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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| Format: | Article |
| Language: | English |
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International Union of Crystallography
2025-07-01
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| Series: | Journal of Synchrotron Radiation |
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| Online Access: | https://journals.iucr.org/paper?S1600577525003959 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Stefano Da Vela |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-b5ebd5bd5ba345d986a70a104e0c462d |
| institution | Kabale University |
| issn | 1600-5775 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | International Union of Crystallography |
| record_format | Article |
| series | Journal of Synchrotron Radiation |
| spelling | doaj-art-b5ebd5bd5ba345d986a70a104e0c462d2025-08-20T03:33:13ZengInternational Union of CrystallographyJournal of Synchrotron Radiation1600-57752025-07-0132497198510.1107/S1600577525003959ju5081AF4-to-SAXS: expanded characterization of nanoparticles and proteins at the P12 BioSAXS beamlineStefano Da Vela0Kim Bartels1Daniel Franke2Dymtro Soloviov3Tobias Gräwert4Dmitry Molodenskiy5Bastian Kolb6Christoph Wilhelmy7Roland Drexel8Florian Meier9Heinrich Haas10Peter Langguth11Melissa A. Graewert12European Molecular Biology Laboratory, Hamburg Unit, Hamburg, GermanyDepartment of Biopharmaceutics and Pharmaceutical Technology, Johannes Gutenberg University, Mainz, GermanyBIOSAXS GmbH, Hamburg, GermanyEuropean Molecular Biology Laboratory, Hamburg Unit, Hamburg, GermanyBIOSAXS GmbH, Hamburg, GermanyEuropean Molecular Biology Laboratory, Hamburg Unit, Hamburg, GermanyDepartment of Biopharmaceutics and Pharmaceutical Technology, Johannes Gutenberg University, Mainz, GermanyDepartment of Biopharmaceutics and Pharmaceutical Technology, Johannes Gutenberg University, Mainz, GermanyPostnova Analytics GmbH, Landsberg am Lech, GermanyPostnova Analytics GmbH, Landsberg am Lech, GermanyDepartment of Biopharmaceutics and Pharmaceutical Technology, Johannes Gutenberg University, Mainz, GermanyDepartment of Biopharmaceutics and Pharmaceutical Technology, Johannes Gutenberg University, Mainz, GermanyEuropean Molecular Biology Laboratory, Hamburg Unit, Hamburg, GermanyBiological 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.https://journals.iucr.org/paper?S1600577525003959af4biological saxsp12 beamlineproteinsnanoparticlesfull automationoperator-friendly guidelines |
| spellingShingle | 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 AF4-to-SAXS: expanded characterization of nanoparticles and proteins at the P12 BioSAXS beamline Journal of Synchrotron Radiation af4 biological saxs p12 beamline proteins nanoparticles full automation operator-friendly guidelines |
| title | AF4-to-SAXS: expanded characterization of nanoparticles and proteins at the P12 BioSAXS beamline |
| title_full | AF4-to-SAXS: expanded characterization of nanoparticles and proteins at the P12 BioSAXS beamline |
| title_fullStr | AF4-to-SAXS: expanded characterization of nanoparticles and proteins at the P12 BioSAXS beamline |
| title_full_unstemmed | AF4-to-SAXS: expanded characterization of nanoparticles and proteins at the P12 BioSAXS beamline |
| title_short | AF4-to-SAXS: expanded characterization of nanoparticles and proteins at the P12 BioSAXS beamline |
| title_sort | af4 to saxs expanded characterization of nanoparticles and proteins at the p12 biosaxs beamline |
| topic | af4 biological saxs p12 beamline proteins nanoparticles full automation operator-friendly guidelines |
| url | https://journals.iucr.org/paper?S1600577525003959 |
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