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Method for highly efficient renaturation of membrane albumen

A membrane protein, protein technology, applied in the renaturation of proteins, renaturation, renaturation and purification of recombinant membrane protein inclusion bodies expressed in prokaryotic cells, and protein purification. Small, high purity, and the effect of improving the protein renaturation rate

Active Publication Date: 2007-10-17
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Abstract
  • Description
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Problems solved by technology

[0003] However, the renaturation of proteins has always been a difficult and hot spot in bioengineering
And because of its strong hydrophobicity, the difficulty of renaturation is much more difficult than that of ordinary proteins.

Method used

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  • Method for highly efficient renaturation of membrane albumen
  • Method for highly efficient renaturation of membrane albumen
  • Method for highly efficient renaturation of membrane albumen

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Embodiment Construction

[0034] Combining the long-term research on the properties of membrane proteins and the characteristics of various chromatographic renaturation methods, the applicant invented a simple, fast and efficient method for renaturation and purification of membrane proteins, and obtained high-purity, high-uniformity, high-activity The product protein can be widely used in various research (such as protein structure research, etc.), and this method has the potential of large-scale amplification.

[0035] The method of the present invention uses hydrophobic chromatography and ion exchange combination system to renature and purify proteins, and is characterized in that the method firstly purifies the dissolved inclusion body protein in a hydrophobic chromatography column, then uses molecular sieves to desalt, and finally Denaturing agent concentration elution and salt concentration gradient elution are carried out in ion exchange chromatography to induce protein to fold and refold correctl...

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Abstract

The invention discloses a high effectively membrane protein renaturation method, which comprise: purifying dissolved inclusion bodies protein in hydrophobic chromatography column, desalting using molecular sieve, carrying out denaturant concentration gradient elution and salt concentration gradient elution through ion exchange column chromatography, thus induction protein gradually folding right and renaturing . Any small molecule additive (as GSSG,GSH, PDI molecular chaperon etc.) is not employed in the complete process which reduce the cost. High uniformity, high activity target protein of above 98% in purity is obtained through the method which can be conveniently and continuously operated. The cost of the method is low.

Description

technical field [0001] The invention belongs to the protein purification and renaturation technology in the technical field of bioengineering, and mainly relates to a method for renaturation and purification of inclusion bodies of recombinant membrane proteins expressed by prokaryotic cells, especially a method for renaturation of proteins containing Ig domain. Background technique [0002] The current rapid development of biotechnology has made it possible to produce target proteins on a large scale. Some proteins that are rare and difficult to extract can be highly expressed in host cells through gene recombination technology, and the most commonly used host cells are Escherichia coli. The expression of target protein in Escherichia coli has the advantages of rapidity, large quantity and low cost. However, the massive expression of foreign proteins in Escherichia coli often forms protein aggregates, that is, inclusion bodies. Inclusion bodies are a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14
Inventor 陈志南宋斐唐浩
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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