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Low-serum culture medium for Vero cell culture and corresponding virus production

A cell culture and low serum technology, which is applied in the direction of cell culture active agents, microorganism-based methods, viruses, etc., can solve the problems of cell culture medium pickiness, failure to meet requirements, and production requirements, etc., and achieve simple formula ingredients and improved The effect of cultivating the effect

Inactive Publication Date: 2019-12-27
山东巨山能源科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some commercially available low-serum medium cannot meet the production requirements, and cannot maintain the excellent growth state of cells under low-serum culture conditions, and at the same time maintain the stable toxicity of the semi-finished products of vaccine production
There are two production methods for Vero cell amplification of porcine epidemic diarrhea virus. The process of adding trypsin is more critical to the cell culture medium, and most commercially available low-serum medium cannot meet the requirements. Based on this, we propose a low-serum culture medium

Method used

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  • Low-serum culture medium for Vero cell culture and corresponding virus production
  • Low-serum culture medium for Vero cell culture and corresponding virus production
  • Low-serum culture medium for Vero cell culture and corresponding virus production

Examples

Experimental program
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Effect test

Embodiment 1

[0021] A low-serum medium for Vero cell culture and corresponding virus production, including the following components: glycine 22.38mg / L, alanine 6.48mg / L, arginine hydrochloride 225.25mg / L, asparagine monohydrate 59.00mg / L, aspartic acid 10.08mg / L, cysteine ​​hydrochloride monohydrate 8.78mg / L, cystine dihydrochloride 69.65mg / L, glutamic acid 10.43mg / L, glutamine 532.50mg / L L, histidine hydrochloride monohydrate 34.74mg / L, isoleucine 71.24mg / L, leucine 72.03mg / L, lysine hydrochloride 105.63mg / L, methionine 71.62mg / L, phenylpropanoid Amino acid 46.74mg / L, proline 13.88mg / L, serine 106.63mg / L, threonine 67.23mg / L, tryptophan 11.76mg / L, tyrosine disodium salt dihydrate 310.40mg / L , Valine 65.68mg / L, Hydroxyproline 9.95mg / L, Ornithine 25.00mg / L, Sodium Ascorbyl Phosphonate 2.50mg / L, Biotin 0.33mg / L, Choline Chloride 6.24mg / L L, D-calcium pantothenate 4.87mg / L, folic acid 3.08mg / L, nicotinamide 2.76mg / L, niacin 0.01mg / L, pyridoxine hydrochloride 1.11mg / L, pyridoxal hydrochloride...

Embodiment 2

[0024] A low serum medium for Vero cell culture and corresponding virus production, including the following components: glycine 15mg / L, alanine 5mg / L, arginine hydrochloride 200mg / L, asparagine monohydrate 50mg / L , Aspartic Acid 8mg / L, Cysteine ​​Hydrochloride Monohydrate 5mg / L, Cystine Dihydrochloride 50mg / L, Glutamic Acid 8mg / L, Glutamine 400mg / L, Histidine Hydrochloride Monohydrate 25mg / L, Isoleucine 60mg / L, Leucine 60mg / L, Lysine Hydrochloride 80mg / L, Methionine 50mg / L, Phenylalanine 40mg / L, Proline 10mg / L, Serine 80mg / L, threonine 50mg / L, tryptophan 10mg / L, tyrosine disodium salt dihydrate 250mg / L, valine 50mg / L, hydroxyproline 8mg / L, ornithine 20mg / L, sodium ascorbyl phosphonate 1mg / L, biotin 0.1mg / L, choline chloride 5mg / L, D-calcium pantothenate 1mg / L, folic acid 1mg / L, niacinamide 1mg / L, niacin 0.005mg / L, pyridoxine hydrochloride 1mg / L, pyridoxal hydrochloride 0.02mg / L, riboflavin 0.1mg / L, thiamine hydrochloride 10mg / L, vitamin B12 1mg / L, i-inositol 8mg / L L. Vita...

Embodiment 3

[0027] A low serum medium for Vero cell culture and corresponding virus production, including the following components: glycine 25mg / L, alanine 10mg / L, arginine hydrochloride 270mg / L, asparagine monohydrate 60mg / L , Aspartic Acid 11mg / L, Cysteine ​​Hydrochloride Monohydrate 10mg / L, Cystine Dihydrochloride 70mg / L, Glutamic Acid 15mg / L, Glutamine 600mg / L, Histidine Hydrochloride Monohydrate 50mg / L, Isoleucine 75mg / L, Leucine 75mg / L, Lysine Hydrochloride 110mg / L, Methionine 75mg / L, Phenylalanine 50mg / L, Proline 15mg / L, Serine 110mg / L, threonine 75mg / L, tryptophan 15mg / L, tyrosine disodium salt dihydrate 320mg / L, valine 75mg / L, hydroxyproline 12mg / L, ornithine 30mg / L, sodium ascorbyl phosphonate 5mg / L, biotin 0.5mg / L, choline chloride 7mg / L, D-calcium pantothenate 5mg / L, folic acid 5mg / L, niacinamide 5mg / L, niacin 0.01mg / L, pyridoxine hydrochloride 5mg / L, pyridoxal hydrochloride 0.08mg / L, riboflavin 0.5mg / L, thiamine hydrochloride 15mg / L, vitamin B12 5mg / L, i-inositol 10mg / L ...

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Abstract

The invention discloses a low-serum culture medium for Vero cell culture and corresponding virus production. The low-serum culture medium comprises the following constitution components: amino acids,vitamins, inorganic salts, proteins and auxiliary components, wherein the amino acids comprise glycine, arginine hydrochloride, glutamine, lysine hydrochloride, serine, disodium tyrosine dehydrate andvaline; and the proteins comprise whole albumin, lactoalbumin hydrolysate (LAH), saturated-iron human transferrin, whole-chain recombulin, 4-ethoxy piperazine ethanesulfonic acid (HEPES). In production of vaccines for porcine epidemic diarrhea viruses through Vero cell amplification, the culture medium disclosed by the invention is simple in formula component, low in cost and easy to prepare, anda good growth state of cells can be kept while cell culture effects are effectively improved.

Description

technical field [0001] The invention relates to the field of biopharmaceutical vaccine production, in particular to a low-serum culture medium for Vero cell culture and corresponding virus production. Background technique [0002] Vero cells are virus vaccine production cell lines approved by the World Health Organization and China Biological Products Regulations. The virus vaccines produced by vero cell culture technology mainly include rabies vaccine, polio vaccine, influenza vaccine, Japanese encephalitis vaccine, hemorrhagic fever vaccine and hand, foot and mouth disease vaccine, etc. Vero cells have wall-adhesive properties, which determines to a large extent that the production of virus vaccines is achieved through the immobilized culture of Vero cells on microcarriers. [0003] However, the traditional Vero cell culture method is to add 10% calf serum to the MEM medium to cultivate cells, which requires a large amount of serum and high cost. Reduced serum culture wi...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N7/00C12R1/93
CPCC12N5/0686C12N7/00C12N2500/32C12N2500/38C12N2500/12C12N2500/34C12N2500/40C12N2500/46C12N2501/33C12N2501/998C12N2770/24311
Inventor 齐智孟丹丹刘海英房圆瑗
Owner 山东巨山能源科技有限公司
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