Serum-free medium suitable for large-scale production of influenza vaccines

Abstract

The invention relates to the technical field of culture medium development and research of biotechnology and discloses a serum-free medium suitable for large-scale production of influenza vaccines. The serum-free medium comprises 23 basic metabolism nutrients, two nucleic acid compounds, 6 vitamins, 9 inorganic salt compounds, a shear force protective agent, two acidity and alkalinity buffer agents, an acidity and alkalinity indicator, 10 virus reproduction promoters and three additives. The conventional preparation method of the serum-free medium comprises the following steps of: dissolving the components in ultrapure water without a heat source so as to prepare the serum-free medium. The using method is a conventional method. The serum-free medium has the beneficial effects that the serum is not contained, the total protein content is lower than 10mg per liter, separation and purification of the product are promoted, and the serum-free medium is suitable for production of influenza vaccines, supports the normal growth and long-term continuous cell culture of animal cells and can be used without adaption; and moreover, because the components are clear, the serum-free medium is convenient to prepare, controllable in cost and suitable for large-scale production of the influenza vaccines.

Description

technical field [0001] The invention relates to the technical field of medium research and development of modern biotechnology, specifically, a serum-free medium suitable for large-scale microcarrier suspension culture of animal cells and virus amplification for vaccine production (large-scale production of influenza vaccine). technical background [0002] According to the data of the World Health Organization (WHO), approximately 5-15% of the world's population is infected with seasonal influenza each year, of which there will be 3-5 million severe cases and as many as 250-500,000 deaths. The losses caused by influenza to human beings are not only excess mortality, but also various forms of direct or indirect economic losses; among them, direct economic losses include hospital costs, increased medical consumption, and the efficiency of enterprises due to work absenteeism. Low and so on; someone calculated in 1999 that the annual direct economic loss caused by influenza to t...

AI Technical Summary

Problems solved by technology

However, this process is a labor-intensive process with many defects, for example: first, the requirements for chicken embryos are strict, and they must be SPF (Specific Pathogen Free) clean chicken embryos

Therefore, the period required to prepare qualified chicken embryos is long, and the ability to respond to influenza pandemics is poor

Second, individual differences in breeder chickens may lead to differences in chicken embryos, resulting in poor quality uniformity among batches of vaccines

Third, the production process of vaccines takes a long time and is prone to contamination

However, there may be many problems in the application of serum: for example: first, the difference in composition and quality between batches is likely to cause uneven quality between batches of biological products; second, it is easy to contaminate mycoplasma, virus, BSE-factor and other infectious agents. Third, the existence of a large amount of serum protein increases the difficulty of separation and purification in the later stage, and some substances cannot be completely removed, which will affect the quality of the final product; in addition, the source of serum is limited and expensive, which greatly increases the production cost

[0018] Although the above-mentioned commercial medium can support the stable growth of cells and be used for the production of biological products, there are also some obvious deficiencies. There will be many difficulties in large-scale cell culture and production of biological products

The main problems are: ①It is expensive and only suitable for some small-scale experimental research, not suitable for large-scale cell culture and production of biological products

② Most of the components are not disclosed, and the chemical composition is not clear, which brings certain difficulties to scientific research and production

③ Some media cannot support high-density production of cells, and some media have a long lag period when used for MDCK cell culture, and the growth of cells is slow, which increases production costs to a certain extent

④ Most of the medium can only support the adherent growth of MDCK cells, but not the proliferation of influenza virus. Even if it can support the proliferation of influenza virus, the obtained virus titer is not high

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