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Inactivated rabies antigen and preparation method thereof

A technology for inactivating antigens and rabies, applied in antiviral agents, pharmaceutical formulations, antibody medical ingredients, etc., can solve the problems of increasing production costs, difficult to popularize and apply, and high market sales prices, reducing production processes and reducing production costs. Effect

Active Publication Date: 2012-05-30
GUANGZHOU SOUTH CHINA BIOLOGICAL MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The imported veterinary inactivated vaccines currently registered in my country have better immune effects. The main preparation process is to use rabies virus (flury strain) or (ERA strain) as the vaccine strain, and then add inactivating agent after large-scale culture in rotary bottles. Inactivated, and then added aluminum glue, aluminum phosphate as an adjuvant or freeze-dried preparation; however, due to the limitation of the vaccine strain itself, the virus concentration of large-scale culture is not enough to meet the requirements of the inactivated vaccine to stimulate the body to produce sufficient protective antibodies Requirements, so 30-100 times of concentration is needed, which greatly increases the production cost and causes its market price to be higher, making it difficult to popularize and apply in my country, especially in rural areas.

Method used

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  • Inactivated rabies antigen and preparation method thereof
  • Inactivated rabies antigen and preparation method thereof
  • Inactivated rabies antigen and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0022] A method for preparing an inactivated rabies antigen, the preparation steps comprising:

[0023] (1) The titer of rabies virus dG strain virus liquid is adjusted to 5.0×10 with DMEM solution 6 FFU / mL, adjust the concentration of BHK-21 cell suspension to 5.0×10 with DMEM solution 5 individual / mL, the adjusted rabies virus dG strain virus solution is mixed with the adjusted BHK-21 cell suspension, wherein the adjusted virus solution volume accounts for 20% of the total volume of the mixed solution, and the virus and BHK-21 cell mixed solution 10% of the total volume was inoculated in a 10L spinner bottle, the speed of the spinner bottle was adjusted to 8 rpm, and cultured in an environment of 33°C. The virus titer after culture is not lower than 1.0×10 8 FFU / mL.

[0024] (2) After culturing for 72 hours, freeze and thaw the culture solution in the spinner flask three times at -50°C, centrifuge at 5000 rpm at 2°C for 30 minutes, remove the precipitated cell debris, col...

Embodiment 2

[0037] A method for preparing an inactivated rabies antigen, the preparation steps comprising:

[0038] (1) Adjust the titer of rabies virus dG strain to 5.0×10 with DMEM solution 6 FFU / mL, adjust the concentration of BHK-21 cell suspension to 5.0×10 with DMEM solution 5 cells / mL, the adjusted virus solution was mixed with the adjusted BHK-21 cell suspension, wherein the adjusted virus solution volume accounted for 10% of the total volume of the mixed solution, and the virus solution was mixed with the BHK-21 cell suspension. The mixed solution was respectively inoculated in the spinner bottle according to 10% of the total volume of the spinner bottle, and the speed of the spinner bottle was adjusted to 12 rpm, and cultured in an environment of 37°C. The virus titer of the cultured mixture was 1.0×10 8 -1.0×10 10 FFU / mL.

[0039] (2) After culturing for 96 hours, freeze-thaw the culture medium in the spinner flask five times at -20°C, centrifuge at 8,000 rpm at 4°C for 50 ...

Embodiment 3

[0051] A method for preparing an inactivated rabies antigen, the preparation steps comprising:

[0052] (1) Adjust the titer of rabies virus dG strain to 6.0×10 6 FFU / mL, adjust the concentration of BHK-21 cell suspension to 6.0×10 5 cells / mL, the adjusted virus solution was mixed with the adjusted BHK-21 cell suspension, wherein the adjusted virus solution volume accounted for 15% of the total volume of the mixed solution, and the virus and BHK-21 cell mixed solution was mixed according to the total volume. 10% was inoculated in a 10L spinner bottle, and the speed of the spinner bottle was adjusted to 9 rpm, and cultured in an environment of 37°C. The virus titer after culture should not be lower than 1.0×10 8 FFU / mL.

[0053] (2) After culturing for 90 hours, freeze and thaw the culture medium in the spinner flask 4 times at -20°C, centrifuge at 4°C at 6000 rpm for 40 minutes, remove the precipitated cell debris, collect the supernatant, and use the percentage by weight ...

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Abstract

The invention discloses an inactivated rabies antigen. A method for preparing the antigen comprises the following steps: mixing a rabies virus dG strain virus liquid with a titer of 5.0*10<6> to 1.0*10<7> FFU / mL and a BHK-21 cell suspension with a concentration of 5.0%*10<5> to 1.0*10<6> individuals / mL; cultivating for 72 to 96 h in the environment with a temperature of 33 to 37 DEG C; carrying out freeze thawing and centrifugation, and collecting a supernatant; regulating the pH of the supernatant to 7.4 to 7.6; adding beta-propiolactone with a final concentration of 0.025 to 0.05 V / V% drop by drop; and inactivating and hydrolyzing. According to the present invention, the method has the advantages of needing no condensation, simplifying production technology and saving production cost, and prepared inactivated vaccines can induce the generation of a high level of antibodies in a dog and can effectively protect the dog from virulent attack by the rabies virus. The inactivated rabies antigen prepared through the method allows condensation and purification to be omitted, production technology to be greatly simplified and production cost to be reduced, and is suitable for popularization and application in China.

Description

technical field [0001] The invention relates to an antigen for preventing canine rabies, in particular to an inactivated rabies antigen and a preparation method thereof. Background technique [0002] Rabies is a zoonotic disease caused by the rabies virus, with a fatality rate of almost 100%. The disease is prevalent in more than 100 countries and regions, and the number of deaths due to rabies in the world reaches 55,000 people every year. According to the latest figures released by the Ministry of Health of China, in 2008 and 2009, a total of 2,373 and 2,131 rabies deaths were reported nationwide, ranking among the top three in the number of reported deaths from 37 legal infectious diseases. About 90% of human rabies in our country is caused by dogs. cause. [0003] At present, there is no effective treatment for rabies, so timely vaccination is one of the most effective measures to prevent rabies. Starting from the source of virus infection, inoculating animals with ve...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/205A61P31/14
Inventor 薛素强郭霄峰梁昭平孙招金王斌李敏
Owner GUANGZHOU SOUTH CHINA BIOLOGICAL MEDICINE
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