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Serum-free culture medium for hybridoma cell

A technology of serum-free medium and hybridoma cells, which is applied in the direction of cell culture medium, cell culture active agent, culture process, etc. It can solve the impact of product quality, the difficulty of separation and purification of target antibodies, and the complicated production process of hybridoma culture, etc. question

Inactive Publication Date: 2018-04-06
成都源泉生物科技有限公司
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  • 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 the cultivation and production process of hybridoma is relatively complicated, and the development of the most important part of the production process, the hybridoma culture medium, is very difficult. At present, the techniques adopted by most domestic manufacturers have animal serum added, and these The addition of components may often bring exogenous IgG, which will increase the difficulty of separating and purifying the target antibody and affect the quality of the product. The purpose is to provide a serum-free medium for hybridoma cells that does not contain serum components at all; the produced antibodies The purity is greatly improved; the medium has high amplification speed and high antibody expression ability at the same time; hybridoma cells do not need to be domesticated, and can directly transition from serum-containing culture to serum-free medium; this medium can maintain cells for a long time Activity, the culture time of hybridoma cells is greatly extended, which can greatly extend the production time and increase the antibody production

Method used

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  • Serum-free culture medium for hybridoma cell
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Examples

Experimental program
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Effect test

Embodiment 1

[0017] A serum-free medium for hybridoma cells, including raw materials at the following concentrations: alanine 50mg / L, arginine 40mg / L, aspartic acid 30mg / L, asparagine 50mg / L, cystine 40mg / L, cysteine ​​70mg / L, glutamine 600mg / L, glutamic acid 20mg / L, glycine 30mg / L, histidine 50mg / L, hydroxyproline 70mg / L, isoleucine 80mg / L, leucine 30mg / L, lysine 90mg / L, methionine 60mg / L, phenylalanine 50mg / L, proline 40mg / L, serine 50mg / L, threonine 40mg / L, color Amino 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, hydrogen phosphate Disodium 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, di Manganese chloride 50mg / L, sodium selenite 40mg / L, ammonium molybdate 5mg / L, sodium butyrate 70mg / L, sodium succinate 30mg / L, iron citrate 20mg / L, citr...

Embodiment 2

[0019]A serum-free medium for hybridoma cells, comprising raw materials at the following concentrations: alanine 10mg / L, arginine 10mg / L, aspartic acid 10mg / L, asparagine 10mg / L, cystine 10mg / L, cysteine ​​10mg / L, glutamine 100mg / L, glutamic acid 10mg / L, glycine 10mg / L, histidine 10mg / L, hydroxyproline 10mg / L, isoleucine 10mg / L, leucine 10mg / L, lysine 10mg / L, methionine 10mg / L, phenylalanine 10mg / L, proline 10mg / L, serine 10mg / L, threonine 10mg / L, color Amino 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, hydrogen phosphate Disodium 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 selenite 10mg / L, ammonium molybdate 0.001mg / L, sodium butyrate 10mg / L, sodium succinate 10mg / L, iron citrate 10mg / ...

Embodiment 3

[0021] A serum-free medium for hybridoma cells, including raw materials at the following concentrations: alanine 100mg / L, arginine 1100mg / L, aspartic acid 100mg / L, asparagine 100mg / L, cystine 100mg / L, cysteine ​​100mg / L, glutamine 1000mg / L, glutamic acid 100mg / L, glycine 100mg / L, histidine 100mg / L, hydroxyproline 100mg / L, isoleucine 100mg / L, leucine 100mg / L, lysine 100mg / L, methionine 100mg / L, phenylalanine 1100mg / L, proline 100mg / L, serine 100mg / L, threonine 100mg / L, color Amino 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, hydrogen phosphate Disodium 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, di Manganese chloride 100mg / L, sodium selenite 100mg / L, ammonium molybdate 100mg / L, sodium butyrate 100mg / L, sodium succi...

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Abstract

The invention discloses a hybridoma cell serum-free medium, which comprises the following raw materials: alanine, arginine, aspartic acid, asparagine, cystine, cysteine, glutamine, glutamic acid Amino 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, sulfuric acid Copper, sodium pyruvate, manganese dichloride, sodium selenite, ammonium molybdate, etc., nicotinamide, p-aminobenzoic acid, folic acid, ribose, uracil, lipoic acid, linoleic acid L, cholesterol, dexamethasone, Estradiol progesterone, testosterone, etc. The medium of the present invention does not contain serum components at all; the purity of the produced antibody is greatly improved; the medium has high amplification speed and high antibody expression ability; the medium can maintain cell activity for a long time, and the hybridoma cell culture time is greatly improved Extending can greatly extend the production time and increase the antibody production.

Description

technical field [0001] The invention relates to a cell culture medium, in particular to a serum-free culture medium for hybridoma 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. As a very important means of drug production, hybridoma cell culture to obtain antibodies has been widely used in the production of monoclonal antibodies and polyclonal antibodies. However, the cultivation and production process of hybridoma is relatively complicated. The most important part of the production process is the development of hybridoma medium, which is extremely difficult. At present, the process adopted by m...

Claims

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Application Information

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IPC IPC(8): C12N5/00
CPCC12N5/0037C12N2500/12C12N2500/14C12N2500/16C12N2500/20C12N2500/24C12N2500/32C12N2500/34C12N2500/36C12N2500/38C12N2500/40C12N2500/44C12N2500/46C12N2500/90C12N2501/33C12N2501/39C12N2501/392C12N2501/998C12N2501/999
Inventor 刘才明
Owner 成都源泉生物科技有限公司
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