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A kind of culture medium supplement, cell culture medium containing the supplement and application thereof

A cell culture and culture medium technology, applied in animal cells, cells modified by introducing foreign genetic material, vertebrate cells, etc., can solve the problems of decreased protein production, low yield, unstable genetic engineering cell lines, etc. The effect of expression level and high application value

Active Publication Date: 2022-03-18
BEIJING ASIA EAST BIO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current serum-free optimization for recombinant CHO cells expressing hepatitis B surface antigen has no obvious effect, and the output is still lower than the traditional production process
In addition, the process of serum-free acclimatization of cells may cause instability of genetically engineered cell lines, resulting in a decrease in protein production

Method used

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  • A kind of culture medium supplement, cell culture medium containing the supplement and application thereof
  • A kind of culture medium supplement, cell culture medium containing the supplement and application thereof
  • A kind of culture medium supplement, cell culture medium containing the supplement and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0034] This embodiment provides a kind of cell culture medium, in the DMEM medium that does not contain glutamine, add the component of following concentration:

[0035] 20mL / L newborn bovine serum, 0.1mg / mL proline, 0.1mg / mL asparagine and 0.1mg / mL histidine.

Embodiment 2

[0037] This embodiment provides a kind of cell culture medium, in the DMEM medium that does not contain glutamine, add the component of following concentration:

[0038] 20mL / L newborn bovine serum, 6μg / mL transferrin and 3μg / mL insulin.

Embodiment 3

[0040] This embodiment provides a kind of cell culture medium, in the DMEM medium that does not contain glutamine, add the component of following concentration:

[0041] 20 mL / L newborn bovine serum, 0.1 mg / mL proline, 0.1 mg / mL asparagine, 0.1 mg / mL histidine, 6 μg / mL transferrin and 3 μg / mL insulin.

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Abstract

The invention relates to a culture medium supplement, a cell culture medium containing the supplement and applications thereof, the culture medium supplement comprising proline, asparagine and histidine, and / or, transferrin and insulin . The above-mentioned medium supplements can be used for culturing CHO cells specifically expressing hepatitis B virus surface antigen by adding 20-30 mL / L newborn bovine serum to glutamine-free DMEM. The concentration of newborn calf serum used in the cell culture medium provided by the invention is as low as 2%, and it can be directly used for culturing CHO cells by adding specific components, without domesticating the CHO cells, and at the same time, the HBV surface antigen can be expressed in the CHO cells Levels rose, rising 16% to 40% in 13 days.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a medium supplement, a cell culture medium containing the supplement and applications thereof. Background technique [0002] Hepatitis B is a disease caused by hepatitis B virus (HBV). It is an infectious virus. Hepatitis B Surface Antigen (HBsAg) is used as a vaccine against hepatitis B virus. large recombinant proteins. At present, the expression systems of recombinant HBsAg mainly include yeast cells and Chinese hamster ovary cells (CHO). [0003] Chinese hamster ovary cells (CHO) transformed with HBV S protein gene can secrete and express recombinant HBsAg after cultured. Compared with the recombinant vaccine produced by yeast cells, it has better immunogenicity and positive conversion rate. The biological products expressed by animal cells, especially those expressed by CHO, are very safe and can be used in the production of hepatitis B vaccine without any serious side effects...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12P21/02
CPCC12N5/0682C07K14/005C12N2500/32C12N2500/25C12N2510/02C12N2730/10122C12N2730/10151
Inventor 付建家付立家张兰
Owner BEIJING ASIA EAST BIO PHARMA CO LTD
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