Size-exclusion chromatography small-angle X-ray scattering of water soluble proteins on a laboratory instrument

Research output: Contribution to journalJournal articleResearchpeer-review

Documents

  • vg5091

    Final published version, 1 MB, PDF document

  • sendsupfiles

    Final published version, 8 MB, PDF document

Coupling of size-exclusion chromatography with biological solution small-angle X-ray scattering (SEC-SAXS) on dedicated synchrotron beamlines enables structural analysis of challenging samples such as labile proteins and low-affinity complexes. For this reason, the approach has gained increased popularity during the past decade. Transportation of perishable samples to synchrotrons might, however, compromise the experiments, and the limited availability of synchrotron beamtime renders iterative sample optimization tedious and lengthy. Here, the successful setup of laboratory-based SEC-SAXS is described in a proof-of-concept study. It is demonstrated that sufficient quality data can be obtained on a laboratory instrument with small sample consumption, comparable to typical synchrotron SEC-SAXS demands. UV/vis measurements directly on the SAXS exposure cell ensure accurate concentration determination, crucial for direct molecular weight determination from the scattering data. The absence of radiation damage implies that the sample can be fractionated and subjected to complementary analysis available at the home institution after SEC-SAXS. Laboratory-based SEC-SAXS opens the field for analysis of biological samples at the home institution, thus increasing productivity of biostructural research. It may further ensure that synchrotron beamtime is used primarily for the most suitable and optimized samples.
Original languageEnglish
JournalJournal of Applied Crystallography
Volume51
Issue number6
Pages (from-to)1623-1632
Number of pages11
ISSN1600-5767
DOIs
Publication statusPublished - 2018

Number of downloads are based on statistics from Google Scholar and www.ku.dk


No data available

ID: 208818708