Epidemic encephalitis B and enterovirus 71 (EV71) combined vaccine

A combined vaccine, EV71 technology, applied in the field of preventive Japanese encephalitis EV71 combined vaccine and its preparation, can solve problems such as difficult to achieve 100% safety and increase compliance

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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] 4. Genetic engineering vaccines: the current full-virus vaccines contain viral genomes, and it is difficult to achieve 100% safety. Therefore, genetic engineering vaccines will attract people's attention
[0013] Combined vaccine can simplify the vaccination procedure, save manpower, material resources and reduce the suffering of immunized people, reduce the number of vaccinations and adverse reactions, JE EV71 combined vaccine has not been reported so far, so we have developed a new type of JE EV71 combined vaccine , in order to reduce the suffering of immunized persons (mainly infants) and increase their compliance

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Bioreactor perfusion culture and harvest of Japanese encephalitis virus

[0039] Take Vero cells that have grown to a monolayer cultured in a 15L flask or cell factory, trypsinize them into single cells, count the cells, and press 3 × 10 5 Cells / ml were inoculated into a bioreactor containing 5g / L Cytodex-1 microcarriers, and the culture medium was DMEM medium containing 10% newborn bovine serum, NaHCO 3 Adjust pH to 7.2, culture at 37°C, start 2 volumes / day perfusion culture after culturing for 2 days, the cell growth medium in the perfusion culture stage is DMEM medium containing 2% newborn calf serum, NaHCO 3 The pH was adjusted to 7.2, and the maintenance medium was changed on the 6th day. The maintenance medium was 199 medium containing 0.1% human serum albumin and NaHCO. 3 Adjust pH to 7.4. According to the optimized screening of the optimal inoculation concentration, culture conditions and harvest time of the virus, we selected the virus cell ratio to...

Embodiment 2

[0040] Example 2 Virus inactivation verification

[0041] 1) JE vaccine inactivation verification

[0042] Verification method: Inoculate 8 mice with 12-14 grams of the test product in the brain, each 0.03ml, and at the same time inoculate 0.5ml intraperitoneally, which is the first generation; 7 days later, 3 mice of the first generation were killed, and the brains were taken for After 7 days, 3 mice of the second generation were killed, and 6 mice were inoculated with 12-14 g per hour in the same method, which was the second generation. Three generations. After inoculation, observation was performed daily for 14 days, and those who died within 3 days were not counted (the number of dead animals should not exceed 20% of the total number of animals used in the experiment). Except for the sacrifice and non-specific death after inoculation, all mice in each generation survived as qualified.

[0043] 2) EV71 vaccine inactivation verification

[0044] Take the inactivated virus...

Embodiment 3

[0045] Example 3 titer determination

[0046] 1) Immunize Kunming mice with JE EV71 combined vaccine and reference vaccine according to the prescribed procedures, stimulate their immune response to produce antibodies, collect blood after expiration, separate serum, neutralize with Japanese encephalitis virus, and neutralize by plaque reduction Experiment to calculate the titer of the vaccine to be tested.

[0047] 2) SD rats were immunized with JE EV71 combined vaccine, immunized twice with an interval of 7 days, blood was collected on the 14th day after primary immunization, serum was separated, neutralized with EV71 virus, and the antibody titer was detected by microneutralization test.

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PUM

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Abstract

The invention relates to a combined vaccine prepared from epidemic encephalitis B viral vaccines and enterovirus 71 (EV71) viral vaccines and a preparation method thereof. An epidemic encephalitis B viral stock solution prepared from the epidemic encephalitis B virus through culture, harvest, inactivation, ultrafilter concentration and purification and an EV71 viral stock solution prepared from the EV71 virus through culture, harvest, inactivation, ultrafilter concentration and purification are prepared according to a certain proportion to form the combined vaccine. The combined vaccine can be used for simultaneously preventing and treating the hand-foot-and-mouth disease suffered by infants and children caused by the epidemic encephalitis B and the EV71 virus.

Description

Technical field: [0001] The invention relates to a combined vaccine and a preparation method thereof, in particular to a preventive JE EV71 combined vaccine and a preparation method thereof. Background technique: [0002] JE virus is mainly prevalent in Asia. About 3 billion people in the world live in endemic areas. China, India and Southeast Asia are all areas where JE virus is widely endemic. About 30 million newborns are born in these regions each year, with an estimated 50,000 infections and 10,000 deaths globally each year. At present, with the increase of the world's mobile population, the epidemic area of ​​Japanese encephalitis virus has expanded to Australia, and the incidence of tourists to the epidemic area has increased every year. Japanese encephalitis is a viral zoonotic disease transmitted by mosquito bites. The patient has acute onset, often involving the central nervous system, and the symptoms vary in severity, including latent infection, mild meningitis ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/295A61P31/14A61K39/12A61K39/125
Inventor 石慧颖
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