Preparation method of novel dual-adjuvant coronavirus inactivated vaccine and application thereof

A coronavirus and inactivated vaccine technology, applied in biochemical equipment and methods, viruses, antiviral agents, etc., can solve the problems of insufficient immune effect and immune time, low neutralizing antibody value, etc., to improve the ability of autoimmune response , clear ingredients, and the effect of improving the stability of the vaccine

Pending Publication Date: 2021-08-27
ZHEJIANG TIANYUAN BIO PHARM CO LTD +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to the animal and clinical test data of several new coronavirus inactivated vaccines that are currently developing r

Method used

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  • Preparation method of novel dual-adjuvant coronavirus inactivated vaccine and application thereof
  • Preparation method of novel dual-adjuvant coronavirus inactivated vaccine and application thereof
  • Preparation method of novel dual-adjuvant coronavirus inactivated vaccine and application thereof

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preparation example Construction

[0016] The present invention relates to a kind of preparation method of novel adjuvant novel coronavirus inactivated vaccine, described preparation method comprises the following steps:

[0017] ① Preparation of cell matrix and virus fluid for production;

[0018] ② Virus inactivation and verification;

[0019] ③ Preparation of inactivated vaccines;

[0020] ④ Preparation of semi-finished products and finished products.

Embodiment 1

[0021] Embodiment 1: Preparation of cell matrix and virus liquid for production:

[0022] ① Cell resuscitation: Take out the corresponding number of Vero cells (purchased from ATCC, number CCL-81) from the liquid nitrogen tank, and after checking, resuscitate them in a T25 cell culture bottle (purchased from Corning), and the medium contains 20% neonatal The 199 medium of bovine serum (the 199 medium was purchased from Beijing Tianxinhe Biotechnology, and the newborn bovine serum was purchased from Hangzhou Tianhang Biology) was placed in a constant temperature incubator at 37°C for cultivation until the cells filled a dense monolayer.

[0023] ②Cell passage: Passage the Vero cells covered with a dense monolayer, wash the cells slowly with an appropriate amount of 0.01M PBS solution (homemade), then add an appropriate amount of 0.25% trypsin digestion solution (purchased from Gbico) for digestion, and mix well into a cell suspension. Divide the cell suspension into correspond...

Embodiment 2

[0026] Embodiment 2: Harvesting and inactivation of virus liquid

[0027] ① Observe the lesions of the cell factory. After the lesions of the cell factory reach 50%-80%, put the cell factory into the refrigerator at 2-8°C to pre-cool to 4-8°C, and then add the diluted β- Propanolactone (purchased from SERVA) was placed at 2-8°C for further inactivation for 20 hours.

[0028] ② After the inactivation is completed, according to the total amount of harvested liquid and the liquid volume in the collection bucket, the inactivation sampling is carried out at a liquid volume of not less than 0.1% in each batch. The sample volume is sampled according to different harvesting containers, inoculated into Vero cells, and blind Passage 3 generations, pass 1 generation every 4 to 5 days, and check the virus by cytopathic method for each generation, and the results should all be negative.

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Abstract

The invention discloses a preparation method of a novel double-adjuvant coronavirus inactivated vaccine and an application thereof, and the specific steps are as follows: after a novel coronavirus inactivated stock solution is prepared according to the same specified protein content or antigen content, a CpG adjuvant and an aluminum adjuvant are added, the total protein content of a novel coronavirus antigen is 2-4 [mu] g/ml, the CpG content is 5-40 [mu] g/ml, and the aluminum content is 450 [mu] g/ml. The method has the beneficial effects that the novel double-adjuvant coronavirus inactivated vaccine prepared by the method can effectively improve the serum titer and the neutralizing antibody level of the current novel aluminum adjuvant coronavirus inactivated vaccine; meanwhile, the antigen dosage is effectively reduced, the production cost is reduced, the vaccine yield and the use coverage rate of future global new crown vaccines are improved, and benefits are brought to human beings.

Description

technical field [0001] The invention relates to the field of inactivated vaccines, and mainly relates to a preparation method and application of a double-adjuvanted novel coronavirus inactivated vaccine. Background technique [0002] The outbreak of severe acute respiratory syndrome occurred at the end of December 2019, and the results of etiological testing confirmed that it was a new type of coronavirus, and then the cells were isolated to obtain a new type of coronavirus strain. The International Committee on Taxonomy of Viruses named the novel coronavirus as severe acute respiratory syndrome coronavirus 2 (severe acute respiratory syndrome coronavirus 2, SARS-CoV-2), and the WHO named the disease caused by the virus as novel coronavirus pneumonia (coronavirus disease 2019, COVID-19). -19), belonging to the betacoronavirus family. This is the seventh coronavirus known to infect humans; four coronaviruses (229E, NL63, OC43, and HKU1) cause only mild cold symptoms. In con...

Claims

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Application Information

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IPC IPC(8): A61K39/215A61K39/39A61P31/14A61P37/04
CPCA61K39/12A61K39/39A61P31/14A61P37/04A61K2039/5252A61K2039/55505A61K2039/55561C12N2770/20034
Inventor 沈吉友许定花陈燕王瑞博刘家胜张弓李方仲柯江伟龚昱彬邵彦邵聪文张翊
Owner ZHEJIANG TIANYUAN BIO PHARM CO LTD
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