Method for preparing rabies vaccine stock solution from serum-free Vero cells and serum-free rabies vaccine product

Abstract

The invention provides a method for preparing a rabies vaccine stock solution from serum-free Vero cells. The method comprises the following steps: culturing Vero cells with a serum-free culture medium to obtain a serum-free culture medium adaptive cell line; establishing a serum-free Vero cell seed bank by utilizing the serum-free culture medium adaptive cell strain; recovering, culturing, performing passage and amplifying cells of the Vero cell working seed bank by using the serum-free culture medium to serve as basal cells for bioreactor culture, and continuously perfusing to culture high-density Vero cells with the serum-free culture medium by applying a bioreactor and a microcarrier after cell expansion; inoculating virus seeds of a rabies vaccine virus strain working seed bank, performing bioreactor microcarrier serum-free culture, starting to continuously perfuse to obtain a virus fluid when the virus is amplified to peak to obtain titer of the virus fluid; and performing clarifying, ultrafiltration concentration, inactivation and purification treatment to obtain the serum-free human rabies vaccine stock solution.

Description

technical field [0001] The invention relates to a preparation method of a rabies vaccine stock solution, in particular to a method for preparing a rabies vaccine stock solution by culturing serum-free Vero cells with bioreactor microcarriers and serum-free rabies vaccine products. Background technique [0002] At present, the cell substrates used in the mass production of human rabies vaccines include primary cells, such as primary hamster kidney cells (PHKCV) and primary chicken embryo fibroblasts (PCECV); continuous passage cells, such as African green monkey kidney cells (Vero) and human diploid cells, etc. 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...

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Problems solved by technology

[0007] 1. The production of vaccines with serum is unstable and the quality is difficult to control;

[0008] 2. Potential viral microorganisms in serum are unsafe;

[0010] 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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