HEK (human embryonic kidney) 293 cell line applicable to serum-free culture and application thereof

A serum-free culture and cell line technology, applied in the biological field, can solve the problems of relying on serum culture, low yield, and difficult purification.

Inactive Publication Date: 2012-07-25
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current adenovirus production process has disadvantages such as low yield, difficult purification, and dependence on serum culture.

Method used

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  • HEK (human embryonic kidney) 293 cell line applicable to serum-free culture and application thereof
  • HEK (human embryonic kidney) 293 cell line applicable to serum-free culture and application thereof
  • HEK (human embryonic kidney) 293 cell line applicable to serum-free culture and application thereof

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Experimental program
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Embodiment 1

[0036] 1 Materials and methods

[0037] 1.1 Cells, antibodies, media, viruses and other materials

[0038] The HEK293 cell line is preserved and provided by our laboratory; the monoclonal antibody HQ06 against classical swine fever virus E2 protein is prepared by our laboratory; serum-free culture medium 293 SFM II, DMEM, calf serum (FBS), GlutaMAX TM-1 were purchased from Life Technology Company; cell culture consumables were purchased from Corning Company; poly-lysine (Poly-L-Lysine, PDL) was purchased from Sigma Company; Recombinant adenovirus expressing the E2 gene of classical swine fever virus, the specific construction process is recorded in the application date of September 30, 2010, the application number is 201010297576.0, and the invention name is "adenovirus / alphavirus replicon chimeric vector swine fever vaccine and its application" in the patent literature.

[0039] 1.2 Culture method

[0040] Before the cells are adapted to culture, they are cultured with 10%...

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Abstract

The invention discloses an HEK (human embryonic kidney) 293 cell line applicable to serum-free culture and an application thereof. According to the invention, the HEK 293 cell (293SF) applicable to serum-free culture is obtained by adopting a culture solution progressive substitution method, wherein the preservation number of the HEK 293 cell is CGMCC (China General Microbiological Culture Collection Center) No.5824. Detection shows that the 293SF cell has stable adenovirus proliferation capacity and foreign protein expressing ability; average lesion time is 97 hours and is reduced by 12.9% compared with the HEK 293 cell; virus virulence reaches up to 107.48TCID50/mL and is improved by 43.3% compared with the HEK293 cell; and stability is good while generation number is improved, and the state and susceptibility of the 293SF cell are not changed after the 293SF cell is continuous passage is carried out for sixteen times. The cell line disclosed by the invention can be used for serum-free production of an adenovirus vector vaccine.

Description

technical field [0001] The invention relates to a cell line and application thereof, in particular to a HEK293 cell line adapted to serum-free culture and application thereof. Belongs to the field of biotechnology. Background of the invention [0002] Serum is a complex mixture formed by removing fibrin from plasma, and its components include carbohydrates, plasma proteins, polypeptides, fats, growth factors, hormones, inorganic substances, etc. These substances promote cell growth or inhibit cell growth activity so as to achieve physiological balance. In addition, serum provides hormones other than nutrients, growth factors, binding proteins that transport macromolecules, and contact-promoting and stretching factors that can promote cell attachment and protect them from mechanical damage. [0003] In routine cell culture, serum is an indispensable component, but there are still some problems. For example: the composition is complex, and there are obvious differences betw...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073C12N7/00C12P21/00C12R1/91
Inventor 仇华吉孙元
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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