Method for preparing rotavirus vaccine stock solution by using serum-free Vero cells and serum-free rotavirus vaccine product

Abstract

The invention provides a method for preparing a rotavirus vaccine stock solution by using serum-free Vero cells. The method comprises the following steps: culturing Vero cells by using a serum-free culture medium, thus obtaining serum-free culture medium adapted cell strains; establishing a serum-free Vero cell seed bank by utilizing the obtained serum-free culture medium adapted cell strains; establishing a serum-free rotavirus strain working seed bank by utilizing the obtained serum-free culture medium adapted cell strains; carrying out reviving, culturing, passage and amplification on cells in a Vero cell working seed bank by utilizing the serum-free culture medium, using the cells in the Vero cell working seed bank as basic cells cultured in a bioreactor, and carrying out continuous perfusion culture on high-density Vero cells by applying the bioreactor and a microcarrier and using the serum-free culture medium after cell amplification; after inoculating virus seeds in the rotavirus strain working seed bank, carrying out bioreactor-microcarrier serum-free culture, obtaining a virus solution when virus is amplified to the summit, obtaining liquid virus titer, and carrying out clarification and ultra-filtration concentration, thus obtaining a serum-free rotavirus stock solution for human.

Description

technical field [0001] The invention relates to a preparation method of a rotavirus vaccine stock solution, in particular to a method for preparing a rotavirus vaccine stock solution by cultivating serum-free Vero cells with bioreactor microcarriers and serum-free rotavirus vaccine products. Background technique [0002] Currently, the cell substrates used in the mass production of human rotavirus vaccines include primary cells, such as calf kidney cells, and continuous passage cells, such as African green monkey kidney cells (Vero). Vero cells are a vaccine production cell line recognized by the World Health Organization. Compared with primary cells, diploid cells and some other passage cell matrices used for vaccine production, Vero cells have the characteristics of continuous passage, fast growth, stable genetic traits, low degree of malignant transformation, high biological safety, and culture The conditions are not harsh, and it is easy to implement large-scale cultiva...

AI Technical Summary

Problems solved by technology

[0009] 1. The production of serum vaccines is unstable and the quality is difficult to control;

[0010] 2. Potential viral microorganisms in serum will cause unsafe hidden dangers;

Especially for some viruses (such as rotavirus, etc.) Virus titer of harvested virus fluid

[0012] 4. Square bottles, spinner bottles, and cell factories are used as culture containers, which cannot control the process parameters well, which is not conducive to the gas exchange and sufficient nutrient supply during the cell growth process, and cannot control the process well, and cannot achieve high cell density. It is also difficult to achieve product uniformity when cultivating and obtaining higher titer virus fluid

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