Non-serum non-animal-origin-additive insect cell culture medium

A technology of insect cells and animal sources, applied in the direction of invertebrate cells, animal cells, cell culture active agents, etc., can solve the problems of high requirements for transportation and bacteria release, high price, unsuitable for the development of medium, etc., to achieve Short doubling time, low cost, and easy preparation

Inactive Publication Date: 2017-02-22
严志海
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the Chinese patent application (publication number CN1498267A) applied by Bashofer Co., Ltd., the medium produced by it is a liquid medium, which has extremely high requirements for transportation and bacteria release, and is expensive, and is not suitable for the current medium of China process status

Method used

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  • Non-serum non-animal-origin-additive insect cell culture medium
  • Non-serum non-animal-origin-additive insect cell culture medium
  • Non-serum non-animal-origin-additive insect cell culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1 A kind of serum-free insect cell culture medium without animal source added

[0022] A serum-free insect culture medium without animal-derived additives, consisting of the following components and their concentrations: Grace medium 7.5g / L, glucose 7.2g / L, sodium chloride 21.5mM, sodium dihydrogen phosphate 11mM, citric acid Sodium 5.5mM, sodium selenite 13.5mM, potassium chloride 6mM, potassium sulfate 5.23mM, aluminum chloride 2.75mM, zinc sulfate 14.5mM, manganese chloride tetrahydrate 3.2mM, ammonium molybdate 0.67mM, biotin 1.75mg / L, vitamin B 12 0.38mg / L, riboflavin 1.82mg / L, p-aminobenzoic acid 0.35mg / L, pantothenic acid 7.6mg / L, vitamin B 1 0.71mg / L, folic acid 0.27mg / L, choline chloride 17mg / L, L-arginine 5.6mM, L-asparagine 4.75mM, L-glycine 8.2mM, L-histidine 2.45mM, L -Isoleucine 8.65mM, L-Leucine 7.68mM, L-Lysine 8.15mM, L-Methionine 0.49mM, L-Threonine 7.2mM, L-Tryptophan 1.14mM, L-Kytidine Acid 3.32mM, L-proline 8.12mM, L-glutamine 13.75g / ...

Embodiment 2

[0028] Embodiment 2 A kind of serum-free insect cell culture medium without animal source added

[0029] A serum-free insect culture medium without animal-derived additives, consisting of the following components and their concentrations: basal medium 6g / L, glucose 6.5g / L, sodium chloride 20mM, sodium dihydrogen phosphate 10mM, sodium citrate 5mM, sodium selenite 12mM, potassium chloride 5mM, potassium sulfate 4mM, aluminum chloride 1mM, zinc sulfate 10mM, manganese chloride tetrahydrate 1mM, ammonium molybdate 0.3mM, malic acid 0.4g / L, fumaric acid 0.1g / L, biotin 0.15mg / L, vitamin B 12 0.3mg / L, riboflavin 1.5mg / L, p-aminobenzoic acid 0.21mg / L, pantothenic acid 7mg / L, vitamin B 1 0.69mg / L, folic acid 0.2mg / L, choline chloride 16mg / L, L-arginine 4mM, L-asparagine 3mM, L-glycine 7mM, L-histidine 1.5mM, L-isoleucine Glycine 8.5mM, L-Leucine 7mM, L-Lysine 4mM, L-Methionine 0.35mM, L-Threonine 4.5mM, L-Tryptophan 0.69mM, L-Gyptophan 2.87mM, L - Proline 7.38mM, L-Glutamine 7g / L,...

Embodiment 3

[0031] Embodiment 3 A kind of serum-free insect cell culture medium without animal source added

[0032] An insect culture medium without serum and animal-derived additives, consisting of the following components and their concentrations: basal medium 8g / L, glucose 8g / L, sodium chloride 23mM, sodium dihydrogen phosphate 12mM, sodium citrate 7mM , sodium selenite 15mM, potassium chloride 8mM, potassium sulfate 6mM, aluminum chloride 3.5mM, zinc sulfate 20mM, manganese chloride tetrahydrate 5mM, ammonium molybdate 0.7mM, malic acid 1.8g / L, fumaric acid 0.8g / L, biotin 2.75mg / L, vitamin B 12 0.42mg / L, riboflavin 2.3mg / L, p-aminobenzoic acid 0.42mg / L, pantothenic acid 8mg / L, vitamin B10.74mg / L, folic acid 0.3mg / L, choline chloride 18mg / L, L- Arginine 6mM, L-Asparagine 5mM, L-Glycine 10mM, L-Histidine 3mM, L-Isoleucine 9mM, L-Leucine 9mM, L-Lysine 9mM, L-Methionine 0.54 mM, L-threonine 7.5mM, L-tryptophan 1.2mM, L-glutine 4.32mM, L-proline 8.42mM, L-glutamine 14.5g / L, yeast extra...

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Abstract

The invention relates to the field of cell culture medium, and in particular to a non-serum non-animal-origin-additive insect cell culture medium. The medium comprises mainly the following components: basic culture medium, glucose, inorganic salt, vitamins, L-arginine, L-agedoite, L-glycocoll, L-histidine, L-isoleucine, L-leucine, L-lysine, L-methionine, L-threonine, L-tryptophan, L-valine, L-proline, L-glutamine, yeast extracts, recombulin, malic acid, allomaleic acid, cholesterol, linoleic acid, granulesten, putrescine, glutathione, glycerol and fructosan. The culture medium can promote the growth of insect cell and is suitable for the large scale breeding of insect cell and the expression of recombination protein.

Description

technical field [0001] The invention relates to the field of cell culture medium, in particular to a serum-free and animal-source-free insect cell culture medium. Background technique [0002] Sf-9 cells and HighFive cells are the host cells of the baculovirus expression system. It has many advantages in the application of protein expression, preparation and purification, including: the expressed recombinant protein can be folded correctly and form disulfide bonds; post-translational modification processing such as glycosylation, phosphorylation, amidation and signaling Peptide cleavage, etc., make the recombinant protein closer to the natural protein in structure and function; Compared with other eukaryotic expression systems, the baculovirus expression system can express foreign genes efficiently, and the expression amount can reach the total protein amount of infected insect cells 50%; can accommodate large fragments of foreign genes. [0003] At present, domestic produ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/07
CPCC12N5/0601C12N2500/05C12N2500/12C12N2500/22C12N2500/30C12N2500/32C12N2500/34C12N2500/35C12N2500/38C12N2500/74C12N2501/33C12N2501/998
Inventor 严志海
Owner 严志海
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