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Beta-glucan modified meningitis polysaccharide conjugate vaccine and preparation method thereof

A technology combining vaccine and meningitis, applied in the field of biomedicine, can solve the problem of no patent application

Active Publication Date: 2015-04-29
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, there are no related literature reports and related patent applications on the chemical modification or physical mixing of immunomodulators and polysaccharide conjugate vaccines.

Method used

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  • Beta-glucan modified meningitis polysaccharide conjugate vaccine and preparation method thereof
  • Beta-glucan modified meningitis polysaccharide conjugate vaccine and preparation method thereof
  • Beta-glucan modified meningitis polysaccharide conjugate vaccine and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0019] Example 1: Preparation and separation and purification of β-glucan-modified meningitis polysaccharide protein-conjugated vaccine

[0020] (1) Preparation of meningitis polysaccharide-carrier protein conjugate vaccine (PS-TT)

[0021] Dissolve 5 mg of meningococcal capsular group Y polysaccharide (PS) in 1.25 ml of normal saline, adjust the pH of the solution to 10.8 with 0.5 mol / L of sodium hydroxide, add 10 microliters of 50% (w / v) cyanogen bromide, reacted for 30 minutes. During the activation process, as the pH decreased continuously, the pH value of the solution was maintained at 10.8 with 0.5 mol / liter of sodium hydroxide. After the activation, the pH value of the solution was adjusted to 8.5 with 0.5 mol / liter of hydrochloric acid, followed by adding 0.15 milliliters of adipic hydrazide solution with a concentration of 100 mg / ml, reacting overnight at room temperature ( figure 1 ). Subsequently, it was dialyzed three times for 12 hours in 20 mM phosphate buffe...

Embodiment 2

[0028] Example 2: Characterization of β-glucan modified polysaccharide conjugate vaccines

[0029] The purified product was identified with a Superose 6 gel filtration column (1.0 cm×30 cm), the eluent was 20 mM phosphate buffer (pH 7.4), and the flow rate was 0.5 ml / min. Such as figure 2 As shown, compared with the carrier protein TT, the peak time of PS-TT and PS-TT-G was significantly earlier. This indicates that the molecular weight of the carrier protein increases significantly after binding to the meningitis capsular polysaccharide.

[0030] use 1 H-NMR detection of polysaccharide conjugated vaccines. Group Y polysaccharides consist of →4-O-α-D-glucose p-(1→6)-β-D-sialic acid-2→repeating units, such as image 3 As shown in a, there are clear dextran terminal carbon proton peaks and sialic acid H3eq / ax resonance peaks at chemical shifts 3.3-4.2, and water peaks appear at 4.7ppm. At these positions, PS-TT and PS-TT-G also have the same peaks as PS, and at the same ti...

Embodiment 3

[0032] Example 3: Determination of immunogenicity of β-glucan modified polysaccharide conjugate vaccine

[0033] Take PS-TT, in which the concentration of PS is 10 μg / ml, both in a total volume of 3 ml, and physically mix with 30 μg and 90 μg of β-glucan, respectively. The mixed solutions were set as PS-TT / G1 group and PS-TT / G2 group respectively. Select 30 8-week-old female Blab / C mice weighing 15-22 g. They were randomly divided into 5 groups, namely PS group, PS-TT group, PS-TT-G group, PS-TT / G1 group and PS-TT / G2 group, with 6 mice in each group. Intraperitoneal injection, each injection containing 5 micrograms of polysaccharides, once a week, a total of 3 injections. After 21 days, blood was collected from the orbit. Anti-meningitis polysaccharide IgG, IgG1, IgG2a and IgM in mouse plasma were detected by ELISA.

[0034] (1) Determination of immunogenicity of PS-TT-G

[0035] Such as Figure 4 As shown in a, the IgG antibody titer produced by the first dose in the PS...

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Abstract

The invention relates to a beta-glucan modified meningitis polysaccharide conjugate vaccine and a preparation method thereof. The preparation method of the beta-glucan modified meningitis polysaccharide conjugate vaccine comprises the following steps: (1) activating meningococcus polysaccharide by cyanogen bromide, and then deriving by adopting adipic dihydrazide; (2) combining a derived meningococcus polysaccharide derivative with carrier protein; (3) activating beta-glucan; and (4) modifying polysaccharide-protein conjugate by the activated beta-glucan. By virtue of the steps, a novel and efficient meningitis polysaccharide conjugate vaccine can be prepared and can be used for preventing infection caused by epidemic cerebrospinal meningitis Neisseria gonorrhoeae.

Description

technical field [0001] The invention discloses a novel meningitis polysaccharide conjugate vaccine prepared based on the modification of β-glucan. The vaccine can be used to prevent epidemic meningitis and other diseases, and belongs to the field of biomedicine. Background technique [0002] Meningitis is an acute respiratory infectious disease caused by Neisseria meningitidis, which causes inflammation of the meninges. The onset cycle of epidemic meningitis is about 3-5 years, and a pandemic breaks out every 8-10 years. The condition of epidemic meningitis is complex and changeable, with varying degrees of severity. There are three types of clinical manifestations, namely common type, fulminant type, and chronic sepsis type. The incubation period is 1-7 days, generally 2-3 days. Neisseria meningitidis is hidden in the nasal and pharyngeal secretions of patients or carriers. It is mainly transmitted directly from the air by droplets through coughing, sneezing, talking, etc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/385A61K39/095A61K47/48A61P31/04
Inventor 胡涛季韶洋乔卫林
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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