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Pneumococcal polysaccharide and protein conjugated vaccine and preparation method thereof

A pneumococcal polysaccharide and pneumococcal technology, applied in the field of immunology, can solve the problems of increasing the amount of carrier protein, prone to immune tolerance, and reduced immunogenicity

Active Publication Date: 2014-07-02
CANSINO BIOLOGICS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore polysaccharide vaccine has following problem: (1) can only produce weak immune response in young animal or infant body, even does not produce immune response, and immune response increases with age; (2) produces the antibody of low affinity; 3) It only produces a short-term immune response, and does not have the immune memory and immune enhancement effect during repeated vaccination; (4) It is easy to produce immune tolerance; (5) Common adjuvants are not easy to play an immune enhancement effect on this antigen
[0010] Although the 13-valent pneumococcal conjugate vaccine has been marketed, the immune effect of several serotypes is lower than that of other serotypes, such as type 3
And with the increase of different serotypes and repeated use of the same carrier protein, the amount of carrier protein increases, but its immunogenicity decreases

Method used

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  • Pneumococcal polysaccharide and protein conjugated vaccine and preparation method thereof
  • Pneumococcal polysaccharide and protein conjugated vaccine and preparation method thereof
  • Pneumococcal polysaccharide and protein conjugated vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Preparation of a 7F-type capsular polysaccharide and two different carrier protein conjugates

[0050] Dissolve 5g of the lung chain 7F type polysaccharide in 1L of sodium acetate buffer (50mM pH4.5), at room temperature, stir for 20 minutes with a magnetic stirrer with a magnetic sub, in order to fully dissolve the polysaccharide, add 0.2g NaIO 4 , and react overnight at room temperature, so that the adjacent dihydroxyl groups on the polysaccharide are oxidized into aldehyde groups. Perform 20 equal-volume ultrafiltration changes with purified water, and remove the remaining NaIO in the reaction 4 And the small molecules generated in the reaction process were filtered off to obtain an aqueous solution in which the 7F activated polysaccharide with an activation degree of 6.2 was introduced; and the activated polysaccharide could be concentrated and freeze-dried. The molecular size of the activated polysaccharide was determined using a gel filtration chromatog...

Embodiment 2

[0055] Example 2 Preparation of a type 3 capsular polysaccharide and two different carrier protein conjugates

[0056] Dissolve 2g of lung chain type 3 polysaccharide in 1L sodium phosphate buffer (50mM pH7.2), and stir for 20 minutes with a magnetic stirrer with a magnet at room temperature to fully dissolve the polysaccharide, add 0.2g NaIO 4 , and react overnight at room temperature, so that the adjacent dihydroxyl groups on the polysaccharide are oxidized into aldehyde groups. Perform 20 equal-volume ultrafiltration changes with purified water, and remove the remaining NaIO in the reaction 4 and the small molecular substances generated during the reaction were filtered off to obtain an aqueous solution in which the type 3 activated polysaccharide with an activation degree of 9.1 was introduced; and the activated polysaccharide was concentrated and freeze-dried. The molecular size of the activated polysaccharide was determined using a gel filtration chromatography column u...

Embodiment 3

[0062] Example 3 Preparation of a 13-valent pneumococcal polysaccharide-protein conjugate vaccine

[0063]Single-carrier conjugates such as 4, 6B, 9V, 14, 18C, 19F, 23F were prepared according to known methods in the background art, wherein the carrier proteins were all CRM197. According to the method described in Example 1 or 2, prepare 1,3,5,6A, 7F,19A type polysaccharide double carrier conjugates, wherein 1,5,7F type polysaccharide adopts PspA as the second carrier, 3,6A,19A HiD was used as the second vector. The various conjugated products were formulated according to Table 3 below. After mixing these monovalent components, aluminum phosphate adjuvant was added at a final concentration of 0.5 mg / L.

[0064] Table 3 The carrier protein type of various antigens and the final content in the preparation

[0065]

[0066]

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Abstract

The invention provides a pneumococcal polysaccharide and protein conjugated vaccine and a preparation method thereof. The pneumococcus polysaccharide protein conjugated vaccine comprises one or more streptococcus pneumoniae capsular polysaccharide and protein coupled immune conjugates, and at least one of the immune conjugates is formed by coupling a single streptococcus pneumoniae capsular polysaccharide with two or more proteins. Compared with the existing pneumococcal conjugated vaccine, the pneumococcal polysaccharide and protein conjugated vaccine has stronger immunogenicity, and can cause immune response in a wider crowd; meanwhile, the two carrier proteins are covalently coupled through the polysaccharide, the protective protein antigen epitope induces higher immune response compared with the mixed injection of the two proteins, and through mutual synergy, the immunogenicity of the carrier protein is further improved and the immune response of the body to the polysaccharide is improved; the preparation method is simple, and meets the requirement of large-scale industrial production.

Description

technical field [0001] The invention relates to the field of immunology, in particular to a pneumococcal polysaccharide-protein conjugated vaccine and a preparation method thereof. Background technique [0002] Infections caused by pneumococci (pulmonary chain) are a major cause of morbidity and mortality worldwide. Pneumonia, febrile bacteremia, and meningitis are the most common manifestations of invasive pneumococcal disease, while bacterial spread within the respiratory tract can lead to middle ear infection, sinusitis, or recurrent bronchitis. Noninvasive manifestations are usually less severe but more common than invasive disease. The likelihood of pneumococcal disease flares during influenza is further increased by the spread of antibiotic-resistant infectious diseases and the fact that pneumococcal pneumonia often follows influenza infection. Diseases caused by Streptococcus pneumoniae have become an important public health problem worldwide. Pneumococcus has beco...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/385A61K39/116A61K39/09A61K47/48A61P31/04
CPCA61K39/092A61K47/6415A61K47/646A61P31/04
Inventor 朱涛宇学峰邱东旭毛慧华邵忠琦刘正
Owner CANSINO BIOLOGICS INC
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