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Analytical band centrifugation revisited
Cornelia M Schneider1, Helmut Cölfen2
1Physical Chemistry, University of Konstanz, Universitätsstrasse 10, 78457, Constance, Germany.
European Biophysics Journal : EBJ
|June 23, 2018
Summary
Analytical band centrifugation (ABC) offers advantages for macromolecule and nanoparticle analysis. Accounting for diffusion and dynamic density gradients enhances data evaluation, enabling sophisticated analysis beyond average sedimentation coefficients.
Area of Science:
- Biophysical characterization
- Analytical ultracentrifugation
- Macromolecular and nanoparticle analysis
Background:
- Analytical band centrifugation (ABC) is a powerful technique for analyzing macromolecules and nanoparticles.
- Current ABC data analysis is restricted, limiting its potential compared to sedimentation velocity (SV) experiments.
- ABC allows physical separation and in-situ chemical reactions, offering unique advantages.
Purpose of the Study:
- To investigate the integration of ABC data for analysis using established SV evaluation programs.
- To identify and address limiting issues in ABC data analysis.
- To enhance the sophistication of ABC data interpretation.
Main Methods:
- Integration of ABC data with established sedimentation velocity (SV) analysis software.
- Testing with myoglobin (high diffusion) and 100 nm polystyrene latex particles (low diffusion).
- Modeling the influence of boundary broadening due to diffusion and dynamic density gradients.
Main Results:
- Identified boundary broadening by diffusion and dynamic density gradients as key limitations.
- Demonstrated that diffusion effects can be accounted for in the analysis.
- Showed the possibility of calculating time-dependent solvent density and viscosity changes in the AUC cell.
Conclusions:
- Accounting for the dynamic density gradient is crucial for advanced ABC data analysis.
- This approach will enable sophisticated methods analyzing the entire sedimentation boundary.
- The findings will expand ABC applications beyond determining average sedimentation coefficients.
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