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Online ion‐exchange chromatography for small‐angle X‐ray scattering
Author(s) -
Hutin Stephanie,
Brennich Martha,
Maillot Benoit,
Round Adam
Publication year - 2016
Publication title -
acta crystallographica section d
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.374
H-Index - 138
ISSN - 2059-7983
DOI - 10.1107/s2059798316012833
Subject(s) - scattering , ion chromatography , dispersity , small angle scattering , materials science , beamline , particle size , size exclusion chromatography , background subtraction , analytical chemistry (journal) , optics , chromatography , chemistry , physics , nuclear magnetic resonance , pixel , polymer chemistry , enzyme , beam (structure)
Biological small‐angle X‐ray scattering (BioSAXS) is a powerful technique to determine the solution structure, particle size, shape and surface‐to‐volume ratio of macromolecules. However, a drawback is that the sample needs to be monodisperse. To ensure this, size‐exclusion chromatography (SEC) has been implemented on many BioSAXS beamlines. Here, the integration of ion‐exchange chromatography (IEC) using both continuous linear and step gradients on a beamline is described. Background subtraction for continuous gradients by shifting a reference measurement and two different approaches for step gradients, which are based on interpolating between two background measurements, are discussed. The results presented here serve as a proof of principle for online IEC and subsequent data treatment.