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Serum-free and animal-source-free culture medium suitable for insect cell Sf-9

A technology of insect cells and medium, applied in the field of preparation of serum-free and animal-free medium, can solve the problems of high price, high cost, unsuitable medium development status, etc., and achieve the effect of promoting proliferation speed and high vigor

Inactive Publication Date: 2016-01-20
内蒙古金源康生物工程股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, the price of foreign serum-free medium products is high and the formula is confidential. At the same time, there is a lack of serum-free medium specifically for the cultivation of insect cell Sf-9 cells at home and abroad, and the performance of the medium is poor, and the vitality of the cells cannot be maintained continuously. or higher cost
The Chinese Patent Application Publication No. CN1498267A applied by Bassow AG, the culture medium produced by it is a liquid culture medium, which has extremely high requirements for transportation and bacteria release, and is expensive, which is not suitable for the current development of culture medium in my country

Method used

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  • Serum-free and animal-source-free culture medium suitable for insect cell Sf-9
  • Serum-free and animal-source-free culture medium suitable for insect cell Sf-9
  • Serum-free and animal-source-free culture medium suitable for insect cell Sf-9

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] Example 1. Preparation of serum-free, animal-free, low-protein medium

[0100] This example is a serum-free, animal-derived low-protein medium (JYK-Sf9) suitable for suspension culture of Sf-9 cells, which includes 22 amino acids, 12 inorganic salts, 12 vitamins, 6 lipids, and 3 Two kinds of protein hydrolysates, 12 kinds of additives; all materials used are purchased from sigma company unless otherwise specified. The specific ingredients are as follows:

[0101] (1) Amino acid part:

[0102] L-alanine 100-500mg / L,

[0103] L-glutamine 900-5500mg / L,

[0104] L-arginine hydrochloride 700-2500mg / L,

[0105] L-aspartic acid 900-1700mg / L,

[0106] L-Asparagine 800-1800mg / L,

[0107] L-cysteine ​​monohydrate 120-320mg / L,

[0108] L-glutamic acid 1300-2800mg / L,

[0109] L-Isoleucine 400-1300mg / L,

[0110] L-Leucine 300-750mg / L,

[0111] L-lysine hydrochloride 400-1600mg / L,

[0112] L-methionine 600-1650mg / L,

[0113] L-phenylalanine 700-1800mg / L,

[0114] L-proline 300-1000mg / L,

[0115] L-seri...

Embodiment 2

[0182] Example 2. Comparative experiment of using the medium of the present invention in cell culture

[0183] The suspension culture type Sf-9 cells are 100*10 4 The initial test density of Cells / ml was inoculated into a 1000ml cell triangle culture flask, and the culture conditions were: temperature 27 degrees Celsius, rotation speed 110 rpm / min, sampling and counting every 24 hours and checking cell viability. In the cell culture experiment, two groups were designed as control experiments. The experimental group was the self-made medium JYK-SF9, the control group 1 was the mature Gibico insect medium SF900II, and the control group 2 was the existing commercially available product. We chose Suzhou Womei Sf-9 cell culture medium serum-free medium SF-SFM, each group cultured 20 flasks of cells, repeated three times.

[0184] The results showed that the growth time of Sf-9 cells in the self-made medium JYK-SF9 was shorter, and the cell viability was significantly better than other c...

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Abstract

The invention relates to the technical field of biological pharmacy, in particular to preparation for a serum-free and animal-source-free culture medium suitable for an insect cell Sf-9. For the growth characteristic of the insect cell Sf-9, the low-cost, serum-free, animal-source-free and low-protein culture medium suitable for high-density suspension culture of the cell Sf-9 is developed. Components such as lipid, vitamins, microelement, recombinant protein, hydrolysate and the like are added to replace the biological function of serum, and some negative effects on later-stage purification and antigen separation from the serum are avoided. By optimizing the components of the culture medium, the proliferation speed of the cell Sf-9 in suspension culture is higher, cell density is higher, cell activity is higher, and the culture medium is more suitable for large-scale industrial production.

Description

Technical field [0001] The invention relates to the technical field of biopharmaceuticals, in particular to the preparation of a serum-free animal source-free culture medium suitable for insect cell Sf-9. Background technique [0002] The baculovirus expression system (baculovirus expression system) is a eukaryotic expression system that has been used more in recent years. It can express a variety of foreign genes in insect cells, including genes from fungi, plants, bacteria, and viruses. Baculovirus is a type of double-stranded DNA virus with insect cells as its natural host. It has high species specificity, does not infect vertebrates, and is harmless to humans and livestock. The main advantages of the baculovirus expression system are as follows: [0003] ① The expressed recombinant protein can fold correctly and form disulfide bonds; [0004] ②After translation, it can be modified and processed, such as glycosylation, phosphorylation, amidation and signal peptide cleavage, so t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/07
Inventor 赵峻岭苏广华黄新朋
Owner 内蒙古金源康生物工程股份有限公司
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