A kind of bacterial polysaccharide conjugated vaccine and its preparation method and application

A technology of combining bacterial polysaccharides and vaccines, applied in the field of biomedicine, to achieve the effects of increasing the content of free polysaccharides, enhancing specific antibody titers, and good stability

Active Publication Date: 2022-03-15
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

[0008] At present, the carriers of conjugated vaccines have certain limitations, so it is necessary to study new, safe and effective vaccine carriers for the development of polysaccharide conjugated vaccines

Method used

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  • A kind of bacterial polysaccharide conjugated vaccine and its preparation method and application
  • A kind of bacterial polysaccharide conjugated vaccine and its preparation method and application
  • A kind of bacterial polysaccharide conjugated vaccine and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] 1. Bacterial polysaccharide crushing:

[0078] The original polysaccharide of meningitis capsular polysaccharide (CPS) has a large molecular weight and high viscosity, and it is easy to produce macromolecular cross-linking and reduce the stability. The invention adopts ultrasonic waves to crush polysaccharides into polysaccharides with uniform molecular weight. In the ultrasonic wave, the polysaccharide produces rapid mechanical movement, and the molecules are degraded into smaller molecules with the fluctuating high-speed vibration and shear force in the medium, and have no effect on the polysaccharide structural unit. The specific methods and steps are as follows:

[0079] (1) Weigh a certain amount of group C meningococcal capsular polysaccharide (CPS), and dilute it with 20mmol / L sodium phosphate buffer containing 0.15mol / L sodium chloride at pH=7.2, so that the final concentration of polysaccharide is 2mg / L mLl;

[0080] (2) Use No. 3 probe for ultrasonic crushi...

Embodiment 2

[0099] Characterization of Bacterial Polysaccharide Conjugate Vaccine

[0100] Molecular weight determination of polysaccharide-conjugated vaccines by multi-angle laser light scattering. The dn / dc value of HBc virus-like particles was 0.185mL / g, the dn / dc value of polysaccharide was 0.171mL / g, and the molecular weights of unbound HBc and CPS were 6.64×103kDa and 1.08×102kDa respectively by multi-angle laser scattering analysis. Compared with the unconjugated polysaccharide protein, the retention time of the three conjugated vaccines was significantly reduced, indicating that the polysaccharide protein conjugation was successful. In order to calculate the molecular weights of the three conjugated vaccines, the refractive index increments dn / dc of the three conjugated vaccines were calculated by the following formula I.

[0101]

[0102] R in the formula represents the ratio of polysaccharide to protein. The content of protein and polysaccharide in the conjugated vaccine wa...

Embodiment 3

[0104] Immune Effect Test of Bacterial Polysaccharides

[0105] Select 36 BALB / c mice, 18-22 g, female, and randomly divide them into 5 groups, 6 mice in each group. A total of 6 groups of experiments were carried out: pure sugar group (CPS group), CPS and HBc physical mixture group (CPS+HBc group), CPS-P2k-HBc group, CPS-P5k-HBc group, CPS-P10k-HBc group. All groups were injected subcutaneously, and each animal was injected with a vaccine containing 2.5 μg of polysaccharide, once a week, for a total of 3 injections. One week after the third injection, blood was collected by tail docking.

[0106] (1) First, test the immunogenicity of different groups of vaccines

[0107] Anti-capsular polysaccharide IgG, IgG1 and IgG2a antibody titers and polysaccharide-specific IgG2a / IgG1 antibody titer ratios in mouse serum were detected by ELISA. Test results such as Figure 3A , Figure 3B , Figure 3C and Figure 3D As shown, the antibody titer of the polysaccharide conjugate vacc...

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Abstract

The present invention provides a bacterial polysaccharide-conjugated vaccine and its preparation method and application. The bacterial polysaccharide-conjugated vaccine includes covalently bonded hepatitis B core antigen and bacterial polysaccharide; the bacterial polysaccharide-conjugated vaccine provided by the present invention can combine bacterial polysaccharide T cell-dependent antigens are converted to T cell-dependent antigens to generate Th1-type cellular immunity. Compared with common existing polysaccharide vaccines, the specific antibody titer and affinity are significantly enhanced (the affinity index measured by thiocyanate can reach 2mol / L, the highest can reach 2.17mol / L) and persistent effect, and has outstanding stability, stored at 37 ° C for 21 days, no significant increase in free polysaccharide content, easy to scale up production, with high practicality Value.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a bacterial polysaccharide-conjugated vaccine as well as a preparation method and application thereof. Background technique [0002] Meningococcus is the main cause of meningitis worldwide. Neisseria meningitidis invades the human body through the nasopharynx and forms purulent meningeal lesions in the meninges of the brain. It is the most common pathogenic bacteria of bacterial meningitis, and it is also the only bacteria that can cause large-scale epidemics of meningitis. After Neisseria meningitidis enters the blood circulation, meningococcus usually develops rapidly within 1-3 days, and is a disease with high morbidity and mortality. Even if antibiotic treatment is given after infection, 10% of patients will die within 24-48 hours of symptom onset; another 10%-20% of cured patients will have permanent sequelae such as neurological disorders or neurological deafness, etc., which will...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/385A61K39/095A61K39/116A61K39/39A61P31/04
CPCA61K39/095A61K39/385A61K39/39A61P31/04A61K2039/55516A61K2039/6031A61K2039/627A61K2039/70
Inventor 张松平苏志国杨延丽胥玲玲李正军
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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