AF4-to-SAXS: expanded characterization of nanoparticles and proteins at the P12 BioSAXS beamline

AF4-to-SAXS: expanded characterization of nanoparticles and proteins at the P12 BioSAXS beamline

Biological small-angle X-ray scattering (SAXS) is a versatile and powerful technique for investigating the structural and biophysical properties of biologically and pharmaceutically relevant macromolecules and nanoparticles. SAXS offers detailed insights into macromolecular composition, size, shape...

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Main Authors: Stefano Da Vela, Kim Bartels, Daniel Franke, Dymtro Soloviov, Tobias Gräwert, Dmitry Molodenskiy, Bastian Kolb, Christoph Wilhelmy, Roland Drexel, Florian Meier, Heinrich Haas, Peter Langguth, Melissa A. Graewert
Format: Article
Language:English
Published: International Union of Crystallography 2025-07-01
Series:Journal of Synchrotron Radiation
Subjects:
af4
biological saxs
p12 beamline
proteins
nanoparticles
full automation
operator-friendly guidelines
Online Access:https://journals.iucr.org/paper?S1600577525003959
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Summary:Biological small-angle X-ray scattering (SAXS) is a versatile and powerful technique for investigating the structural and biophysical properties of biologically and pharmaceutically relevant macromolecules and nanoparticles. SAXS offers detailed insights into macromolecular composition, size, shape and internal structure, while addressing key aspects such as oligomeric state, stability, molecular interactions, and conformational flexibility. Recently, asymmetrical-flow field-flow fractionation (AF4) was successfully coupled to SAXS, enabling online size-based fractionation and analysis of polydisperse samples. This approach allows precise, size-dependent characterization, offering significant advancements in the study of polydisperse systems. We have integrated an AF4 device at the P12 beamline at the European Molecular Biology Laboratory and implemented technical adaptations allowing full automation to make the system suitable for routine user access. We provide streamlined workflows and troubleshooting resources for both novice and advanced SAXS users thereby equipping them with clear guidance on performing AF4–SAXS measurements. The general principles of our set-up are easily adaptable to other beamlines which have integrated (or are planning to integrate) a similar system.
ISSN:1600-5775

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