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Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain
Published on: December 12, 2017
Urea-gradient protein refolding in size exclusion chromatography
1Institute of Modern Separation Science, College of Chemistry and Materials Science, Northwest University, 229 TaiBai North Road, Xi'an 710069, P R China. czwang@nwu.edu.cn
Current Pharmaceutical Biotechnology
|March 10, 2010
Summary
Urea gradient size exclusion chromatography (UGSEC) enhances protein refolding efficiency. This novel method optimizes operating factors for improved protein recovery, as demonstrated with recombinant human granulocyte colony stimulating factor.
Area of Science:
- Biochemistry
- Protein Chemistry
- Chromatography
Background:
- Conventional size exclusion chromatography (SEC) is a common method for protein refolding.
- Challenges remain in optimizing refolding yields and efficiency for complex proteins.
Purpose of the Study:
- To introduce and detail a novel protein refolding technique: urea gradient size exclusion chromatography (UGSEC).
- To analyze the key operating parameters influencing UGSEC performance.
- To demonstrate the application of UGSEC in refolding a specific recombinant protein.
Main Methods:
- Development and implementation of urea gradient size exclusion chromatography (UGSEC).
- Systematic investigation of operating factors, including urea gradient length and final urea concentration.
- Application of UGSEC for the refolding of recombinant human granulocyte colony stimulating factor.
Main Results:
- UGSEC integrates a urea gradient into the SEC process for enhanced protein refolding.
- Detailed discussion of critical operating parameters like gradient length and final urea concentration.
- Successful refolding of recombinant human granulocyte colony stimulating factor using the UGSEC method.
Conclusions:
- Urea gradient size exclusion chromatography (UGSEC) offers an advanced approach to protein refolding.
- Optimization of operating factors is crucial for maximizing refolding efficiency with UGSEC.
- UGSEC demonstrates practical utility in the refolding of recombinant therapeutic proteins.
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