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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

Bolinsson, Hans LU ; Söderberg, Christopher ; Herranz-Trillo, Fátima ; Wahlgren, Marie LU orcid and Nilsson, Lars LU (2023) In Analytical and Bioanalytical Chemistry 415(25). p.6237-6246
Abstract

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... (More)

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.]

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author
; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
AF4, AF4-UV-SAXS on-line coupling, Monoclonal antibody, Protein, SAXS
in
Analytical and Bioanalytical Chemistry
volume
415
issue
25
pages
10 pages
publisher
Springer
external identifiers
  • pmid:37572213
  • scopus:85167804655
ISSN
1618-2642
DOI
10.1007/s00216-023-04900-7
language
English
LU publication?
yes
id
77229508-a4c0-4add-a0e0-034d19b2b7ec
date added to LUP
2023-12-01 13:27:58
date last changed
2024-04-14 13:51:00
@article{77229508-a4c0-4add-a0e0-034d19b2b7ec,
  abstract     = {{<p>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.]</p>}},
  author       = {{Bolinsson, Hans and Söderberg, Christopher and Herranz-Trillo, Fátima and Wahlgren, Marie and Nilsson, Lars}},
  issn         = {{1618-2642}},
  keywords     = {{AF4; AF4-UV-SAXS on-line coupling; Monoclonal antibody; Protein; SAXS}},
  language     = {{eng}},
  number       = {{25}},
  pages        = {{6237--6246}},
  publisher    = {{Springer}},
  series       = {{Analytical and Bioanalytical Chemistry}},
  title        = {{Realizing the AF4-UV-SAXS on-line coupling on protein and antibodies using high flux synchrotron radiation at the CoSAXS beamline, MAX IV}},
  url          = {{http://dx.doi.org/10.1007/s00216-023-04900-7}},
  doi          = {{10.1007/s00216-023-04900-7}},
  volume       = {{415}},
  year         = {{2023}},
}