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Updated: Jun 5, 2026
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Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain
Published on: December 12, 2017
[Refolding of reduced/denatured RNase A the hydrophobic liquid-solid interface]
1Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Institute of Modern Separation Sciences, Key Laboratory of Separation Science in Shaanxi Province, Northwest University, Xi'an 710069, China.
Se Pu = Chinese Journal of Chromatography
|January 26, 2011
Summary
Researchers optimized conditions for refolding denatured Ribonuclease A (RNase A) using hydrophobic interaction chromatography (HIC). Optimal refolding was achieved at pH 8.0 with 2.0 mol/L urea and an 8:1 GSH/GSSG ratio, demonstrating HIC
Area of Science:
- Biochemistry and Biophysics
- Protein Chemistry
- Chromatography Techniques
Context:
- Ribonuclease A (RNase A) is a crucial enzyme in molecular biology.
- Protein denaturation can lead to loss of enzymatic activity.
- Renaturation techniques are vital for restoring protein function.
Purpose:
- To investigate the renaturation of reduced/denatured RNase A.
- To explore the effectiveness of hydrophobic interaction chromatography (HIC) for protein refolding.
- To determine optimal conditions for RNase A refolding, including urea concentration, glutathione redox buffer (GSH/GSSG), pH, and protein concentration.
Summary:
- Hydrophobic interaction chromatography (HIC) was employed to study RNase A renaturation.
- Optimal refolding conditions were identified as pH 8.0, 2.0 mol/L urea, and an 8:1 ratio of reduced to oxidized glutathione (GSH/GSSG).
- High bioactivity efficiency (98.0%) and mass recovery (61.9%) were achieved for urea-denatured RNase A, and even higher for guanidine hydrochloride-denatured RNase A (100.1% and 66.8%, respectively).
Impact:
- HIC proves to be a powerful and novel approach for protein refolding.
- The study provides optimized protocols for restoring RNase A activity.
- This research contributes to advancements in protein purification and renaturation methodologies.
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