A kind of high-density fermentation method of recombinant human interferon alpha 2b

A recombinant human interferon and high-density fermentation technology, which is applied in the field of biomedicine, can solve the problems of low specific activity, bacterial aging, autolysis, and insufficient expression of foreign genes, so as to reduce the inhibitory effect of acetic acid and reduce acetic acid The generation and the effect of achieving high-density growth

Active Publication Date: 2012-02-01
BEIJING KAWIN TECH SHARE HLDG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The most serious ones are the premature aging of the bacteria during the culture process, the inability to fully express the autolysis and foreign genes, and the low specific activity. This is mainly due to the inhibition of the harmful metabolic waste produced by the bacteria on the recombinant bacteria.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 High-density fermentation of recombinant human interferon α2b

[0023] Fermentation medium 1: peptone 6g / L, yeast extract 2g / L, glucose 0.02%, glycerin 0.6%, Na 2 HPO 4 11g / L, NaH 2 PO 4 1g / L, KH 2 PO 4 4g / L, NaCl 2g / L, FeSO 4 .7H 2 O0.01g / L, MgSO 4 .7H 2 O 0.02g / L, CaCl 2 0.001g / L, VB 1 0.01g / L, MnSO 4 0.01g / L, glycine 0.2g / L, methionine 0.01g / L.

[0024] Feed medium includes: glucose concentration 15%, dosage 1%; glycerol concentration 45%, dosage 7%

[0025] (1) Take by weighing 100L of components according to the above formula;

[0026] (2) Dissolve in a specified amount of purified aqueous solution;

[0027] (3) adjust the pH value to 5.0-8.0 with sodium hydroxide solution;

[0028] (4) The fermentation medium is sterilized in a fermenter of corresponding volume (121°C, 30 minutes), and the feed medium is sterilized in a corresponding special glass feeding bottle (115°C, 30 minutes);

[0029] (5) Cultivate after inserting recombinan...

Embodiment 2

[0033] Embodiment 2: the high-density fermentation of recombinant human interferon alpha 2b

[0034] Fermentation medium 2: peptone 10g / L, yeast extract 5g / L, glucose 0.5%, glycerol 1%, Na 2 HPO 4 15g / L, NaH 2 PO 4 4g / L, KH 2 PO 4 6g / L, NaCl 3g / L, FeSO 4 .7H 2 O0.5g / L, MgSO 4 .7H 2 O 0.5g / L, CaCl 2 0.02g / L, VB 1 0.03g / L, MnSO 4 0.05g / L, glycine 0.5g / L, methionine 0.06g / L.

[0035] The feeding medium used in the present invention comprises: glucose concentration 35%, consumption 5%; glycerin concentration 25%, consumption 2%

[0036] (1) Take by weighing 100L of components according to the above formula;

[0037] (2) Dissolve in a specified amount of purified aqueous solution;

[0038] (3) adjust the pH value to 5.0-8.0 with sodium hydroxide solution;

[0039] (4) The fermentation medium was sterilized in a fermenter of corresponding volume (121° C., 30 minutes), and the feed medium was sterilized in a corresponding special glass feeding bottle (115° C., 3...

Embodiment 3

[0044] Embodiment 3: the high-density fermentation of recombinant human interferon alpha 2b

[0045] Fermentation medium 3: peptone 18g / L, yeast extract 8g / L, glucose 2%, glycerol 4%, Na 2 HPO 4 27g / L, NaH 2 PO 4 7.5g / L, KH 2 PO 4 6.5g / L, NaC16.5g / L, FeSO 4 .7H 2 O0.08g / L, MgSO 4 .7H 2 O 1.2g / L, CaCl 2 0.01g / L, VB 1 0.1g / L, MnSO 4 0.1gg / L, glycine 1.2g / L, methionine 0.1g g / L.

[0046] The feeding medium used in the present invention includes: glucose concentration 50%, dosage 8%; glycerin concentration 15%, dosage 4%.

[0047] (1) Take by weighing 100L of components according to the above formula;

[0048] (2) Dissolve in a specified amount of purified aqueous solution;

[0049] (3) adjust the pH value to 5.0-8.0 with sodium hydroxide solution;

[0050] (4) The fermentation medium is sterilized in a fermenter of corresponding volume (121°C, 30 minutes), and the feed medium is sterilized in a corresponding special glass feeding bottle (115°C, 30 minutes); ...

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Abstract

The invention provides a high-density fermentation method of recombinant human interferon α2b, which controls the feed feeding of nutrients through online detection of glucose concentration, maintains the respiratory quotient between 0.9 and 1, and controls relatively constant dissolved oxygen, so as to achieve The purpose of maintaining the low level of organic acid is to realize the high-density fermentation of recombinant human interferon α2b.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a high-density fermentation method of recombinant human interferon α2b. Background technique [0002] The current level of high-density fermentation in my country is far behind that of foreign countries. The concentration of bacteria after fermentation and the output of products are very different from the international level. For example, the fermentation of recombinant E. coli producing recombinant human growth hormone in China can reach The OD600 of the liquid is 50-60, while abroad can reach the level of 120. The output of the target product is only 0.4g / L in China, while the foreign country can reach the level of 1.1g / L. The high-density fermentation of Pichia pastoris has more advantages. It can grow to an OD value above 500 in a synthetic medium, and the volume of the bacteria can account for more than half of the volume of the fermentation broth. [0003] In terms of high-densi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/02C12R1/19
Inventor 张春丽周德胜吉春
Owner BEIJING KAWIN TECH SHARE HLDG
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