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A combined inactivated vaccine of Zika virus and yellow fever virus

A technology of yellow fever virus and Zika virus, applied in antiviral agents, virus/bacteriophage, virus antigen components, etc., can solve the problem of no yellow fever virus and Zika virus joint inactivated vaccine development, side effects, etc., to achieve The effect of reducing the number of vaccination needles, low cost and wide coverage

Active Publication Date: 2021-03-19
SINOVAC BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Years of clinical application have shown that the attenuated yellow fever vaccine is safe and effective, but there are also serious side effects caused by the injection of the attenuated yellow fever vaccine, mainly yellow fever associated viscerotropic disease (yellow fever associated viscerotropic disease, YEL- AVD) and neurotropic disease (yellowfever associated neurotropic disease, YEL-AND)
There is no report on the development of a combined inactivated vaccine against yellow fever virus and Zika virus

Method used

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  • A combined inactivated vaccine of Zika virus and yellow fever virus
  • A combined inactivated vaccine of Zika virus and yellow fever virus
  • A combined inactivated vaccine of Zika virus and yellow fever virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Yellow fever virus 17D spot-picking and adaptive culture in Vero cells

[0028] In order to make the yellow fever virus 17D adapt to the growth on Vero cells better, the live attenuated vaccine of yellow fever virus was used to pick spot culture on Vero cells. The lyophilized live attenuated yellow fever virus 17D vaccine was restored to its original volume with water for injection. According to the virus titer of the known vaccine, the virus was serially diluted with serum-free 199 medium. Inoculate each dilution of the virus into a 6-well plate that has formed a monolayer of cells, inoculate 0.4ml per well, and inoculate at 37°C, 5% CO 2 Adsorb in the incubator for 1 hour, and then add serum-free carboxymethylcellulose working solution. After culturing for a period of time, select culture wells with less than 3 plaques under a microscope. The plaques were sucked up with a pipette and seeded on a monolayer of Vero cells for culture. When the cytopathic ef...

Embodiment 2

[0030] The titer and immunogenicity comparison of embodiment 2 yellow fever virus 17D in different passages on Vero cells

[0031] 1. Select the 1st, 4th, 7th, 10th, 13th, and 15th generations of adaptive passage on Vero cells to carry out virus titration test. Use the 199 cell maintenance liquid to serially dilute the virus, select 3 virus liquids with appropriate dilutions, and inoculate them into a 6-well plate in which the cells have formed a monolayer. Inoculate 2 wells for each dilution of virus, and set up 2 wells of cell control. At 37°C, 5% CO 2 Adsorb in the incubator for 1 hour, then add 4ml of carboxymethylcellulose solution to each well, and continue culturing in the incubator. After 6 days, take out the culture plate, discard the cover, add 1% crystal violet staining solution for staining, rinse with tap water, and dry in the air. PFU was calculated by counting plaques in wells with fewer than 30 plaques. The unit of virus titer is LgPFU / ml. Virus titer resu...

Embodiment 3

[0045] The cultivation optimization of embodiment 3 yellow fever virus 17D on Vero cells

[0046] For determining the optimal culture condition of yellow fever virus on Vero cell, inventor adopts JMP10 software to carry out experiment design according to inoculation mode, inoculation MOI, culture temperature and harvest time. The levels of each factor are as follows, the inoculation method is separate species and mixed species, the inoculation MOI is 1, 0.1, 0.01PFU / ml, the virus culture temperature is 33-37°C, and the harvest time is 25%, 50%, 75% of lesions, 100%. The optimal experimental conditions were determined by using the results of the virus titer of yellow fever virus. The experimental design and results are shown in Table 4:

[0047] Table 4 Yellow fever virus culture condition optimization result

[0048] temperature(°C) MOI(PFU / ml) Lesion degree virus titer 33 0.01 0.25 6.99 33 1 0.5 7.64 37 1 1 7.2 37 0.01 0.5 7.27 ...

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Abstract

The invention provides a Zika-virus-and-yellow-fever-virus combined inactivated vaccine, and belongs to the technical field of biological product preparing. 0.5 microgram / ml-10 microgram / ml of yellowfever viruses and 0.5 microgram / ml-10 microgram / ml of Zika viruses are contained in the combined inactivated vaccine. The invention also provides a preparing method for the Zika-virus-and-yellow-fever-virus combined inactivated vaccine; the preparing method includes the steps of Zika-virus and yellow-fever-virus inoculating, purifying and inactivating, wherein yellow-fever-virus inoculated MOI is0.01 PFU / ml to 1 PFU / ml, and Zika-virus inoculated MOI is 0.001 CCID<50> / ml to 0.1 CCID<50> / ml. By means of the Zika-virus-and-yellow-fever-virus combined inactivated vaccine, the Zika viruses and theyellow fever viruses can be immunized at the same time, infection and anaphylaxis which are caused by an attenuated vaccine are avoided, and the Zika-virus-and-yellow-fever-virus combined inactivatedvaccine has the good capacity for controlling diseases caused by the Zika viruses and the yellow fever viruses.

Description

technical field [0001] The invention belongs to the technical field of biological product preparation, in particular to a combined inactivated vaccine containing Zika virus and yellow fever virus. Background technique [0002] The yellow fever vaccines widely used in the world are prepared by inoculating chicken embryo tissue with 17D attenuated strain. my country has been producing yellow fever vaccines since 1953, and it has been more than 50 years. Years of clinical application have shown that the attenuated yellow fever vaccine is safe and effective, but there are also serious side effects caused by the injection of the attenuated yellow fever vaccine, mainly yellow fever associated viscerotropic disease (yellow fever associated viscerotropic disease, YEL- AVD) and neurotropic disease (yellowfever associated neurotropic disease, YEL-AND). YEL-AVD is an infection that causes acute outbreaks of the liver and other organs, resulting in a 60% mortality rate. And YEL-AND c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/12A61P31/12A61P31/14C12N7/00C12N7/06
CPCY02A50/30
Inventor 吴君兰吕哲王琳李育蓉郑翔姜德玉姚志东高强尹卫东
Owner SINOVAC BIOTECH
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