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Protein partitioning in detergent-based aqueous two-phase systems
G C Terstappen1, R A Ramelmeier, M R Kula
1Institut für Enzymtechnologie, Heinrich-Heine-Universität Düsseldorf, Forschungszentrum Jülich, Germany.
Journal of Biotechnology
|April 1, 1993
Summary
Nonionic polyoxyethylene detergents form two liquid phases above their cloud point. Protein partitioning into the detergent-rich phase increases with protein hydrophobicity, indicating hydrophobic interactions drive this separation.
Area of Science:
- Biochemistry
- Physical Chemistry
- Separation Science
Background:
- Aqueous solutions of nonionic polyoxyethylene detergents exhibit phase separation into two liquid phases above a specific temperature, known as the cloud point.
- One phase is detergent-enriched (coacervate phase), while the other is detergent-depleted.
- These systems offer potential for protein separation and purification.
Purpose of the Study:
- To systematically and quantitatively investigate protein partitioning in detergent-based aqueous two-phase systems.
- To elucidate the role of protein hydrophobicity in partitioning behavior.
- To explore the applicability of these systems for various protein types.
Main Methods:
- Utilized a series of similar polyoxyethylene detergents with varying alkyl chain lengths.
- Employed proteins with diverse hydrophobicity levels.
- Analyzed protein partitioning quantitatively across the two liquid phases.
Main Results:
- Increasing detergent alkyl chain length, temperature, or salt concentration enhanced detergent separation into the coacervate phase.
- A concomitant increase in protein partitioning into the coacervate phase was observed.
- A positive correlation was found between protein hydrophobicity and its partitioning into the coacervate phase.
Conclusions:
- Protein-detergent interactions in these systems are primarily driven by hydrophobic forces.
- The study confirms the utility of detergent-based aqueous two-phase systems for separating proteins.
- These systems are applicable to both membrane proteins and water-soluble proteins with hydrophobic domains.