Method for culturing baby hamster kidney (BHK) 21 cell in serum-free way, and vaccine preparation method

A serum-free culture, cell technology, applied in microorganism-based methods, animal cells, vertebrate cells, etc., can solve the problems of high cell culture density, poor safety, low cost, etc., to achieve higher cell culture density and batch differences. The effect of reducing performance and improving efficiency

Inactive Publication Date: 2011-07-06
BEIJING SKYWING TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to overcome the shortcomings of low cell culture density, high cost of downstream products of isolated cells, large batch differences of cultured cells and poor safety in the existing method for culturing BHK21 cells

Method used

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  • Method for culturing baby hamster kidney (BHK) 21 cell in serum-free way, and vaccine preparation method
  • Method for culturing baby hamster kidney (BHK) 21 cell in serum-free way, and vaccine preparation method
  • Method for culturing baby hamster kidney (BHK) 21 cell in serum-free way, and vaccine preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] This example is used to illustrate the animal cell culture method of the present invention.

[0063] table 3

[0064] components

g / 100L

components

g / 100L

Calcium Chloride Anhydrous

20

L-tryptophan

0.5

copper sulfate pentahydrate

0.0001

L-Tyrosine

7

Ferric nitrate nonahydrate

0

L-valine

3

Ferrous Sulfate Heptahydrate

0.2

D-glucose

550

potassium chloride

30

HEPES

400

Magnesium chloride hexahydrate

1

Linoleic acid

0.03

Anhydrous Magnesium Sulfate

6

lipoic acid

0.05

Sodium chloride

600

1,4-Butanediamine dihydrochloride

0.7

Anhydrous Sodium Dihydrogen Phosphate

5

sodium pyruvate

20

Disodium phosphate

6

Vitamin H

0.001

Zinc sulfate heptahydrate

0.04

Calcium D-pantothenate

0.4

Potassium dihydrogen phosphate anhydrous

8...

Embodiment 2

[0069] This example is used to illustrate the animal cell culture method of the present invention.

[0070] Table 4

[0071] components

g / 100L

components

g / 100L

Calcium Chloride Anhydrous

10

L-tryptophan

2

[0072] copper sulfate pentahydrate

0

L-Tyrosine

10

Ferric nitrate nonahydrate

0.01

L-valine

10

Ferrous Sulfate Heptahydrate

0.1

D-glucose

700

potassium chloride

20

HEPES

330

Magnesium chloride hexahydrate

10

Linoleic acid

0.05

Anhydrous Magnesium Sulfate

4

lipoic acid

0.08

Sodium chloride

620

1,4-Butanediamine dihydrochloride

0.02

Anhydrous Sodium Dihydrogen Phosphate

7

sodium pyruvate

10

Disodium phosphate

10

Vitamin H

0.005

Zinc sulfate heptahydrate

0.01

Calcium D-pantothenate

0.2

Potassium dihydrogen phospha...

Embodiment 3

[0077] This example is used to illustrate the animal cell culture method of the present invention.

[0078] table 5

[0079] components

g / 100L

components

g / 100L

Calcium Chloride Anhydrous

17

L-tryptophan

1

copper sulfate pentahydrate

0.001

L-Tyrosine

2

[0080] Ferric nitrate nonahydrate

0.008

L-valine

5

Ferrous Sulfate Heptahydrate

0.5

D-glucose

600

potassium chloride

50

HEPES

370

Magnesium chloride hexahydrate

6

Linoleic acid

0.02

Anhydrous Magnesium Sulfate

8

lipoic acid

0.1

Sodium chloride

670

1,4-Butanediamine dihydrochloride

0.9

Anhydrous Sodium Dihydrogen Phosphate

8

sodium pyruvate

11

Disodium phosphate

7

Vitamin H

0.002

Zinc sulfate heptahydrate

0.02

Calcium D-pantothenate

0.1

Potassium dihydrogen phosphat...

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Abstract

The invention provides a method for culturing baby hamster kidney (BHK) 21 cell in a serum-free way, which leads the BHK 21 cell to be inoculated into a cell culture medium for culturing, wherein the cell culture medium comprises a basic culture medium and further comprises 100-200g/100L of soy protein, 100-200g/100L of pea protein, 100-200g/100L of broad bean protein, 0-100g/100L of potato protein, 0-200g/100L of wheat gluten protein and 50-100g/100L of rice protein by taking the volume of solvent of the culture medium as reference. In addition, the invention also provides a vaccine (such asrabies vaccine and foot-and-mouth disease vaccine) preparation method comprising the method for culturing the BHK 21 cell. The BHK 21 cell cultured by the culture medium containing the vegetable protein is high in culture density, beneficial to separating down-stream products of the cells, low in cost, small in batch difference, good in safety and suitable for producing virus host, expression vector and the like of biological products such as vaccine and the like.

Description

technical field [0001] The invention relates to a method for cultivating BHK21 cells and a method for preparing a vaccine, in particular to a method for cultivating BHK21 cells without serum and a method for preparing a vaccine comprising the method for cultivating BHK21 cells. Background technique [0002] In the existing methods for culturing BHK21 cells without serum, soybean hydrolyzate is generally added as a component of serum-free animal cell culture medium to improve the stability and culture density of cultured BHK21 cells, increase the yield and utilization of cultured BHK21 cells Yield of recombinant target protein products produced by BHK21 cells as expression vectors. And it is believed that hydrolysates of other plants, such as wheat hydrolysates, do not obtain comparable beneficial effects as soybean hydrolysates. [0003] For example, CN 1318577C (Patent No. 00816020.1) discloses a method for cultivating mammalian cells, which uses a protein-free and serum-f...

Claims

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Application Information

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IPC IPC(8): C12N5/071A61K39/205A61K39/135A61P31/14C12N7/00C12R1/93
Inventor 张韧陈文庆王建超
Owner BEIJING SKYWING TECH CO LTD
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