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Serum-free medium used for suspension culture of insect cells, and application thereof

A serum-free medium and suspension culture technology, applied in the field of medium, can solve the problems of high production cost, complicated preparation, and complicated preparation of medium, and achieve the effect of low cost, simple preparation, and stable acquisition

Pending Publication Date: 2016-08-17
北京纳百生物科技有限公司
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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 provide a serum-free medium for insect cell suspension culture, to overcome the problems of complicated preparation, complex preparation and high production cost of the existing medium

Method used

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  • Serum-free medium used for suspension culture of insect cells, and application thereof
  • Serum-free medium used for suspension culture of insect cells, and application thereof
  • Serum-free medium used for suspension culture of insect cells, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: the preparation of serum-free medium of the present invention

[0033] (1) weighing

[0034] Accurately weigh raw materials according to Table 1, Table 2, and Table 3:

[0035] Table 1 Amino Acids

[0036]

[0037] Table 2 Vitamins

[0038]

[0039] Table 3 Inorganic salts

[0040]

[0041] Table 4 Lipid emulsifier

[0042]

[0043] After weighing the ingredients listed in Table 4, dissolve the above ingredients with 1mL of ethanol, then slowly add 9mL of water dropwise under heating (45-50°C) to make it slowly form an emulsion, and add the culture medium before use.

[0044] Table 5 Other ingredients

[0045]

[0046] (2) dissolve

[0047] Put the raw materials (Table 1, Table 2, Table 3, Table 5) into a large beaker, put 800mL ultrapure water into the beaker, stir until it is completely dissolved, add the solution prepared in Table 4 into the beaker, and use 10M sodium hydroxide was used to adjust the pH of the medium to 6.3, and ul...

Embodiment 2

[0048] Embodiment 2: culture medium of the present invention compares with imported similar product culture effect

[0049] According to conventional methods, SF9 cells were cultured in suspension with self-made NB-SFM medium and SF900 III medium, and the culture effects were compared (attached figure 1 ). from figure 1 It can be seen that during the same culture period, the density of viable cells in the two media SF9 is the same, and there is no significant difference.

[0050] Also according to the conventional method, use the self-made NB-SFM medium and the commercial medium to culture High Five cells in suspension, and compare the culture effect (attached figure 2 ). from figure 2 It can be seen that during the same culture period, the density of viable cells in the two media High Five is the same, and there is no significant difference.

Embodiment 3

[0051] Example 3: Self-made medium and commercial medium for cultivating insect cells after infection

[0052] According to the conventional method, the baculovirus-infected SF9 cells were cultured for 96 hours using self-made medium and commercial serum-free medium, and then the virus titer was measured, and the culture effects of the two media were compared (Table 6). It can be seen from Table 6 that the virus titer of self-made serum-free medium was significantly higher than that of commercial medium.

[0053] Table 6 Comparison of virus titers after baculovirus-infected cells were cultured for 96h

[0054]

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PUM

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Abstract

A serum-free medium used for suspension culture of insect cells includes 5-10g / L of carbohydrates, 1-5g / L of amino acids, 5-10g / L of an inorganic salt, 0.01-0.05g / L of vitamins, 5-10g / L of yeast extract and 10-20ml / L of a lipid emulsifier. Compared with serum component-containing media, the serum-free medium has the advantages of low cost, easy preparation, no animal pollution, and high batch stability; and compared with general commercial media, the serum-free medium has the advantages of realization of suspension culture of cells, improvement of the expression level, suitableness for large-scale production application, and facilitation of stable obtaining of recombinant proteins.

Description

technical field [0001] The invention relates to the field of medium, and relates to a serum-free medium for insect cell suspension culture and application thereof. Background technique [0002] Cell culture refers to the propagation and cultivation of cells in vitro. Cell culture technology, which started in the early 20th century, has become an important tool in the field of medical and biological research. More and more biological drugs such as monoclonal antibodies, vaccines and active proteins are successfully expressed and produced by cells. In the last 20 years, invertebrate cell culture, represented by insect cells, has played an increasingly important role in the field of applied biomedicine. Using baculovirus as a vector, the insect cell expression system has been widely used in the production of foreign proteins and is favored by people. In addition, the insect cell expression system is also considered to be the best system for expressing virus-like particle vacc...

Claims

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Application Information

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IPC IPC(8): C12N5/07C12N5/02
CPCC12N5/0601C12N2500/05C12N2500/30C12N2500/32C12N2500/38C12N2500/74C12N2500/90
Inventor 杨春江赵荣茂马孝斌秦堃莫勋吴迪
Owner 北京纳百生物科技有限公司
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