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Purifying method for recombination human leukopenic-1 receptor antagonist

A technology of interleukin and receptor antagonist, which is applied in the field of protein purification technology, can solve the problems of long process cycle, low product recovery rate, high production cost, etc., and achieve good reproducibility, simplified operation process, good The effect of technical support

Inactive Publication Date: 2006-09-06
无锡大周生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The purification process is complicated, the process cycle is long, the process requirements are high (generally need to be carried out in a low temperature environment), and the product recovery rate is low
High production costs

Method used

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Examples

Experimental program
Comparison scheme
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Embodiment Construction

[0022] Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods without specific conditions indicated in the following examples are usually operated under conventional conditions well known in the art.

[0023] 1. Purification process

[0024] (1) Fermentation

[0025] Medium: 2×YT

[0026] Tryptone 10.0g

[0027] Yeast Extract 10.0g

[0028] NaCl 5.0g

[0029] Add deionized water to 1000ml

[0030] Process parameters:

[0031] Growth temperature: 30°C

[0032] Induction temperature: 42°C

[0033] pH: 7.2

[0034] Dissolved Oxygen: 45%

[0035] Fermentation process route: preserved strain - single colony - first-class seed (F 2 )---secondary seed (F 3 )Cultivation--fermentation--centrifugation to collect the bacteria.

[0036] Activation and expansio...

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PUM

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Abstract

A process for purifying the recombinant human interleukin-1 receptor antagon features use of 2-step liquid-phase chromatography method including anionic exchange liquid-phase chromatography and cationic exchange chromatography. Its advantages are high purity and high activity.

Description

field of invention [0001] The invention relates to a protein purification process, in particular to a purification process capable of large-scale preparation of protein cytokine receptor antagonists. Background of the invention [0002] Recombinant interleukin-1 receptor antagonist (rhIL-1ra) is a protein cytokine receptor antagonist existing in the human body, which specifically blocks the activity of IL-1 by binding to interleukin-1 receptor. In 1990, Eisenberg et al. from Synergen Corporation in the United States reported the full-length cDNA sequence for the first time in the British journal Nature (Nature 1990; 343: 341-346). [0003] IL-1 plays a key role in initiating the inflammatory response of the body. IL-1 is involved in the occurrence and development of many human diseases. IL-1 itself can not only attract inflammatory response cells in the body to the site of inflammation, but also stimulate the production of various inflammatory mediators in the body. This f...

Claims

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Application Information

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IPC IPC(8): C07K14/435C07K1/18C07K1/16C07K14/545
Inventor 马大龙狄春辉朱仁英杨璐韩普
Owner 无锡大周生物工程有限公司