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Realizing the AF4-UV-SAXS on-line coupling on protein and antibodies using high flux synchrotron radiation at the CoSAXS beamline, MAX IV
Lund University, Sweden.
RISE Research Institutes of Sweden, Bioeconomy and Health, Chemical Process and Pharmaceutical Development.
MAX IV Laboratory, Sweden.
Lund University, Sweden.
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2023 (English)In: Analytical and Bioanalytical Chemistry, ISSN 1618-2642, E-ISSN 1618-2650, Vol. 415, p. 6237-Article in journal (Refereed) Published
Abstract [en]

In this paper, we demonstrate the coupling of synchrotron small angle X-ray scattering (SAXS) to asymmetrical flow-field flow fractionation (AF4) for protein characterization. To the best of our knowledge, this is the first time AF4 is successfully coupled to a synchrotron for on-line measurements on proteins. This coupling has potentially high impact, as it opens the possibility to characterize individual constituents of sensitive and/or complex samples, not suited for separation using other techniques, and for low electron density samples where high X-ray flux is required, e.g., biomolecules and biologics. AF4 fractionates complex samples in native or close to native environment, with low shear forces and system surface area. Many orders of magnitude in size can be fractionated in one measurement, without having to reconfigure the experimental setup. We report AF4 fractionations with correlated UV and statistically adequate SAXS data of bovine serum albumin and a monoclonal antibody and evaluate SAXS data recorded for the two protein systems. Graphical Abstract: [Figure not available: see fulltext.] © 2023, The Author(s).

Place, publisher, year, edition, pages
Springer Science and Business Media Deutschland GmbH , 2023. Vol. 415, p. 6237-
Keywords [en]
AF4, AF4-UV-SAXS on-line coupling, Monoclonal antibody, Protein, SAXS, Fractionation, Liquid chromatography, Mammals, Monoclonal antibodies, Synchrotron radiation, Synchrotrons, AF4-UV-small angle X-ray scattering on-line coupling, Beam-lines, Complex samples, High flux, On-line coupling, Scattering data, Small angle X-ray scattering, Synchrotron small angle X-ray scatterings, X ray scattering
National Category
Engineering and Technology
Identifiers
URN: urn:nbn:se:ri:diva-66125DOI: 10.1007/s00216-023-04900-7Scopus ID: 2-s2.0-85167804655OAI: oai:DiVA.org:ri-66125DiVA, id: diva2:1796108
Note

Research conducted at MAX IV, a Swedish national user facility, is supported by the Swedish Research council under contract 2018-07152, the Swedish Governmental Agency for Innovation Systems under contract 2018-04969, and Formas under contract 2019-02496.

Available from: 2023-09-11 Created: 2023-09-11 Last updated: 2024-05-27Bibliographically approved

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