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New hepatitis A inactivated vaccine virus strain and method for culturing same

A technology of hepatitis A virus and hepatitis A, applied in the direction of antiviral agent, inactivation/attenuation, and resistance to vector-borne diseases, etc., to achieve the effect suitable for large-scale industrial production and improve yield and quality

Active Publication Date: 2012-07-11
JIANGSU SIMCERE VAXTEC BIO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There are currently three manufacturers in my country producing live attenuated hepatitis A vaccine, and two manufacturers producing inactivated hepatitis A vaccine. There are differences in quality. my country has a large population, and the hepatitis A vaccine is still far from meeting the requirements of the domestic market.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The cultivation method of embodiment 1 hepatitis A virus strain JS-4 strain

[0020] The feces collected from clinical hepatitis A patients in the acute phase were prepared into 20% suspension, treated with 5% plant activated carbon, centrifuged at 10000rpm for 30min, the supernatant was absorbed and appropriate amount of antibiotics were added, and placed at 4°C for later use.

[0021] After the Vero cells grow into a single layer in the small square bottle, wash the cell surface once, inoculate each bottle with 2ml of feces suspension, absorb at 37°C for 2 hours, supplement the cell maintenance solution and culture at 36°C for 28 days to collect the virus, and change the medium every seven days during this period .

[0022] The samples separated and passaged in step (b) continue to be adapted for Vero cell culture. The difference is that each bottle of cells was inoculated with 1mL sample, adsorbed at 37°C for 2 hours, and cultured at 36°C for 28 days. After the 10th...

Embodiment 2

[0037] Example 2 The method for preparing hepatitis A inactivated vaccine virus antigen

[0038] Take out the cell cryopreservation tube from the Vero cell working seed bank, put it into 40°C water to dissolve quickly, then add it to the 199 culture medium pre-warmed at 37°C, plant it into the cell bottle, culture it at 37°C, and grow into single cells after 3-4 days After layering, amplify for several generations until the specified generation for production.

[0039] Add the above-mentioned cells to the hepatitis A virus strain JS-4 strain obtained in Example 1 at 0.2 MOI, inoculate them in a 3L three-layer flask after adsorbing the suspended cells at 37°C for 40 minutes, and culture them at 36°C, during which time, replace them every seven days. One time, 21 days to collect the poison, wherein the composition of the culture medium is 199 culture medium, containing 2-8% calf serum. After inactivation, the hepatitis A inactivated vaccine virus antigen is obtained.

[0040] ...

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Abstract

The invention provides a new hepatitis A virus JS-4 strain for preparing hepatitis A inactivated vaccine and a method for culturing the same. The strain is separated from deject a of a patient infected with acute hepatitis A and is transmitted to a Vero cell for adapted culture; through neutralization tests and other methods, the strain is proved to be hepatitis A virus; in the adaptation process, the subculture cycle of early generation is 28 days; when the strain is transmitted to the tenth generation, the cycle is shortened to 21 days; the strain is continuously cultured for ten generations; the culture temperature is between 35 and 36 DEG C; the cycle for virus propagation is shortened from 28 to 21 days; the antigen titer can reach 1:640-1:1,280; and the infectious titer is between 106.67 and 107.50 CCID50 / mL. The marmoset virulence test proves that the strain has weak virulence, is reliable in safety and has good immunogenicity and protective effect when the strain is used for producing the hepatitis A inactivated vaccine and is an ideal strain for producing the hepatitis A inactivated vaccine.

Description

technical field [0001] The invention relates to a novel hepatitis A virus JS-4 strain for preparing an inactivated hepatitis A vaccine and a cultivation method thereof. Background technique [0002] Hepatitis A is an acute infectious disease caused by hepatitis A virus infection that seriously endangers human health. It is especially an important public health problem in developing countries, manifested by high morbidity and localized epidemics, and even outbreaks causing death. Therefore, the prevention and treatment of hepatitis A occupies an important position in the global disease prevention and control. [0003] my country is a high prevalence area of ​​hepatitis A, at least 240,000 people suffer from hepatitis A every year, causing huge economic losses and social impact. Premier Wen Jiabao of the State Council proposed in the Government Work Report of the Fifth Session of the Tenth National People's Congress that infectious diseases such as hepatitis A and meningiti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/08A61K39/29A61P31/14
CPCY02A50/30
Inventor 张玉慧曹雨露周佳丽胡焱灵蒋建江王益民黄明
Owner JIANGSU SIMCERE VAXTEC BIO PHARMA
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