In situ microfluidic dialysis for biological small-angle X-ray scattering

Magda Skou, Soren Skou, Thomas Glasdam Jensen, Bente Vestergaard, Richard E. Gillilan

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    Abstract

    Owing to the demand for low sample consumption and automated sample changing capabilities at synchrotron small-angle X-ray (solution) scattering (SAXS) beamlines, X-ray microfluidics is receiving continuously increasing attention. Here, a remote-controlled microfluidic device is presented for simultaneous SAXS and ultraviolet absorption measurements during protein dialysis, integrated directly on a SAXS beamline. Microfluidic dialysis can be used for monitoring structural changes in response to buffer exchange or, as demonstrated, protein concentration. By collecting X-ray data during the concentration procedure, the risk of inducing protein aggregation due to excessive concentration and storage is eliminated, resulting in reduced sample consumption and improved data quality. The proof of concept demonstrates the effect of halted or continuous flow in the microfluidic device. No sample aggregation was induced by the concentration process at the levels achieved in these experiments. Simulations of fluid dynamics and transport properties within the device strongly suggest that aggregates, and possibly even higher-order oligomers, are preferentially retained by the device, resulting in incidental sample purification. Hence, this versatile microfluidic device enables investigation of experimentally induced structural changes under dynamically controllable sample conditions. (C) 2014 International Union of Crystallography
    Original languageEnglish
    JournalJournal of Applied Crystallography
    Volume47
    Issue number4
    Pages (from-to)1355-1366
    Number of pages12
    ISSN0021-8898
    DOIs
    Publication statusPublished - 2014

    Bibliographical note

    © 2014 International Union of Crystallography

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