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Meningitis polysaccharide conjugate vaccine and preparing method thereof

A combined vaccine and meningitis technology, applied in the field of biomedicine, can solve problems such as restricting the development of polysaccharide conjugate vaccines, reducing polysaccharide-protein binding efficiency, and reducing polysaccharide immunogenicity.

Inactive Publication Date: 2014-04-02
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned polysaccharide-protein combination technology has the following disadvantages: (1) EDAC mediates the binding of ADH-derived polysaccharides and carrier proteins, and at the same time easily causes the self-crosslinking of the carrier protein to form polymers of the carrier protein, thereby Reduce the polysaccharide-protein binding efficiency; (2) The polysaccharide and the carrier protein are both macromolecules, and the middle is connected by ADH with only 6 carbon atoms in length. The carrier protein will inevitably shield some important epitopes of the polysaccharide, thereby reducing the polysaccharide. Immunogenicity
These deficiencies limit the further development of polysaccharide conjugate vaccines

Method used

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  • Meningitis polysaccharide conjugate vaccine and preparing method thereof
  • Meningitis polysaccharide conjugate vaccine and preparing method thereof
  • Meningitis polysaccharide conjugate vaccine and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Preparation and separation and purification of group A meningitis polysaccharide conjugate vaccine

[0022] (1) Activation and derivation of meningitis group A polysaccharides

[0023] The preparation reaction of group A meningitis polysaccharide conjugate vaccine is as follows: figure 1 shown. Dissolve 16 mg of group A meningococcal capsular polysaccharide in 4 ml of normal saline, and adjust the pH to 10.8 with 0.2 M NaOH. Add 32 microliters of 50% (w / v) cyanogen bromide solution for activation, and react at room temperature for 1 hour. During the activation process, the pH of the solution was maintained at 10.8 with 0.2M NaOH as the pH decreased continuously. After the activation, the pH value of the solution was adjusted to 8.5 with 0.5 M hydrochloric acid, and 1 ml of ethylenediamine solution with a concentration of 32 mg / ml was added. The pH value of the solution was reduced to about 5.0, and then the pH value of the solution was adjusted to 8.0 wit...

Embodiment 2

[0031] Example 2: Preparation and separation and purification of group C meningitis polysaccharide conjugate vaccine

[0032] (1) Activation and derivation of group C meningitis polysaccharides

[0033] The preparation reaction of group C meningitis polysaccharide conjugate vaccine is as follows: figure 1 shown. Dissolve 16 mg of group C meningococcal capsular polysaccharide in 4 ml of normal saline, and adjust the pH to 10.8 with 0.2 M NaOH. Add 32 microliters of 50% (w / v) cyanogen bromide solution for activation, and react at room temperature for 1 hour. During the activation process, the pH of the solution was maintained at 10.8 with 0.2M NaOH as the pH decreased continuously. After the activation, the pH value of the solution was adjusted to 8.5 with 0.5 M hydrochloric acid, and 1 ml of ethylenediamine solution with a concentration of 48 mg / ml was added. The pH value of the solution was reduced to about 5.0, and then the pH value of the solution was adjusted to 8.0 wit...

Embodiment 3

[0041] Embodiment 3: SD-PAGE electrophoresis identification polysaccharide conjugated vaccine

[0042] Group A and C polysaccharide conjugate vaccines were identified by SDS-PAGE electrophoresis. like Figure 4 As shown, tetanus toxoid showed two main electrophoresis bands (lane 2) on the SDS-PAGE electrophoresis graph. Group A polysaccharide conjugate vaccine (lane 3) and group C polysaccharide conjugate vaccine (lane 4) also showed 2 major electrophoresis bands, but their migration rates were significantly slower than those corresponding to tetanus toxoid. This indicates that the group A and group C polysaccharide conjugate vaccines have very high molecular weight and are significantly larger than tetanus toxoid.

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Abstract

The invention describes a method for preparing a meningitis polysaccharide conjugate vaccine. The meningitis polysaccharide conjugate vaccine is developed based on the method. The method comprises the following steps: (1) activating meningococcus polysaccharides by cyanogen bromide, then deriving the meningococcus polysaccharides which are activated by the cyanogen bromide by ethanediamine, finally deriving the polysaccharides by a reagent which is of a structure of succinimidyl ester-R-maleimide; (2), sulfhydrylating carrier proteins; (3) combining the derived meningococcus polysaccharides with the sulfhydrylated carrier proteins. As conjugation bridges between the meningococcus polysaccharides and the carrier proteins are very long, the spatial shielding effect of the carrier proteins on the antigenic epitopes of the polysaccharides is reduced, and the original immunization property of the polysaccharide conjugate vaccine is improved.

Description

technical field [0001] The invention provides a combination technology for preparing epidemic meningitis capsular polysaccharide conjugate vaccine; based on the combination technology, a meningitis capsular polysaccharide conjugate vaccine with high immunogenicity is prepared, which can be used for meningitis and the like The immune prevention of important diseases belongs to the field of biomedicine. Background technique [0002] Meningitis is an acute respiratory infectious disease caused by Neisseria meningitidis. Neisseria meningitidis is mainly transmitted by coughing, sneezing or kissing, invades the blood circulation from the nasopharynx, and finally is confined to the meninges and spinal cord membranes, forming purulent cerebrospinal meningopathy. Common symptoms include high fever, severe headache, and stiffness in the back of the neck. Drowsiness, vomiting, photophobia, or skin rash may also occur. This disease can cause brain damage or even death, and it is foun...

Claims

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Application Information

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IPC IPC(8): A61K39/385A61K39/095A61K47/48A61P31/04
Inventor 胡涛季韶洋苏志国
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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