Resolving molecular diffusion and aggregation of antibody proteins with megahertz X-ray free-electron laser pulses

Mario Reiser (), Anita Girelli, Anastasia Ragulskaya, Sudipta Das, Sharon Berkowicz, Maddalena Bin, Marjorie Ladd-Parada, Mariia Filianina, Hanna-Friederike Poggemann, Nafisa Begam, Mohammad Sayed Akhundzadeh, Sonja Timmermann, Lisa Randolph, Yuriy Chushkin, Tilo Seydel, Ulrike Boesenberg, Jörg Hallmann, Johannes Möller, Angel Rodriguez-Fernandez, Robert Rosca, Robert Schaffer, Markus Scholz, Roman Shayduk, Alexey Zozulya, Anders Madsen, Frank Schreiber, Fajun Zhang, Fivos Perakis () and Christian Gutt ()
Additional contact information
Mario Reiser: AlbaNova University Center, Stockholm University
Anita Girelli: Institut für Angewandte Physik, Universität Tübingen
Anastasia Ragulskaya: Institut für Angewandte Physik, Universität Tübingen
Sudipta Das: AlbaNova University Center, Stockholm University
Sharon Berkowicz: AlbaNova University Center, Stockholm University
Maddalena Bin: AlbaNova University Center, Stockholm University
Marjorie Ladd-Parada: AlbaNova University Center, Stockholm University
Mariia Filianina: AlbaNova University Center, Stockholm University
Hanna-Friederike Poggemann: AlbaNova University Center, Stockholm University
Nafisa Begam: Institut für Angewandte Physik, Universität Tübingen
Mohammad Sayed Akhundzadeh: Universität Siegen
Sonja Timmermann: Universität Siegen
Lisa Randolph: Universität Siegen
Yuriy Chushkin: ESRF - The European Synchrotron
Tilo Seydel: Institut Laue-Langevin
Ulrike Boesenberg: European X-Ray Free-Electron Laser Facility
Jörg Hallmann: European X-Ray Free-Electron Laser Facility
Johannes Möller: European X-Ray Free-Electron Laser Facility
Angel Rodriguez-Fernandez: European X-Ray Free-Electron Laser Facility
Robert Rosca: European X-Ray Free-Electron Laser Facility
Robert Schaffer: European X-Ray Free-Electron Laser Facility
Markus Scholz: European X-Ray Free-Electron Laser Facility
Roman Shayduk: European X-Ray Free-Electron Laser Facility
Alexey Zozulya: European X-Ray Free-Electron Laser Facility
Anders Madsen: European X-Ray Free-Electron Laser Facility
Frank Schreiber: Institut für Angewandte Physik, Universität Tübingen
Fajun Zhang: Institut für Angewandte Physik, Universität Tübingen
Fivos Perakis: AlbaNova University Center, Stockholm University
Christian Gutt: Universität Siegen

Nature Communications, 2022, vol. 13, issue 1, 1-10

Abstract: Abstract X-ray free-electron lasers (XFELs) with megahertz repetition rate can provide novel insights into structural dynamics of biological macromolecule solutions. However, very high dose rates can lead to beam-induced dynamics and structural changes due to radiation damage. Here, we probe the dynamics of dense antibody protein (Ig-PEG) solutions using megahertz X-ray photon correlation spectroscopy (MHz-XPCS) at the European XFEL. By varying the total dose and dose rate, we identify a regime for measuring the motion of proteins in their first coordination shell, quantify XFEL-induced effects such as driven motion, and map out the extent of agglomeration dynamics. The results indicate that for average dose rates below 1.06 kGy μs−1 in a time window up to 10 μs, it is possible to capture the protein dynamics before the onset of beam induced aggregation. We refer to this approach as correlation before aggregation and demonstrate that MHz-XPCS bridges an important spatio-temporal gap in measurement techniques for biological samples.

Date: 2022
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DOI: 10.1038/s41467-022-33154-7

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