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CHO (Chinese Hamster Ovary) cell serum-free medium

A serum-free culture medium and cell technology, applied in the field of cell culture medium, can solve the problems of increased difficulty, contamination of target protein separation and purification, etc.

Inactive Publication Date: 2018-04-06
成都源泉生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The technical problem to be solved by the present invention is that all the CHO cell culture medium in the prior art contains serum, but the addition of serum components brings exogenous pollution or increases the difficulty of separation and purification of the target protein. The purpose is to provide a CHO cell Serum-free medium does not contain serum components at all, which is beneficial to the protein purification process; in the CHO culture of this medium, CHO cells do not need to go through the adaptation process of reducing serum; it can maintain cell activity for a long time, prolong the production time of protein expression, and greatly Significantly reduces the cost of serum-free medium, reducing production costs for enterprises

Method used

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  • CHO (Chinese Hamster Ovary) cell serum-free medium

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Effect test

Embodiment 1

[0016] A serum-free medium for CHO cells, including raw materials at the following concentrations: alanine 50mg / L, arginine 40mg / L, aspartic acid 30mg / L, asparagine 50mg / L, cystine 40mg / L L, cysteine ​​70mg / L, glutamine 600mg / L, glutamic acid 20mg / L, glycine 30mg / L, histidine 50mg / L, hydroxyproline 70mg / L, isoleucine 80mg / L L. Leucine 30mg / L, Lysine 90mg / L, Methionine 60mg / L, Phenylalanine 50mg / L, Proline 40mg / L, Serine 50mg / L, Threonine 40mg / L, Tryptophan Acid 30mg / L, Tyrosine 50mg / L, Valine 30mg / L, Calcium Chloride 40mg / L, Magnesium Sulfate 35mg / L, Potassium Chloride 700mg / L, Potassium Dihydrogen Phosphate 60mg / L, Dihydrogen Phosphate Sodium 40mg / L, sodium bicarbonate 1g / L, sodium hydroxide 3g / L, ferrous sulfate heptahydrate 40mg / L, magnesium chloride hexahydrate 70mg / L, copper sulfate 30mg / L, sodium pyruvate 40mg / L, dichloro Manganese chloride 50mg / L, sodium butyrate 70mg / L, ferric citrate 20mg / L, citric acid 55mg / L, sodium chloride 3g / L, biotin 40mg / L, choline chloride 25...

Embodiment 2

[0018] A serum-free medium for CHO cells, including raw materials at the following concentrations: alanine 10mg / L, arginine 10mg / L, aspartic acid 10mg / L, asparagine 10mg / L, cystine 10mg / L L, cysteine ​​10mg / L, glutamine 100mg / L, glutamic acid 10mg / L, glycine 10mg / L, histidine 10mg / L, hydroxyproline 10mg / L, isoleucine 10mg / L L. Leucine 10mg / L, Lysine 10mg / L, Methionine 10mg / L, Phenylalanine 10mg / L, Proline 10mg / L, Serine 10mg / L, Threonine 10mg / L, Tryptophan Acid 10mg / L, Tyrosine 10mg / L, Valine 10mg / L, Calcium Chloride 10mg / L, Magnesium Sulfate 10mg / L, Potassium Chloride 10mg / L, Potassium Dihydrogen Phosphate 10mg / L, Dihydrogen Phosphate Sodium 10mg / L, sodium bicarbonate 0.1g / L, sodium hydroxide 0.1g / L, ferrous sulfate heptahydrate 10mg / L, magnesium chloride hexahydrate 10mg / L, copper sulfate 10mg / L, sodium pyruvate 10mg / L, Manganese dichloride 10mg / L, sodium butyrate 10mg / L, ferric citrate 10mg / L, citric acid 10mg / L, sodium chloride 1g / L, biotin 1mg / L, choline chloride 1mg / L, ...

Embodiment 3

[0020]A serum-free medium for CHO cells, including raw materials at the following concentrations: alanine 100mg / L, arginine 1100mg / L, aspartic acid 100mg / L, asparagine 100mg / L, cystine 100mg / L L, cysteine ​​100mg / L, glutamine 1000mg / L, glutamic acid 100mg / L, glycine 100mg / L, histidine 100mg / L, hydroxyproline 100mg / L, isoleucine 100mg / L L. Leucine 100mg / L, Lysine 100mg / L, Methionine 100mg / L, Phenylalanine 1100mg / L, Proline 100mg / L, Serine 100mg / L, Threonine 100mg / L, Tryptophan Acid 100mg / L, Tyrosine 100mg / L, Valine 100mg / L, Calcium Chloride 100mg / L, Magnesium Sulfate 100mg / L, Potassium Chloride 1000mg / L, Potassium Dihydrogen Phosphate 100mg / L, Dihydrogen Phosphate Sodium 100mg / L, sodium bicarbonate 5g / L, sodium hydroxide 5g / L, ferrous sulfate heptahydrate 100mg / L, magnesium chloride hexahydrate 100mg / L, copper sulfate 100mg / L, sodium pyruvate 100mg / L, dichloro Manganese chloride 100mg / L, sodium butyrate 100mg / L, ferric citrate 100mg / L, citric acid 100mg / L, sodium chloride 10g / ...

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Abstract

The invention discloses a serum-free medium for CHO cells, which comprises the following raw materials: alanine, arginine, aspartic acid, asparagine, cystine, cysteine, glutamine, glutamine Acid, glycine, histidine, etc., calcium chloride, magnesium sulfate, potassium chloride, potassium dihydrogen phosphate, disodium hydrogen phosphate, sodium bicarbonate, sodium hydroxide, ferrous sulfate heptahydrate, magnesium chloride hexahydrate, copper sulfate , sodium pyruvate, manganese dichloride, etc., nicotinamide, p-aminobenzoic acid, folic acid, ribose, uracil, lipoic acid, linoleic acid L, cholesterol, dexamethasone, yeast hydrolyzate, milk protein hydrolyzate, casein protein hydrolyzate. The medium of the present invention does not contain serum components at all, which is beneficial to the purification process of proteins; CHO cells do not need to go through the adaptation process of reducing serum; they can maintain cell activity for a long time, prolong the production time of protein expression, and greatly reduce the cost of serum-free medium. Cost, reduce production cost for enterprises.

Description

technical field [0001] The invention relates to a cell culture medium, in particular to a serum-free culture medium for CHO cells. Background technique [0002] After decades of development, cell culture is now widely used in biology, medical technology, pharmaceuticals and other fields, forming a huge industry. Products are becoming more and more subdivided to meet the needs of different purposes. Obtaining protein drugs and antibodies through animal engineering cell culture has become an important direction for the development of modern medicine. Half of the world's recombinant drugs are expressed in mammalian cells, and 70% of them are expressed in Chinese hamster ovary cells (CHO cells). It can be seen that CHO is the most important host cell for recombinant drug production. In the process of cultivating CHO cells to produce drugs, most of them use the addition of serum and protein components, and the addition of these components may often bring external pollution or ...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0682C12N2500/12C12N2500/14C12N2500/16C12N2500/20C12N2500/24C12N2500/32C12N2500/34C12N2500/36C12N2500/38C12N2500/40C12N2500/44C12N2500/46C12N2500/90C12N2501/33C12N2501/39C12N2501/392C12N2501/998C12N2501/999
Inventor 刘才明
Owner 成都源泉生物科技有限公司
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