Method for preparing enterovirus vaccines

An enterovirus and vaccine technology, applied in the field of enterovirus vaccine preparation, can solve the problems of yield limitation, low efficiency, low reproduction titer and the like, and achieve the effects of saving consumption, simple production process and convenient purification

Active Publication Date: 2014-08-13
ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the former, the reproduction titer of EV71 virus in this diploid cell is low, the yield is limited, and it is difficult to expand production; for the latter, since Vero cells are passaged cells, the DNA content of passaged cells contained in the vaccine needs to be below 30ng. However, it is difficult to remove the DNA in the virus fluid, resulting in higher production costs and lower efficiency

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: Preparation of EV71 monovalent vaccine

[0022] (1) Preparation of cell monolayer

[0023] Take gerbils aged 10-15 days, cut off the carotid artery and bleed to death, soak in 0.2% (v / v) new clean and disinfectant solution for 30 minutes, take the muscle cells of the hind legs aseptically, cut them into pieces, and contain 100U / ml card Wash twice with the MEM culture solution of Namycin, remove the washing solution; add 0.2% (w / v) collagenase II and 0.25% (w / v) trypsin digestion solution of 10 times the weight of muscle tissue, and place at 4°C Digestion: Discard the digestive fluid after 16-20 hours, add MEM solution 10 times the mass of muscle tissue to repeatedly blow and disperse the cells, pass through 100, 200 and 400 mesh stainless steel filters in turn, collect the filtrate, centrifuge, discard the supernatant, and precipitate the cells Resuspend with MEM, add newborn bovine serum to a final concentration of 10% (v / v), inoculate cell bottles, and ...

Embodiment 2

[0035] Embodiment 2: Preparation of CoxA16 vaccine

[0036] In Example 1, the inoculated virus was replaced with the CoxA16 virus strain (provided by Hangzhou Sixth People's Hospital), and the other preparation methods were the same, and CoxA16 vaccine semi-finished products and finished vaccines could be prepared.

Embodiment 3

[0037] Embodiment 3: Preparation of EV71 and CoxA16 bivalent vaccine

[0038] The EV71 vaccine semi-finished product prepared in Example 1 and the CoxA16 vaccine semi-finished product prepared in Experimental Example 2 were mixed at a volume ratio of 1:1 and subpackaged to obtain a bivalent enterovirus vaccine.

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PUM

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Abstract

The invention provides a method for preparing enterovirus vaccines, which uses gerbil muscle cells as stroma cells of virus culture. The method uses the gerbil muscle cells as the stroma cells for culturing EV71 or CoxA16 viruses, growth titer of the EV71 or CoxA16 viruses on the cells is high, and liquid collection can be conducted repeatedly. The cells can be performed with passage repeatedly, muscle cell strains can be built for production of vaccines, and consumption of gerbil is saved. The cells are primary cells so that the problems that passage cell deoxyribonucleic acids (DNA) are difficult to process and the like do not exist. The vaccine production process is relatively easy, purification is convenient, and the prepared vaccines are safe and reliable when used for vaccination of infants.

Description

(1) Technical field [0001] The invention relates to a preparation method of an enterovirus vaccine. (2) Background technology [0002] HFMD can be caused by a variety of enteroviruses, including CoxA5, A10, A16, A19, EV71, and some echoviruses and Coxsackie group B viruses, among which CoxA16 and EV71 are the most common. [0003] Enterovirus 71 (EV71) belongs to the genus Human Enterovirus, Picornaviridae, and is one of the main pathogens of hand, foot and mouth disease. Hand, foot, mouth and herpetic angina after infection in infants and young children, a small number of complications such as aseptic meningitis, brainstem encephalitis, neurogenic pulmonary edema and acute flaccid paralysis, severe nervous system, respiratory system and circulatory symptoms. Since the virus was first isolated in California, USA in 1969, the world has experienced many outbreaks of EV71. Since 1997, the number of people infected with EV71 has increased significantly, especially in the Asia...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/125A61K39/295A61P31/14C12N5/071
Inventor 朱函坪朱智勇夏时畅钱磊徐芳姚苹苹杨章女谢荣辉周小龙
Owner ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION
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