期刊
CRYSTALS
卷 10, 期 11, 页码 -出版社
MDPI
DOI: 10.3390/cryst10110975
关键词
small angle X-ray scattering; size exclusion chromatography; laser light scattering; data quality; benchmarked datasets; SASBDB
资金
- iNEXT-Discovery - Horizon 2020 program of the European Commission [871037]
- German Ministry of Science and Education [16QK10A-SAS-BSOFT]
- Biomedical Physics of Infection grant of the Joachim Herz Stiftung (Hamburg, Germany) [900000]
We describe the updated size-exclusion chromatography small angle X-ray scattering (SEC-SAXS) set-up used at the P12 bioSAXS beam line of the European Molecular Biology Laboratory (EMBL) at the PETRAIII synchrotron, DESY Hamburg (Germany). The addition of size exclusion chromatography (SEC) directly on-line to the SAXS capillary has become a well-established approach to reduce the effects of the sample heterogeneity on the SAXS measurements. The additional use of multi-angle laser light scattering (MALLS), UV absorption spectroscopy, refractive index (RI), and quasi-elastic light scattering (QELS) in parallel to the SAXS measurements enables independent molecular weight validation and hydrodynamic radius estimates. This allows one to address sample monodispersity as well as conformational heterogeneity. The benefits of the current SEC-SAXS set-up are demonstrated on a set of selected standard proteins. The processed SEC-SAXS data and models are provided in the Small Angle Scattering Biological Data Bank (SASBDB) and are labeled as bench-marked datasets that include the unsubtracted data frames spanning the respective SEC elution profiles and corresponding MALLS-UV-RI-QELS data. These entries provide method developers with datasets suitable for testing purposes, in addition to an educational resource for SAS data analysis and modeling.
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