CHO (Chinese hamster ovary) cell serum-free medium supporting high expression of product

A serum-free medium and cell technology, which is applied in the direction of cell culture active agents, animal cells, vertebrate cells, etc., can solve the problems of difficult large-scale application of serum-free medium, high price and high cost, and achieve biosafety Sexual problems, prolonged plateau, and high expression

Active Publication Date: 2019-02-15
YOCON BIOLOGY TECH CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in the market, major media companies including imported and domestic brands have CHO serum-free media products, but th

Method used

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  • CHO (Chinese hamster ovary) cell serum-free medium supporting high expression of product
  • CHO (Chinese hamster ovary) cell serum-free medium supporting high expression of product
  • CHO (Chinese hamster ovary) cell serum-free medium supporting high expression of product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] Example 1 Preparation of serum-free medium for CHO cells

[0096] The concentration of each raw material is as follows:

[0097] Substance name Added concentration (mg / L)

[0098] amino acid

[0099] Glycine 650

[0100] L-alanine 46

[0101] L-Arginine Hydrochloride 324

[0102] L-Asparagine 850

[0103] L-Aspartic Acid 240

[0104] L-cysteine ​​hydrochloride 118

[0105] L-glutamic acid 187

[0106] L-Histidine Hydrochloride 235

[0107] L-Isoleucine 220

[0108] L-Leucine 430

[0109] L-Lysine hydrochloride 166

[0110] L-Methionine 207

[0111] L-phenylalanine 122

[0112] L-proline 74.5

[0113] L-serine 256

[0114] L-threonine 156

[0115] L-tryptophan 153.3

[0116] Disodium L-tyrosine 306.8

[0117] L-valine 323

[0118] L-cystine 185

[0119] Hydroxyproline 2.79

[0120] vitamins

[0121] Choline chloride 155.5

[0122] Calcium D-pantothenate 6.21

[0123] Folic acid 1.97

[0124] Niacinamide 4.57

[0125] Pyridoxine hydrochloride 9.1...

Embodiment 2

[0178] Example 2 Preparation of serum-free medium for CHO cells

[0179] The concentration of each raw material is as follows:

[0180] Substance name Added concentration (mg / L)

[0181] amino acid

[0182] Glycine 500

[0183] L-alanine 30

[0184] L-Arginine HCl 500

[0185] L-Asparagine 505

[0186] L-Aspartic Acid 240

[0187] L-cysteine ​​hydrochloride 130

[0188] L-glutamic acid 155

[0189] L-histidine hydrochloride 290

[0190] L-Isoleucine 450

[0191] L-leucine 125

[0192] L-Lysine Hydrochloride 250

[0193] L-Methionine 125

[0194] L-phenylalanine 60

[0195] L-Proline 60

[0196] L-serine 350

[0197] L-Threonine 300

[0198] L-Tryptophan 215

[0199] Disodium L-Tyrosine 400

[0200] L-Valine 400

[0201] L-cystine 252

[0202] Hydroxyproline 3.3

[0203] vitamins

[0204] Choline chloride 215

[0205] Calcium D-pantothenate 7.5

[0206] Folic acid 3.2

[0207] Niacinamide 5.8

[0208] Pyridoxine hydrochloride 7.5

[0209] Riboflavin 2.1...

Embodiment 3

[0261] Example 3 Preparation of CHO cell serum-free medium

[0262] The concentration of each raw material is as follows:

[0263] Substance name Added concentration (mg / L)

[0264] amino acid

[0265] Glycine 650

[0266] L-alanine 30

[0267] L-Arginine HCl 300

[0268] L-Asparagine 1000

[0269] L-Aspartic Acid 150

[0270] L-cysteine ​​hydrochloride 150

[0271] L-Glutamic Acid 200

[0272] L-Histidine Hydrochloride 200

[0273] L-Isoleucine 320

[0274] L-Leucine 430

[0275] L-Lysine Hydrochloride 185

[0276] L-Methionine 200

[0277] L-Phenylalanine 150

[0278] L-Proline 60

[0279] L-serine 285

[0280] L-threonine 250

[0281] L-Tryptophan 200

[0282] Disodium L-tyrosine 423

[0283] L-Valine 405

[0284] L-cystine 155

[0285] Hydroxyproline 3.0

[0286] vitamins

[0287] Choline Chloride 150

[0288] Calcium D-pantothenate 5.8

[0289] Folic acid 3.3

[0290] Niacinamide 4.2

[0291] Pyridoxine hydrochloride 8.5

[0292] Riboflavin 1.8 ...

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Abstract

The invention discloses a CHO (Chinese hamster ovary) cell serum-free medium. The culture medium comprises main components such as amino acids, inorganic salt components, vitamins, trace elements, yeast hydrolysates, albumin and the like. The CHO cell serum-free medium disclosed by the invention does not contain any serum components and animal-derived components, and is increased in affinity compared with that of other serum-free mediums. Cells can be directly inoculated into the serum-free medium from a serum culture medium or other serum-free mediums and grow normally, thus eliminating a cumbersome domestication process. By adopting the CHO cell serum-free medium, the growth density and the viability of CHO cells are improved, the tolerance of the cells therein is increased, the growth platform maintenance time is prolonged, most importantly, the ability of CHO engineering cells to express foreign proteins in the CHO cell serum-free medium is improved, and the product yield is greatly enhanced.

Description

technical field [0001] The invention relates to a serum-free medium for CHO cell culture in vitro, belonging to the technical field of cell culture. Background technique [0002] Cell culture technology is also called cell cloning technology, which means that under in vitro conditions, by adding specific nutrients and controlling the ambient temperature, pH, osmotic pressure, etc., the cells can maintain activity and grow normally in a suitable vessel. According to the difference in the way cells adhere to the wall in the vessel, the culture methods can be divided into adherent culture and suspension culture. Cell culture medium is the source for cells to absorb nutrients and is the key to the success of cell culture. According to different sources, cell culture media can be divided into natural media and synthetic media. Natural media consist only of naturally occurring biological fluids. Natural media are very useful and convenient for many different animal cell culture...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/02
CPCC12N5/0682C12N2500/12C12N2500/14C12N2500/16C12N2500/20C12N2500/22C12N2500/25C12N2500/32C12N2500/34C12N2500/36C12N2500/38C12N2500/40C12N2500/46C12N2500/90C12N2501/39C12N2501/998
Inventor 闫攀登路明华
Owner YOCON BIOLOGY TECH CO
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