Rotavirus polysaccharide-protein conjugate vaccine and preparation method thereof

A protein-conjugated vaccine and rotavirus technology, applied in the rotavirus polysaccharide-protein conjugate vaccine vaccine and its preparation, in the field of vaccines, can solve the problem that preventive polysaccharide and/or protein vaccines have not been reported and are not easy to enhance immunity , inability to induce immune memory effects, etc.

Active Publication Date: 2017-08-18
BRAVOVAX +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But this kind of polysaccharide vaccine has the following problems: (1) in young animals or infants, only weak immune response can be produced, or even no immune response, and the immune response will increase with age; (2) produce low-affinity antibodies (3) only produce short-lived immune response, do not have immune memory and immune enhancement effect during repeated vaccination; (4) easy to produce immune tolerance; (5) common adjuvant is not easy to play the role of immune enhancement to this antigen Effect, the 23-valent polysaccharide vaccine has a protection rate of 50-70% for invasive pulmonary chain infection, and can only be used for vaccination of people over 2 years old, while the peak age of onset of pneumonia is 6-12 months old; (6) has Polysaccharides with repeating structures are T cell-independent type 2 immunogens. Without the participation of T cells, they cannot induce immune memory effects, and the antibodies that stimulate the body are mainly IgM and IgG2, which cannot effectively activate the complement system
[0021] However, the above-mentioned traditional polysaccharide-protein binding technology has the following disadvantages: (1) The polysaccharide-AH derivative will continue to react with the polysaccharide activated by cyanogen bromide to form a self-polymer of the polysaccharide, which reduces the polysaccharide-protein binding efficiency; (2) EDAC mediates the combination of ADH-derived polysaccharides and carrier proteins, and at the same time, it is easy to cause self-crosslinking of polysaccharides and carrier proteins, thereby reducing the binding efficiency of polysaccharides and proteins; (3) polysaccharides and carrier proteins are large organisms The molecule is connected by ADH with only 6 carbon atoms in the middle. The structure of polysaccharides and proteins is bound to influence each other, making some important epitopes of polysaccharides easily shielded by proteins, thereby reducing the immunogenicity of polysaccharides
[0041] There are also reports that nano-microspheres can be used as carrier proteins and adjuvants for DNA vaccines, but there is no report on their application in prophylactic polysaccharide and / or protein vaccines

Method used

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  • Rotavirus polysaccharide-protein conjugate vaccine and preparation method thereof
  • Rotavirus polysaccharide-protein conjugate vaccine and preparation method thereof
  • Rotavirus polysaccharide-protein conjugate vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0109] 1. Preparation of Magnetic Nanospheres

[0110] 1. Take 2.24g FeSO 4 -7H 2 O and 3.24 g FeCl 3 -6H 2 O dissolved in 10 mL and 15 mL of dddH 2 In O, mix well after dissolving, add 100mL of dddH filtered to remove oxygen 2 O;

[0111] 2. In N 2 Stir for 5 minutes under the protection of the solution, add 50mL1mol / L NaOH solution at one time, adjust the pH of the solution to 9-10, increase the stirring speed to 200-250r / min, and continue stirring for 30min;

[0112] 3. Transfer the reaction vessel to a 65-70°C water bath and continue to 2 Stirring and aging under the protection of 30min;

[0113] 4. After the reaction, set the volume to 100mL, and observe the synthesis of magnetic particles under a microscope;

[0114] 5. Dissolve 400mgPLGA in 10mLEA solvent as the oil phase (O), add 3mL of the above magnetic particle solution as the inner water phase (W 1 ), using an ultrasonic cell breaker (120W, 60s) to carry out colostrum in an ice-water bath to prepare colos...

Embodiment 2

[0158] The difference between this example and Example 1 is that the carrier protein of the rotavirus polysaccharide-protein conjugate vaccine is rotavirus carrier protein and tetanus toxoid carrier protein.

[0159] The composition analysis results of the prepared rotavirus polysaccharide-protein conjugate vaccine are consistent with the results obtained in Example 1 within the theoretical error range.

Embodiment 3

[0161] The difference between this example and Example 1 is that the carrier protein of the rotavirus polysaccharide-protein conjugate vaccine is rotavirus carrier protein and Haemophilus influenzae surface protein HiD.

[0162] The composition analysis results of the prepared rotavirus polysaccharide-protein conjugate vaccine are consistent with the results obtained in Example 1 within the theoretical error range.

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Abstract

The invention discloses rotavirus polysaccharide-protein conjugate vaccine that is a conjugated vaccine formed by connecting multivalent pneumococcus polysaccharide and two or more carrier proteins via connectors, wherein the connectors are magnetic nano microspheres; one carrier protein is rotavirus protein. A preparation method of the rotavirus polysaccharide-protein conjugate vaccine comprises: coupling the multivalent pneumococcus polysaccharide and the two or more carrier proteins (one of which is rotavirus carrier protein) respectively to the magnetic nano microspheres. The preparation method of the rotavirus polysaccharide-protein conjugate vaccine is simple; the rotavirus polysaccharide-protein conjugate vaccine using the nano microspheres as the connectors can enhance Th1-type immune response in mice and immune persistence, specificity and affinity of polysaccharide specific antibodies; in addition, the vaccine may also induce mice to produce rotavirus antibodies; the vaccine has the preventive effects of two vaccines; accordingly the vaccine has a promising application prospect.

Description

technical field [0001] The invention relates to a vaccine, in particular to a rotavirus polysaccharide-protein conjugated vaccine and a preparation method thereof, belonging to the field of biotechnology. Background technique [0002] 1. The harm of microorganisms to the human body and countermeasures [0003] Microorganisms usually refer to those biological groups whose individual volume diameter is generally less than 1mm. They have a simple structure, most of which are single cells, and some do not even have a cell structure. To clarify their morphology and structure; Among them, pathogenic microorganisms refer to pathogenic microorganisms that can cause diseases of humans, animals and plants. The pathogenicity of a pathogen depends on its ability to invade the host and reproduce in vivo and resist host resistance without being eliminated by it. The pathogenicity of microorganisms has species characteristics, and the degree of pathogenicity is called virulence. The est...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/116A61K39/09A61K39/385A61K47/64A61K47/69A61P31/04A61K39/295A61K39/15A61P31/14
CPCA61K39/09A61K39/12A61K39/385A61K2039/57A61K2039/575A61K2039/6037A61K2039/6068A61K2039/6075A61K2039/627A61K2039/70C12N2720/12334C12N2720/12341
Inventor 刘昊智王文灏程超吴克
Owner BRAVOVAX
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