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Preparation method of streptococcus pneumoniae capsular polysaccharide protein conjugate vaccine

A protein-conjugated vaccine, Streptococcus pneumoniae technology, applied in the direction of antibacterial drugs, bacterial antigen components, etc., can solve the problem of measurement error and other problems

Inactive Publication Date: 2014-06-04
TIANJIN TASLY PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The problem with these methods is that when the structure and properties of the measured sample are different from the molecular weight standard, it will bring obvious errors to the measurement results

Method used

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  • Preparation method of streptococcus pneumoniae capsular polysaccharide protein conjugate vaccine
  • Preparation method of streptococcus pneumoniae capsular polysaccharide protein conjugate vaccine
  • Preparation method of streptococcus pneumoniae capsular polysaccharide protein conjugate vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] 7F Sonication:

[0083] a Weigh 250 mg of 7F polysaccharide, add dipure water to prepare a solution with a concentration of 2.5 mg / ml, and dissolve overnight at 4°C. In an ice bath, the solution was treated with an ultrasonic wave with a power of 80-100 W and a frequency of 20 kHz for 23 minutes.

[0084] b freeze-dried

[0085]c Using SEC-RI-MALLS combined technology with 0.2M NaCl as the mobile phase and SRT-SEC 1000A as the chromatographic separation column, the ultrasonically degraded polysaccharides were detected at a flow rate of 0.5 mL min-1 and a column temperature of 30 °C . The detectors are multi-angle laser detectors (MALLS) and differential refractometers (RI).

[0086] d The results are as follows

[0087] Table 1 Quality comparison of type 7F polysaccharides before and after degradation

[0088]

[0089] It can be seen from Table 1 that the molecular weight of the 7F crude polysaccharide is greater than 2000KDa. After ultrasonic degradation, the m...

Embodiment 2

[0101] The method of Example 1 was used to prepare other 12 kinds of polysaccharide-protein conjugate vaccines except 7F.

[0102] The method is the same as in Example 1, but the polysaccharide is changed to one of 1, 3, 4, 5, 6A, 6B, 9V, 14, 18C, 19A, 19F or 23F

Embodiment 3

[0104] Quality control of the monovalent pneumonia polysaccharide protein conjugate obtained in Examples 1 and 2

[0105] The detection method of monovalent conjugate stock solution is as follows:

[0106] a molecular weight

[0107] Using SEC-RI-MALLS combined technology with 0.2M NaCl as the mobile phase and SRT-SEC 1000A as the chromatographic separation column, the molecular weight of the conjugate stock solution was detected at a flow rate of 0.5 mL min-1 and a column temperature of 30 °C . The loading volume is 100ul. The detectors are multi-angle laser scattering detectors (MALLS) and differential refractometers (RI).

[0108] b Determination of protein content

[0109] The BCA method was used to determine the protein content in the conjugate stock solution. A standard curve was drawn with BSA as the standard protein, and the protein content in the conjugate stock solution was calculated.

[0110] c Determination of polysaccharide content

[0111] The polysacchar...

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Abstract

The invention discloses a preparation method of a streptococcus pneumoniae capsular polysaccharide protein conjugate vaccine, which comprises the following steps: degrading extracted streptococcus pneumoniae capsular polysaccharide; and conjugating the degraded streptococcus pneumoniae capsular polysaccharide with a carrier protein, and preparing the vaccine preparation.

Description

Technical field: [0001] The invention relates to the preparation of a vaccine, in particular to a pneumonia polysaccharide protein conjugated vaccine and a preparation method thereof. Background technique: [0002] Streptococcus pneumoniae, which causes noninvasive infections such as sinusitis, otitis media, tracheitis, and pneumonia as well as invasive infections such as sepsis and meningitis, is a major cause of high morbidity and mortality in all age groups. pathogenic bacteria. In children's respiratory system, the incidence of pneumonia ranks second. World Health Organization (WHO) survey data show that among the 1.6 million people who die from Streptococcus pneumoniae infection every year, about 60% of them are children under 5 years old. It can be seen that Streptococcus pneumoniae can be called the "number one killer" that threatens children's health. At present, China is one of the five Asian countries with the highest incidence rate in the world. [0003] Accor...

Claims

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Application Information

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IPC IPC(8): A61K39/09A61P31/04
Inventor 赵秋敏孙倩李军强金永杰李亚冰刘贺军曹小丹李静李剑
Owner TIANJIN TASLY PHARMA CO LTD
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