Related Experiment Videos
CO2 and fluorinated solvent-based technologies for protein microparticle precipitation from aqueous solutions
Marazban Sarkari1, Inaas Darrat, Barbara L Knutson
1Department of Chemical and Materials Engineering, University of Kentucky, Lexington, Kentucky 40506, USA.
Biotechnology Progress
|April 5, 2003
Summary
Researchers developed a new method for protein microparticle precipitation using ethanol as a cosolvent. This technique improves the solubility of proteins in compressed carbon dioxide (CO2) or hydrofluoroether (HFE) antisolvents, preserving protein activity.
Area of Science:
- Biotechnology
- Chemical Engineering
- Materials Science
Background:
- Compressed or supercritical fluid antisolvent (PCA) precipitation is used for microparticle production.
- Limited protein solubility in traditional PCA systems (water/CO2) restricts applications.
- Ethanol as a cosolvent enhances PCA for aqueous protein solutions.
Purpose of the Study:
- To extend PCA to a wider range of proteins in aqueous solutions.
- To utilize ethanol as a cosolvent to improve CO2 antisolvent properties.
- To precipitate alpha-chymotrypsin using compressed CO2 and a hydrofluoroether (HFE) antisolvent.
Main Methods:
- Examined equilibrium phase behavior of antisolvent/ethanol/water systems.
- Identified one-phase regions for protein precipitation.
- Precipitated alpha-chymotrypsin using compressed CO2 and HFE antisolvents.
Main Results:
- Spherical protein microparticles (0.2-0.6 microm) were successfully recovered.
- Both CO2 and HFE antisolvents yielded microparticles with significant protein activity.
- HFE-precipitated chymotrypsin exhibited higher retained activity compared to CO2-precipitated protein.
Conclusions:
- Ethanol effectively enhances PCA for aqueous protein precipitation.
- PCA with CO2 and HFE antisolvents can produce active protein microparticles.
- HFE may be a preferred antisolvent for preserving higher protein activity.