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Multivalent pneumococcus capsular polysaccharide-protein conjugated composition and preparation method thereof

A technology of capsular polysaccharide and pneumococcus, which is applied in the field of microbiology and immunology, can solve the problems of limited protection range and lack of protection of serotype, and achieve the effect of reducing the number of vaccination needles, facilitating industrial large-scale production, and complete adsorption

Inactive Publication Date: 2014-03-12
SINOVAC RES & DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there is only 7-valent pneumococcal conjugate vaccine in my country Approved for marketing, it can only protect 7 valences, and the scope of protection is limited, unlike other 10-valent pneumococcal conjugate vaccines 13-valent pneumococcal conjugate vaccine Prevenar Its serotype combination is mainly designed for the epidemic situation in developed countries in Europe and the United States, and it lacks protection against the unique serotypes in my country

Method used

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  • Multivalent pneumococcus capsular polysaccharide-protein conjugated composition and preparation method thereof
  • Multivalent pneumococcus capsular polysaccharide-protein conjugated composition and preparation method thereof
  • Multivalent pneumococcus capsular polysaccharide-protein conjugated composition and preparation method thereof

Examples

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Embodiment 1

[0030] Example 1 Preparation of pneumococcal capsular polysaccharide stock solution

[0031] 1, 3, 4, 5, 6A, 6B, 9V, 14, 18C, 19A, 19F, 22F, 23F and 33F serotype Streptococcus pneumoniae strains were all obtained from China Institute for the Control of Pharmaceutical and Biological Products. The above strains were subcultured to establish the original seed bank, main seed bank and working seed bank. The generations were as follows: the original seed batch was the first generation, the main seed batch was the fourth generation, and the working seed batch was the eighth generation. From the opening of the working seed batch to the cultivation in the inoculated fermenter, the passage should not exceed 5 generations. The seeds of each generation use milk powder as a freeze-drying protective agent and freeze-dried for preservation. Take the working seeds and pick the strains by streaking on the soybean peptone solid medium, inoculate them in 5ml of liquid comprehensive medium, cul...

Embodiment 2

[0036] Example 2 Preparation of polyvalent pneumococcal capsular polysaccharide-protein conjugate composition

[0037] The 14 kinds of pneumococcal capsular polysaccharide stock solutions prepared in Example 1 were acidified and degraded with hydrochloric acid, and then sodium periodate was added to the polysaccharide for oxidation. After oxidation, the activation degree of the polysaccharide was detected, and then polysaccharide: protein = 1:2 The purified diphtheria toxoid CRM197 protein was added at a weight ratio of ~2:1 for binding, and sodium borohydride was added to terminate the binding reaction to prepare pneumococcal capsular polysaccharide conjugates, which were dialyzed by ultrafiltration to remove unbound proteins and polysaccharide, after purification, use a 0.2 μm filter to sterilize and filter, which is the pneumococcal capsular polysaccharide-protein monovalent conjugate.

[0038] The pneumococcal capsular polysaccharide-protein monovalent conjugates of the 1,...

Embodiment 3

[0039] Example 3 Preparation of multivalent pneumococcal capsular polysaccharide conjugate vaccine

[0040] The pneumococcal capsular polysaccharide-protein monovalent conjugates of 1, 3, 4, 5, 6A, 9V, 14, 18C, 19A, 19F, 22F, 23F and 33F serotypes prepared in Example 2 were prepared according to the final polysaccharide concentration of 4 μg / ml, the 6B conjugate was mixed according to the final polysaccharide concentration of 8 μg / ml, and aluminum phosphate adjuvant was added to prepare a 14-valent pneumococcal capsular polysaccharide conjugate vaccine with an aluminum content of 0.25 mg / ml.

[0041] The 14-valent pneumococcal capsular polysaccharide conjugate vaccine prepared above was made into a freeze-dried dosage form by low-temperature drying, and reconstituted with physiological saline before use.

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Abstract

The invention provides a multivalent pneumococcus capsular polysaccharide-protein conjugated composition and a preparation method thereof. The conjugated composition is formed through covalent linkages between pneumococcus capsular polysaccharides of 14 different serotypes and a carrier protein, wherein the 14 different serotypes are: 1, 3, 4, 5, 6A, 6B, 9V, 14, 18C, 19A, 19F, 22F, 23F, and 33F. The conjugated composition has a good absorption effect and stability, and has multiple immunogenicity and protective property aiming at the invasion of pneumococcus of the 14 serotypes; the effect of the conjugated composition is better than that of the low price and low quality pneumonia-treating compositions on the market, and moreover the immune response of the conjugated composition is higher than that of the composition that has not been combined. By using the multivalent pneumococcus capsular polysaccharide conjugate vaccine containing the conjugated composition, the inoculation times can be reduced, the immune procedure is simplified, diseases of humans and animals caused by the pneumococcus of the 14 serotypes can be effectively prevented, moreover the disease coverage is wider, and the immune effect is better.

Description

technical field [0001] The invention belongs to the field of microbiology and immunology, and in particular relates to a polyvalent pneumococcal capsular polysaccharide-protein conjugate composition and a preparation method thereof. Background technique [0002] Streptococcus pneumoniae can cause invasive diseases such as meningitis and pneumonia and non-invasive diseases such as otitis media. According to United Nations statistics, more than one million children under the age of two die each year from diseases caused by pneumococcus. At present, the main defenses against Streptococcus pneumoniae are polysaccharide vaccines and conjugate vaccines. The protective effect of the former in adults has been confirmed, but for the main susceptible population of Streptococcus pneumoniae, the protection in infants and young children is very weak . Because of its ability to induce effective protection and immune memory in infants and young children, conjugate vaccines have been incl...

Claims

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Application Information

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IPC IPC(8): A61K39/09A61K47/48C08B37/00A61P31/04
Inventor 韩星蔡芳王新立王治伟王见冬高强尹卫东
Owner SINOVAC RES & DEV
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