安特金(成都)生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For more than 30 years, despite multiple studies, the potency and effectiveness of PPV23 in children and adults re

Method used

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  • Novel pneumococcal conjugate vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Example 1 Fermentation culture of pneumococcus and preparation of capsular polysaccharide

[0078] Open the cryopreserved pneumococcal freeze-dried strains, redissolve with physiological sodium chloride solution, inoculate into 5-10% sheep blood agar slant medium, place in 5-8% CO 2 In the environment, cultivate at 35-37°C for 16-24 hours. Transfer to pneumococcal liquid basal medium, and cultivate at 35-37°C for 10-18 hours. Continue to transfer to pneumococcal liquid basal medium for enrichment culture, and cultivate at 35-37°C for 10-18 hours. Inoculate the enriched pneumococcal liquid culture into the fermenter with liquid basal medium according to the inoculum amount of 5-10%, stir and ventilate, cultivate at 35-37°C for 8-12 hours, and control the pH at 6.0 ~7.4. During the fermentation period, the concentration of bacteria was monitored, and the purity of the culture and the morphological characteristics of the bacteria were checked under a microscope. At the...

Embodiment 2

[0082] Example 2 Preparation of Pneumococcal Capsular Oligosaccharides

[0083] The purified polysaccharide is dissolved in 0.2-1.0M acetic acid solution to a final concentration of 1-4mg / ml, and immersed in an oil bath or water bath at 70-100°C for 6-40 hours for hydrolysis. After hydrolysis, the pH was adjusted to 7.0, and dialyzed at 2-8°C. Purified by Sepharose 4FF chromatography column, eluted with 15mM NaCl solution, and collected oligosaccharides. Purified oligosaccharides were obtained by 0.22μm sterile filtration and stored below -20°C in liquid or lyophilized form. Purified oligosaccharides were tested according to the methods and standards stipulated in the pneumococcal conjugate vaccine manufacturing and testing regulations officially promulgated by the World Health Organization (WHO Technical Report Series, No.927, 2005).

[0084] Prepare pneumococcal serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20 according to the ...

Embodiment 3

[0085] Example 3 Preparation of meningococcal capsular saccharide and Haemophilus influenzae type b capsular saccharide

[0086] According to the method of Chinese patent 200710007045.1, the capsular polysaccharides of group A, group C, group W135 and group Y meningococcus and the capsular polysaccharide of Haemophilus influenzae type b were prepared.

[0087] Then according to the method of Example 2 of the present invention, the capsular polysaccharides of group A, group C, group W135 and group Y meningococcus and the capsular oligosaccharide of Haemophilus influenzae type b were prepared.

[0088] Manufacture and test regulations (WHO Technical Report Series, No.658, 1980) and Haemophilus influenzae type b conjugate vaccines (WHO Technical Report Series, No.897, 2000) stipulated methods and standards to test the purified polysaccharides and oligosaccharides.

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Abstract

The invention relates to a novel pneumococcal conjugate vaccine and a preparation method thereof. The novel pneumococcal conjugate vaccine comprises a dual-valent serum type pneumococcal capsular sugar-protein combination and/or a multi-valent serum type pneumococcal capsular sugar-protein combination. The dual-valent serum type capsular sugar-protein combination is a combination formed by combining two different serum types of capsular sugar with a protein carrier through chemical bonds, and the two different serum types of capsular sugar form links in the structure according to the combination with the protein carrier through the chemical bonds. The multi-valent serum type capsular sugar-protein combination is a combination formed by combining two different serum types of capsular sugar with a protein carrier through chemical bonds, and the two different serum types of capsular sugar form links in the structure according to the combination with the protein carrier through the chemical bonds. In addition, the invention also provides a method of preparing the bacterial capsular sugar-protein conjugate vaccine.

Description

technical field [0001] The present invention relates to a new pneumococcal conjugate vaccine. Specifically, the present invention relates to a new pneumococcal conjugate vaccine in which capsular saccharides of multiple serotypes of pneumococcus are combined with protein carriers. After capsular saccharides from different serotypes are chemically combined with protein carriers, the The valent serotype capsular saccharide-protein conjugate and / or the multivalent serotype capsular saccharide-protein conjugate form are present in the vaccine. The invention also relates to the preparation method of the novel pneumococcal conjugate vaccine and the preparation method of the novel bacterial capsule glycoprotein conjugate vaccine. Background technique [0002] The disease caused by Streptococcus pneumoniae is the most important cause of morbidity and mortality among children in the world, especially in developing countries. The disease burden of pneumococcus mainly comes from child...

Claims

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Application Information

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IPC IPC(8): A61K39/09A61K39/116A61K39/095A61K39/102A61K39/112A61K39/085A61K39/39A61P31/04
CPCY02A50/30
Inventor 郑玉清
Owner 复星安特金(成都)生物制药有限公司
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