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Crystallization of Proteins on Chip by Microdialysis for In Situ X-ray Diffraction Studies
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In situ microfluidic dialysis for biological small-angle X-ray scattering
Magda Skou1, Søren Skou2, Thomas G Jensen3
1Department of Drug Design and Pharmacology, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark.
Journal of Applied Crystallography
|September 23, 2014
Summary
This study introduces a microfluidic device for simultaneous small-angle X-ray scattering (SAXS) and UV absorption measurements. This method reduces sample consumption and prevents protein aggregation during concentration.
Area of Science:
- Biophysics
- Materials Science
- Analytical Chemistry
Background:
- Synchrotron small-angle X-ray scattering (SAXS) requires low sample consumption and automated sample handling.
- X-ray microfluidics is gaining attention for its potential in automated and low-volume biological sample analysis.
Purpose of the Study:
- To develop and demonstrate a remote-controlled microfluidic device for simultaneous SAXS and UV absorption measurements.
- To monitor protein structural changes and concentration in real-time during dialysis on a SAXS beamline.
- To eliminate sample aggregation risk during protein concentration, thereby reducing sample consumption and improving data quality.
Main Methods:
- Integration of a microfluidic device directly onto a SAXS beamline for remote-controlled measurements.
- Simultaneous SAXS and ultraviolet absorption measurements during protein dialysis and concentration.
- Fluid dynamics and transport simulations to analyze sample behavior within the microfluidic device.
Main Results:
- The microfluidic device enabled simultaneous SAXS and UV absorption measurements during protein concentration without inducing aggregation.
- Experiments demonstrated the effect of flow (halted or continuous) on sample behavior within the device.
- Simulations indicated preferential retention of aggregates and oligomers by the device, suggesting incidental sample purification.
Conclusions:
- The developed microfluidic device facilitates investigation of protein structural changes under dynamically controlled conditions.
- This technology reduces sample consumption and enhances data quality in SAXS experiments.
- The device offers a versatile platform for real-time monitoring and potential purification of biological samples.
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