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

A serum-free medium and influenza vaccine technology, applied in the field of serum-free medium, can solve the problems of long cycle, inability to completely remove substances, unsuitable cell culture and production of biological products, etc.

Active Publication Date: 2013-04-17
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] A serum-free medium suitable for large-scale production of influenza vaccines, containing 23 basic metabolic nutrients, 2 nucleic acid compounds, 6 vitamins, 9 inorganic salt compounds, 1 shear protectant, 2 pH buffer, 1 pH indicator, 10 virus proliferation promoters and 3 additives, dissolve the following components in pyrogen-free ultrapure water, and the various components are as follows, the unit is mg / L:

[0050] Glucose (D-Glucose) 5400 Sodium Pyruvate 110 Alanine (L-Alanine) 4.45 Arginine (L-Arginine Hydrochloride) 147.5 Asparagine (L-Asparagine Monohydrate) 7.5 Aspartic acid (L-Aspartic cid) 6.65 Cystine (L-Cystine Dihydrochloride) 17.56 Cysteine ​​(L-Cysteine ​​Hydrochloride Monohydrate) 31.29 Glutamic Acid (L-Glutamic Acid) 7.35 Glutamine (L-Glutamine) 876 Glycine 18.75 Histidine (L-Histidine Hydrochloride Monohydrate) 31.48 Isoleucine (L-Isoleucine) 54.47 Leucine (L-Leucine) ...

Embodiment 2

[0054] A serum-free medium suitable for large-scale production of influenza vaccines, containing 23 basic metabolic nutrients, 2 nucleic acid compounds, 6 vitamins, 9 inorganic salt compounds, 1 shear protectant, 2 pH buffer, 1 pH indicator, 10 virus proliferation promoters and 3 additives, dissolve the following components in pyrogen-free ultrapure water, and the various components are as follows, the unit is mg / L:

[0055] Glucose (D-Glucose) 1000 Sodium Pyruvate 50 Alanine (L-Alanine) 4 Arginine (L-Arginine Hydrochloride) 40 Asparagine (L-Asparagine Monohydrate) 5 Aspartic acid (L-Aspartic cid) 2 Cystine (L-Cystine Dihydrochloride) 5 Cysteine ​​(L-Cysteine ​​Hydrochloride Monohydrate) 10 Glutamic Acid (L-Glutamic Acid) 3 Glutamine (L-Glutamine) 100 Glycine 5 Histidine (L-Histidine Hydrochloride Monohydrate) 10 Isoleucine (L-Isoleucine) 20 Leucine (L-Leucine) 30 Lysine (L-Lydine hydroc...

Embodiment 3

[0059] A serum-free medium suitable for large-scale production of influenza vaccines, containing 23 basic metabolic nutrients, 2 nucleic acid compounds, 6 vitamins, 9 inorganic salt compounds, 1 shear protectant, 2 pH buffer, 1 pH indicator, 10 virus proliferation promoters and 3 additives, dissolve the following components in pyrogen-free ultrapure water, and the various components are as follows, the unit is mg / L:

[0060] Glucose (D-Glucose) 6000 Sodium Pyruvate 200 Alanine (L-Alanine) 9 Arginine (L-Arginine Hydrochloride) 200 Asparagine (L-Asparagine Monohydrate) 20 Aspartic acid (L-Aspartic cid) 20 Cystine (L-Cystine Dihydrochloride) 50 Cysteine ​​(L-Cysteine ​​Hydrochloride Monohydrate) 50 Glutamic Acid (L-Glutamic Acid) 30 Glutamine (L-Glutamine) 2000 Glycine 50 Histidine (L-Histidine Hydrochloride Monohydrate) 50 Isoleucine (L-Isoleucine) 70 Leucine (L-Leucine) 80 Lysine (L-Lydin...

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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...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/02
Inventor 谭文松刘旭平赵亮范里黄锭
Owner EAST CHINA UNIV OF SCI & TECH
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