Small-angle X-ray Scattering (SAXS) has been a versatile technique for studying biomolecules in solution for several decades now. Developments in SAXS techniques that integrate in situ purification with a high-throughput, multimodal design philosophy have revolutionized the reach and tempo of BioSAXS experiments. The current zenith of the field comes in the form of size exclusion chromatography coupled SAXS with in-line multiangle light scattering (SEC-SAXS-MALS). This technique has been a considerable focus at the Structurally Integrated BiologY for Life Sciences (SIBYLS) beamline at the Advanced Light Source (ALS) in Berkeley, California, over the last 5 years and continues to be a point of active development. In this chapter, we describe the design of the SEC-SAXS-MALS system and general guidelines for collecting, processing, and analyzing SEC-SAXS-MALS data at the SIBYLS beamline.
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Size exclusion chromatography coupled small angle X-ray scattering with tandem multiangle light scattering at the SIBYLS beamline.
Methods Enzymol
November 2022
Molecular Biophysics and Integrated Bioimaging, Lawrence Berkeley National Laboratory, Berkeley, CA, United States. Electronic address:
Small-angle X-ray Scattering (SAXS) has been a versatile technique for studying biomolecules in solution for several decades now. Developments in SAXS techniques that integrate in situ purification with a high-throughput, multimodal design philosophy have revolutionized the reach and tempo of BioSAXS experiments. The current zenith of the field comes in the form of size exclusion chromatography coupled SAXS with in-line multiangle light scattering (SEC-SAXS-MALS).
View Article and Find Full Text PDFHigh-throughput SAXS for the characterization of biomolecules in solution: a practical approach.
Methods Mol Biol
June 2014
Physcial Bioscience Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
The recent innovation of collecting X-ray scattering from solutions containing purified macromolecules in high-throughput has yet to be truly exploited by the biological community. Yet, this capability is becoming critical given that the growth of sequence and genomics data is significantly outpacing structural biology results. Given the huge mismatch in information growth rates between sequence and structural methods, their combined high-throughput and high success rate make high-throughput small angle X-ray scattering (HT-SAXS) analyses increasingly valuable.
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J Appl Crystallogr
February 2013
Physical Bioscience Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
The SIBYLS beamline (12.3.1) of the Advanced Light Source at Lawrence Berkeley National Laboratory, supported by the US Department of Energy and the National Institutes of Health, is optimized for both small-angle X-ray scattering (SAXS) and macromolecular crystallography (MX), making it unique among the world's mostly SAXS or MX dedicated beamlines.
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Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
Biological small-angle X-ray scattering (SAXS) provides powerful complementary data for macromolecular crystallography (MX) by defining shape, conformation and assembly in solution. Although SAXS is in principle the highest throughput technique for structural biology, data collection is limited in practice by current data collection software. Here the adaption of beamline control software, historically developed for MX beamlines, for the efficient operation and high-throughput data collection at synchrotron SAXS beamlines is reported.
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