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A kind of chitosan oligosaccharide vaccine adjuvant based on chemical coupling and its application

A technology of vaccine adjuvant and chitosan oligosaccharide is applied in the field of biomedicine to achieve the effects of mature and stable preparation method, good adjuvant effect and enhanced immune response

Active Publication Date: 2020-01-10
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

Although simple physical mixing can exert immune adjuvant activity, it is not necessarily the best way to exert its adjuvant activity

Method used

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  • A kind of chitosan oligosaccharide vaccine adjuvant based on chemical coupling and its application
  • A kind of chitosan oligosaccharide vaccine adjuvant based on chemical coupling and its application
  • A kind of chitosan oligosaccharide vaccine adjuvant based on chemical coupling and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: the preparation of chitosan oligosaccharide vaccine adjuvant

[0030] 1. Preparation of chitosan oligosaccharide adjuvant:

[0031] (1) Thiolylation derivatization of oligochitosan COS

[0032]Weigh the required mass of COS (mass ratio COS:PCV=1:1) and 2-iminosulfane hydrochloride (IT) respectively, take IT by the molar ratio of IT:COS=5:1, and use 20mM PBS (pH 7.4) was dissolved and mixed, and reacted at room temperature for 3 hours.

[0033] After the reaction time is up, use a G25 desalting column to elute to remove unreacted small molecules.

[0034] (2) Maleimidization Derivation of Porcine Circular Inactivated Vaccine PCV

[0035] Draw the PCV of 3mL1.7mg / mL according to the required amount, weigh 3-benzoylmaleimide-N-hydroxysuccinimide (MBS) by the molar ratio of MBS:PCV=100:1, First dissolve in DMF and then dissolve with 20mM PBS (pH 7.4), add PCV to mix, and react at 4°C for 3 hours, wherein the reaction buffer is 20mM PB (pH 7.2), and the vol...

Embodiment 2

[0041] Embodiment 2: the vaccine adjuvant activity determination of chitosan oligosaccharide vaccine adjuvant

[0042] The immune activity of the chitosan oligosaccharide conjugate in Example 1 was detected, the mice were immunized by intramuscular injection, and the antibody titer was determined. The specific method is as follows:

[0043] Experimental animals: C57BL / 6 mice, 4-6 weeks old, 6 mice / group, female.

[0044] Control solvent: phosphate buffer saline (PBS).

[0045] Dosage: porcine circular inactivated vaccine (PCV): 100 μg / mouse (mouse); chitooligosaccharide: 100 μg / mouse; chitooligosaccharide conjugate 1 (PCV-COS-1): 100 μg / mouse; shell Oligosaccharide conjugate 2 (PCV-COS-2): 100 μg / mouse; oil emulsion adjuvant (ISA206): 100 μg / mouse.

[0046] Grouping: (1) Blank control group (PBS); (2) Individual vaccine group (PCV): PBS+PCV; (3) Physical mixture group (PCV / COS): COS+PCV; (4) PCV-COS-1 Group: PBS+PCV-COS-1; (5) PCV-COS-2 group: PBS+PCV-COS-2; (6) oil emulsi...

Embodiment 3

[0060] Embodiment 3: Pathological section research of chitosan oligosaccharide vaccine adjuvant

[0061] The injection sites of each group of mice in Example 2 were collected, and then treated with conventional pathological methods. Specifically, the tissue at the injection site was fixed with 10% formalin solution, embedded in paraffin and cut into 4-mm sections, then stained with HE and observed and photographed with a microscope (Leica) under bright field.

[0062] Experimental results:

[0063] In order to verify whether there is possible adverse reaction in the chitosan oligosaccharide adjuvant of the present invention, collect the injection site of each immunization group mouse and carry out the comparison of pathological section ( Figure 5 ). The results showed that the oil-emulsion adjuvant group had an obvious inflammatory reaction at the injection site, but there was no significant change in the chitosan-oligosaccharide conjugate group and other groups. It shows ...

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Abstract

The invention belongs to the technical field of biological medicines and particularly relates to a chitosan oligosaccharide vaccine adjuvant based on chemical coupling and an application of the chitosan oligosaccharide vaccine adjuvant. The chitosan oligosaccharide vaccine adjuvant comprises sulfhydrylated chitosan oligosaccharide and a heteromorphic bifunctional chelating agent, and sulfhydrylated chitosan oligosaccharide is coupled with a vaccine through the heteromorphic bifunctional chelating agent. The chitosan oligosaccharide vaccine adjuvant can enhance immune response, has a good adjuvant function and can be used as an adjuvant for an attenuated vaccine, a protein vaccine, a nucleic acid vaccine or a polypeptide vaccine. Chitosan oligosaccharide in the chitosan oligosaccharide vaccine adjuvant can be produced on a large scale, and a preparation method is manure and stable.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular, the invention relates to a chemical coupling-based chitosan oligosaccharide vaccine adjuvant and application thereof. Background technique [0002] Porcine circovirus type 2 (PCV2) is the main pathogen that causes multisystem wasting syndrome in weaned piglets, and it is often co-infected clinically with porcine reproductive and respiratory syndrome virus, porcine foot-and-mouth disease virus and porcine parvovirus. At the same time, PCV2 is also closely related to diseases such as porcine dermatitis and nephrotic syndrome, porcine respiratory disease syndrome, sow reproductive disorder, congenital tremor in piglets, necrotizing lymphadenitis, granulomatous enteritis and exudative epidermatitis. PCV2-related diseases are collectively referred to as porcine circovirus disease (Expert review of vaccines, 2015, 14:473-487). The disease was first discovered in my country in 2000. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/39A61K39/385A61K39/12A61P31/20A61P37/04
CPCA61K39/12A61K39/385A61K39/39A61K2039/55511C12N2750/10034
Inventor 杜昱光张贵强胡涛贾培媛刘洪涛
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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