title: Online ion-exchange chromatography for small-angle X-ray scattering.
abstract: 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.
authors.affiliation: Institut de Biologie Structurale, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Hutin
authors.affiliation: European Synchrotron Radiation Facilty, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Martha
authors.initials: M
authors.lastname: Brennich
authors.affiliation: European Synchrotron Radiation Facilty, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Maillot
authors.affiliation: EMBL Grenoble Outstation, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Adam
authors.initials: A
authors.lastname: Round
mesh_terms: D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D053838:Scattering, Small Angle; D012710:Serum Albumin, Bovine / Q000737:chemistry; D014616:Vaccinia virus / Q000737:chemistry; D014764:Viral Proteins / Q000737:chemistry; D014961:X-Ray Diffraction / Q000379:methods*
chemical_list: D011506:Proteins; D014764:Viral Proteins; D012710:Serum Albumin, Bovine
keywords: BioSAXS; ion-exchange chromatography; online purification