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Thermally-stable emulsion of antigen as well as preparation method and application of thermally-stable emulsion

A heat-stable, antigen-based technology, applied in emulsion delivery, pharmaceutical formulations, antibody medical components, etc., can solve the problems of long antigen release time, expensive equipment, complicated operation steps, etc., to achieve good safety and stability, reduce Transport and storage costs, effects of good antigen thermal stability

Active Publication Date: 2019-02-22
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, aluminum salt adjuvants also have application limitations: 1) The antigens carried in aluminum salt adjuvants are taken up by antigen-presenting cells (APCs) in the form of soluble antigens, which mainly enhance the humoral immune response, but cannot induce cellular immune responses. immune response
2) The slow-release time of the antigen in the aluminum salt adjuvant is longer, resulting in unsatisfactory antigen presentation and delayed immune response
At present, the existing methods for preparing heat-stable vaccines are mainly obtained by preparing dry powder vaccines, which generally require complex operation steps and expensive instruments to achieve
And many vaccines, especially those mixed with adjuvants, cannot be preserved in the form of dry powder
[0013] In the current prior art, there is no research on the formation of vaccine adjuvants based on biomimetic mineralization of vaccines combined with Pickering emulsions, and the known Pickering emulsion systems have not been designed and optimized according to the thermal stability requirements of vaccine preparations

Method used

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  • Thermally-stable emulsion of antigen as well as preparation method and application of thermally-stable emulsion
  • Thermally-stable emulsion of antigen as well as preparation method and application of thermally-stable emulsion
  • Thermally-stable emulsion of antigen as well as preparation method and application of thermally-stable emulsion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] In this example, the biomimetic mineralization of aluminum hydroxide particles was used to prepare the heat-stable emulsion of antigen

[0093] (1) Preparation of aluminum hydroxide particles by alkoxide hydrolysis-hydrothermal method: weigh a certain amount of deionized water with a measuring cylinder and place it in a 100mL three-neck flask, heat to 85°C, and then add aluminum isopropoxide powder. Aluminum isopropoxide and deionized water were mixed at a molar ratio of 1:100, stirred for 2 hours, and cooled to room temperature naturally. The above reaction solution is put into a stainless steel high-pressure container lined with polytetrafluoroethylene, and subjected to hydrothermal treatment at a certain temperature. After the hydrothermal reaction is completed, the material is naturally cooled, centrifuged to separate the solid phase product, washed with deionized water three times, and vacuum-dried to obtain aluminum hydroxide particles. Aluminum hydroxide particl...

Embodiment 2

[0101] In this example, the biomimetic mineralization of calcium phosphate particles was used to prepare an antigen heat-stable emulsion

[0102] (1) Preparation of hollow calcium phosphate particles by template method:

[0103] After mixing 3.0g Tween 80, 0.25g PEG 6000, 3.0mL 0.5mol / L Tris-HCl (pH8.0) and 1.5mL deionized water and stirring evenly, ultrasonic 20min to form non-ionic surfactant vesicles, drop into 3.55mL CaCl 2 (0.175mol / L) solution, after stirring for 0.5h, add 3.55mL Na 2 HPO 4 (0.175mol / L) solution can obtain the suspension of sodium phosphate particle, add 0.83mL MgCl 2 solution (0.075mmol / L) to stabilize the newly formed calcium phosphate particles, continue to stir for 2h, centrifuge and wash, and vacuum dry to obtain hollow calcium phosphate particles. The average particle size of the particles is 210nm, the shell thickness is 30-40nm, and the PDI value of the particles is 0.289. The prepared particles are hollow spheres.

[0104] (2) Biomimetic m...

Embodiment 3

[0109] In this example, peanut-like calcium carbonate particles were used to prepare the heat-stable antigen emulsion by biomimetic mineralization

[0110] (1) Prepare peanut-shaped calcium carbonate particles by liquid phase direct mixed precipitation method:

[0111] Weigh calcium acetate and trisodium citrate with an electronic balance and dissolve them in 200mL of distilled water (the mass concentration of trisodium citrate is 10wt.% and 30wt.% respectively), and add 10wt.% sodium carbonate aqueous solution (50mL ), stirred (300rpm) for 3h, filtered, washed the precipitate three times with distilled water and absolute ethanol, and dried at 70°C to obtain peanut-shaped calcium carbonate particles. The length of the particles is 7.2 μm, the ratio of the long axis to the short axis is 2:1, and the morphology of the prepared particles is peanut-like.

[0112] (2) Biomimetic mineralization of proteins:

[0113] Accurately weigh 3.1 g of calcium carbonate particles with an ele...

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Abstract

The invention provides a thermally-stable emulsion of an antigen as well as a preparation method and an application of the thermally-stable emulsion. The thermally-stable emulsion of the antigen comprises a water phase, an oil phase and inorganic particles combined with the antigen; the oil phase is wrapped by the water phase; the inorganic particles combined with the antigen are dispersed on an interface of the water phase and the oil phase; and the antigen and the inorganic particles are combined in an electrostatic adsorption way. By using the emulsion provided by the invention, the thermalstability of the antigen of an organism can be remarkably improved; structural water molecules in a mineral phase are tightly connected to the peripheries of mineralized proteins, so that the hydrogen bond exchange rate of protein molecules and water molecules in an external environment is reduced, the destruction of a thermal motion process to the structures of the proteins is reduced, the stability of a vaccine in a high-temperature environment is improved, the valid period of vaccine storage is prolonged, and the vaccine can be stored at 25 DEG C for three weeks or longer; in addition, theemulsion can also be applied as a vaccine adjuvant, a drug delivery carrier or a controlled-release carrier, a surfactant is prevented from being used, and the poisonousness to human bodies and environment pollution can be reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an antigen heat-stable emulsion and a preparation method and application thereof. Background technique [0002] Infectious diseases have long been one of the major threats to human health. As one of the effective means to deal with infectious diseases, vaccination plays an extremely important role in the prevention and control of diseases. However, most of the early vaccines were attenuated or inactivated whole virus vaccines. Although they can stimulate strong protective antibody levels and cellular immune responses, there are also risks of incomplete virulence recovery and inactivation. Therefore, with the development of biotechnology, split vaccines, subunit vaccines, recombinant protein vaccines, synthetic peptide vaccines, and polysaccharide conjugate vaccines with higher safety have gradually replaced attenuated and inactivated vaccines in clinical practice. Especially in recen...

Claims

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Application Information

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IPC IPC(8): A61K39/39A61K47/02A61K9/107A61P37/06A61P37/04
CPCA61K39/39A61K47/02A61P37/04A61P37/06A61K9/107A61K2039/55505Y02A50/30
Inventor 马光辉吴颉吴楠苗春宇周炜清
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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