Zika virus and encephalitis B virus combined inactivated vaccine

A technology for Japanese encephalitis virus and Zika virus, applied in the field of combined inactivated vaccine, can solve the problem of no joint inactivated vaccine development of Japanese encephalitis virus and Zika virus, so as to reduce the number of vaccinations, cause good disease, prevent and control disease-causing effects

Inactive Publication Date: 2018-06-22
SINOVAC RES & DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

JE virus and Zika virus belong to the yellow fever virus genus, and both are togaviruses. In my country, JE virus vaccine is one...

Method used

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  • Zika virus and encephalitis B virus combined inactivated vaccine
  • Zika virus and encephalitis B virus combined inactivated vaccine
  • Zika virus and encephalitis B virus combined inactivated vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The cultivation technique of embodiment 1 Japanese encephalitis virus vaccine

[0030] After the Vero cells are cultured to a single layer on the microcarrier, the cell culture medium is discarded, the cell surface is washed with PBS, and the JE virus is inoculated, and the second-generation working virus of the JE rat brain virus strain P3 is used to pass on the Vero cells. species, the inoculation MOI is 0.01CCID 50 / ml, the pH of the virus culture medium was 7.6, and it was cultured at 35°C. The supernatant was harvested for the first time 2 days after inoculation with JE virus, and the harvested volume was 80% of the total liquid volume. Afterwards, the supernatant was harvested every two days according to the aforementioned instructions, for a total of 5 harvests. The harvested supernatants of each batch were combined to obtain the Japanese encephalitis virus harvesting liquid.

Embodiment 2

[0031] The purification of embodiment 2 Japanese encephalitis virus

[0032] After the JE virus harvest solution was filtered through a deep membrane stack to remove cell debris and large clumps, the cell supernatant was harvested. Concentrate by ultrafiltration with a molecular weight cut-off of 300,000 to obtain a concentrate. After gel filtration chromatography and ion exchange chromatography.

[0033] Among them, gel filtration chromatography can use GE Sepharose 4Fast Flow, Sepharose 4FastFlow, Sephacryl S-300HR, Sephacryl S-400HR, Sephacryl S-500HR and other high-throughput and high-capacity fillers with suitable separation range and easy scale-up. The optional fillers for ion exchange chromatography are GE DEAE, CM and ANX fillers. The specific operation is as follows.

[0034] (1) At first the Japanese encephalitis virus vaccine concentrate is filtered through the chromatographic column, the ultraviolet detection wavelength is 280nm, the flow velocity is 6cm / h, and ...

Embodiment 3

[0042] The inactivation process of embodiment 3 Japanese encephalitis virus

[0043] The Japanese encephalitis virus purification solution prepared in Example 2 was inactivated, and inactivated according to two inactivation methods, respectively, β propiolactone with a final concentration of 0.025% was inactivated at 4°C and the final concentration was 30-180ug / ml formaldehyde 25 Inactivation at ±3°C, samples were taken at 0, 6, 12, 24, 36, 48, 72, and 96 hours, and the virus titer was detected. The results were analyzed using SPSS17.0 software, expressed as (x±s), according to Analysis of variance and t-test were used, and P<0.05 was considered statistically significant. The results are shown in Table 2. After 6 hours of formaldehyde inactivation, the virus titer began to decline. After 24 hours, most of the viruses had been inactivated, and no virus titer was detected after 48 hours. No virus can be detected after 12 hours of β-propiolactone inactivation.

[0044] Samples ...

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Abstract

The invention provides a zika virus and encephalitis B virus combined inactivated vaccine, and belongs to the technical field of preparation of biological products. The combined vaccine consists of 0.5-10ug/ml of encephalitis B virus and 0.5-10ug/ml of zika virus. The invention also provides a preparation method of the zika virus and encephalitis B virus combined inactivated vaccine; the preparation method comprises steps of inoculating, purifying and inactivating the encephalitis B virus and the zika virus, wherein inoculated MOI of the encephalitis B virus is 0.001-0.1CCID50/ml; and inoculated MOI of the zika virus is 0.001-0.1CCID50/ml. The vaccine provided by the invention can be used for simultaneously immunizing the zika virus and the encephalitis B virus, so that immunizing times are reduced and infection and allergic reactions caused by attenuated vaccines are avoided; and the combined inactivated vaccine has a good capacity of preventing and controlling diseases caused by thezika virus and the encephalitis B virus.

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 Japanese encephalitis virus. Background technique [0002] Zika virus has been discovered since the middle of the 20th century. It is mainly prevalent in tropical regions such as South America, Africa, and Southeast Asia, and there is no vaccine for protection. From 2015 to 2016, there were thousands of cases of neonatal microcephaly caused by Zika virus in Brazil, causing great panic. With the improvement of my country's economic level, people have more and more opportunities to travel to the aforementioned areas for business and travel, and the chances of being infected with Zika virus will increase. At present, there is no Zika vaccine on the market, and there is an urgent need to develop a Zika vaccine that can protect people of all ages and provide security for the health of our people. [0003...

Claims

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Application Information

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IPC IPC(8): A61K39/12A61K39/39A61P31/14C12N7/00C12N7/02C12N7/06
CPCA61K39/12A61K39/39A61K2039/5252A61K2039/55505A61K2039/70C12N7/00C12N2770/24134C12N2770/24151C12N2770/24163C12N2770/36034C12N2770/36051C12N2770/36063A61K2300/00Y02A50/30
Inventor 姜德玉吴君兰吕哲马铭江单慈恩王琳高强尹卫东
Owner SINOVAC RES & DEV
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