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Column chromatographic separation of cells using aqueous polymeric two-phase systems.
D R Skuse1, W Müller, D E Brooks
1Department of Chemistry, University of British Columbia, Vancouver, Canada.
Analytical Biochemistry
|November 1, 1988
Summary
This study introduces a novel chromatography method for cell separation using immobilized dextran/polyethylene glycol (PEG) phases. This technique efficiently separates mammalian erythrocytes based on cell membrane interactions.
Area of Science:
- Biochemistry
- Separation Science
- Cell Biology
Background:
- Aqueous polymeric two-phase systems are established for cell separation.
- Multistep countercurrent distribution is often required for cells with similar partition coefficients, but faces time and apparatus limitations.
Purpose of the Study:
- To develop an alternative chromatographic technique for cell separation.
- To overcome the limitations of traditional countercurrent distribution methods.
Main Methods:
- Immobilization of the dextran-rich phase of a dextran/polyethylene glycol (PEG) system onto derivatized agarose beads.
- Utilizing the PEG-rich phase as the eluent.
- Incorporating PEG-fatty acid affinity ligand gradients into the eluent for separation.
Main Results:
- Developed a chromatographic technique for cell separation.
- Achieved separation of mammalian erythrocytes based on differential fatty acid-membrane interactions.
- Successfully demonstrated a model separation of dog and human erythrocytes.
Conclusions:
- The developed chromatographic method offers a viable alternative to traditional cell separation techniques.
- Affinity ligand gradients in the eluent enable specific cell separations based on membrane properties.