Preparation method of pneumococcal conjugate combination vaccine

A pneumococcal and combined vaccine technology, applied in vaccines, multivalent vaccines, antibacterial drugs, etc., can solve problems such as unsatisfactory effects

Pending Publication Date: 2016-11-16
BEIJING ZHIFEI LVZHU BIOPHARM +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Kossaczka etc. prepared typhoid conjugate vaccine by this method, but the human experiment data proves that the effect is not satisfactory (KossaczkaZuzana, FY Lin, VAHo, ...

Method used

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  • Preparation method of pneumococcal conjugate combination vaccine
  • Preparation method of pneumococcal conjugate combination vaccine
  • Preparation method of pneumococcal conjugate combination vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1: Pneumococcus bacterial culture

[0057] After the strains are turned on, transfer them to Columbia blood agar medium, culture them at 37°C, recover and grow colonies, transfer them to a small amount of improved pancreatic soybean liquid medium, cultivate them for 8-12 hours, and transfer them to the second-generation improved pancreatic soybean liquid medium. Soybean strains are cultivated in liquid medium, and then transferred to a fermenter containing 35000ml of liquid medium for cultivation, during which the pH value is maintained at about 7.0, and the bacteria are inactivated at the end of the logarithmic growth phase.

Embodiment 2

[0058] Example 2: Purification of pneumococcal capsular polysaccharide

[0059] After the culture supernatant was centrifuged, concentrated by 100K membrane ultrafiltration, the buffer was replaced with 0.01mol / L phosphate buffer, the pH value was adjusted to 6.8-7.4, and cetyltrimethylammonium bromide was added at a final concentration of 1%. (CTAB), stir well and leave overnight. Collect the precipitate by centrifugation (collect supernatant for 7F and 14 types), add 0.25mol / L NaCl solution to dissociate CTAB (use 0.35mol / L NaCl solution for type 3), add 0.5% NaI, stir well, and collect supernatant by centrifugation , with ultrafiltration membrane ultrafiltration dialysis. After hydroxyapatite chromatography, the target polysaccharide is collected, precipitated with 75% ethanol, washed with ethanol and acetone, and dried to obtain purified polysaccharide.

Embodiment 3

[0060] Example 3: Test results of purified polysaccharides

[0061] The 24 pneumococcal capsular polysaccharides were purified separately, with 3 batches for each type, and the quality inspection results of each batch are shown in Table 3:

[0062]Table 3. Quality detection results of pneumococcal capsular polysaccharide

[0063]

[0064]

[0065]

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Abstract

The invention relates to pneumococcal conjugate combinations. High-immune conjugates are prepared with selected combination methods suitable for different polysaccharide types and constitute one combination which is the pneumococcal conjugate combination with high immunogenicity, and the combination is used for preventing infectious diseases caused by pneumococcus. The combination vaccine is prepared from pneumococcal conjugate combinations selected from pneumococcus serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33 F.

Description

Technical field: [0001] The present invention relates to a combination of pneumococcal conjugates. High immune conjugates are prepared by selecting a combination method suitable for different types of polysaccharides, and they are combined into a combination to become a highly immunogenic pneumococcal conjugate combination for preventing pneumonia. Infectious diseases caused by bacteria. Background technique: [0002] Streptococcus pneumoniae is the most important pathogen causing community acquired pneumonia (CAP) in all age groups in developing and developed countries, and also the main pathogen causing otitis media, pneumonia, meningitis and bacteremia. Worldwide, the annual death toll caused by Streptococcus pneumoniae infection accounts for 9% of the total death toll, more than 30,000 cases of meningitis, more than 630,000 cases of bacteremia, 14 million cases of otitis media, and pneumonia caused by Streptococcus pneumoniae. 20 million cases. [0003] Biological prod...

Claims

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Application Information

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IPC IPC(8): A61K39/116A61K39/09A61P31/04
CPCA61K39/09A61K2039/55505A61K2039/6037A61K2039/6068A61K2039/70
Inventor 杜琳朱卫华林彦彬王瑞峰胡小华宗向坤徐慧娴胡月凤
Owner BEIJING ZHIFEI LVZHU BIOPHARM
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