Thermostable vaccine with silica on surface and preparation method therefor

A silicon dioxide and heat-stabilized technology, which is applied to medical preparations with non-active ingredients, medical preparations containing active ingredients, and pharmaceutical formulas, can solve problems such as limiting effects and difficulty in storing vaccine liquids at room temperature, and achieve good Effects of immunogenicity and excellent thermal stability

Inactive Publication Date: 2016-01-27
ZHEJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the complexity of the calcium phosphate phase and the instability of its amorphous phase limit its effectiveness as a protective mineralization material, making it difficult to store vaccines at room temperature for a long time
Therefore, it is necessary to adopt more effective means to improve the thermal stability of the vaccine without damaging the biological activity of the vaccine. At present, it has not been possible to store the vaccine liquid at room temperature for more than one month and basically maintain the biological activity of the vaccine. technology

Method used

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  • Thermostable vaccine with silica on surface and preparation method therefor
  • Thermostable vaccine with silica on surface and preparation method therefor
  • Thermostable vaccine with silica on surface and preparation method therefor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Embodiment 1, preparation EV71 or OPV vaccine liquid

[0082]1. EV71 virus or OPV vaccine solution

[0083] 1. Infect RD cells or Vero (using DMEM culture fluid) with human hand-foot-mouth virus EV71 strain or poliovirus OPV vaccine strain, and cultivate in a cell culture box at 37° C. for 36-72 hours.

[0084] 2. The culture system obtained in step 1 is centrifuged (16000g, 10min), the supernatant is collected and filtered (using a filter membrane with a pore size of 0.22 microns), and the filtrate is collected, which is the virus vaccine liquid (after testing, the pH of the virus liquid is 7.2), it is named EV71 vaccine liquid or OPV vaccine liquid.

Embodiment 2

[0085] Example 2, using the EV71 virus liquid or OPV vaccine liquid obtained in Example 1 to prepare a silica mineralized vaccine liquid (that is, to prepare a thermally stable vaccine with silica on the surface through in situ silicon mineralization)

[0086] 1. Preparation of SilicifiedEV71 / OPV vaccine solution-A

[0087] Silicic acid can be prepared by acidification of sodium metasilicate or hydrolysis of alkyl silicate (TEOS or TMOS): dissolve sodium silicate with PBS or Tris buffer to a concentration of 30 mM, and adjust it to 8.0 by hydrochloric acid treatment; or The 1M silicate is prepared by shaking and hydrolyzing in 1mM hydrochloric acid for 15-30 minutes. In EV71 virus liquid or OPV vaccine liquid (the virus concentration is 1×10 8 PFU / mL, that is, 1×10 11 PFU / L) was added to the newly prepared silicic acid to make the concentration reach 2mM, and the pH was adjusted to about 5.5, which was the initial system; the initial system was incubated at room temperature ...

Embodiment 3

[0133] Embodiment 3, the biological characteristic experiment of SilicifiedOPV vaccine liquid

[0134] 1. Plaque characteristics

[0135] The SilicifiedOPV vaccine solution and the OPV vaccine solution were subjected to the following experiments respectively:

[0136] Diluted 10 times by volume (10 -1 、10 -2 、10 -3 、10 -4 , 10 -5 、10 -6 and 10 -7 ), and then seeded 300 μL / well on a monolayer of RD cells spread on a 12-well plate, placed at 37°C, 5% CO 2 Incubate in an incubator for 1-2 hours; then discard the virus solution, cover the cells with agar cover (i.e. low-sugar DMEM medium containing 1% agar and 2% FBS), place at 37°C, 5% CO 2 Incubated in an incubator for 3 days; then fixed with 4% formaldehyde solution at room temperature for 1 h, discarded the agar cover, and stained with crystal violet for 10 min at room temperature; observed plaque morphology and took pictures, and calculated plaque forming units (PFU); vaccine titer = (empty Plaque forming unit×diluti...

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Abstract

The invention discloses a thermostable vaccine with silica on a surface, a preparation method and applications, and especially relates to introduction of a series of uncontinuous amorphous silica with a nanostructure on a surface of a vaccine in a specific manner based on in-situ bionic silicon mineralization. The provided thermostable vaccine is characterized in that uncontinuous amorphous hydrated state silica with a nanostructure exists on the surface of the vaccine. Because amorphous silica can exist stably, a lot of surrounding water molecules can be tied on the surface of the surface through hydrogen bonds to form a protective hydration layer, and therefore a relatively stable microenvironment is provided for the vaccine inside. Experiments show that the silica mineral is effective at the aspect of vaccine protection, the room temperature storage time is prolonged by nearly 10 times, financial expense for vaccine refrigeration is lowered greatly. The preparation method has wide application prospects in development and preparation of a novel thermostable vaccine, and heat inactivation phenomena of vaccines can be solved efficiently in transportation and storage.

Description

technical field [0001] The present invention relates to biotechnology, in particular to a heat-stable vaccine with silicon dioxide on the surface and a preparation method thereof, in particular to a method of introducing discontinuous, amorphous hydrated silicon dioxide on the surface of the vaccine by in-situ mineralization A method of preparing a thermostable vaccine. Background technique [0002] Infectious diseases pose a serious threat to human life and cause more than 17,000,000 deaths per year, accounting for about 25% of all deaths in the world. Vaccination is one of the most effective medical interventions humans have developed to combat infectious diseases. Live attenuated vaccines (that is, attenuated strains of the virus) are the most important form of vaccine and played a huge role in the eradication of smallpox and polio. Because vaccines are temperature sensitive, cryopreservation is required. However, in impoverished areas that lack sufficient financial re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/04A61K47/42A61K47/32A61K39/00A61P31/12
CPCY02A50/30
Inventor 王广川秦成峰唐睿康李晓峰
Owner ZHEJIANG UNIV
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