Quantitative size-resolved characterization of mRNA nanoparticles by in-line coupling of asymmetrical-flow field-flow fractionation with small angle X-ray scattering

  • Sci Rep. 2023 Sep 22;13(1):15764. doi: 10.1038/s41598-023-42274-z.
Melissa A Graewert  #  1 ,  Christoph Wilhelmy  #  2 ,  Tijana Bacic  #  3 ,  Jens Schumacher  3 ,  Clement Blanchet  4 ,  Florian Meier  5 ,  Roland Drexel  5 ,  Roland Welz  5 ,  Bastian Kolb  2 ,  Kim Bartels  2 ,  Thomas Nawroth  2 ,  Thorsten Klein  5 ,  Dmitri Svergun  4  6 ,  Peter Langguth  2 ,  Heinrich Haas  7  8
Affiliations
  • 1. European Molecular Biology Laboratory, Hamburg Unit, Hamburg, Germany. [email protected].
  • 2. Department of Biopharmaceutics and Pharmaceutical Technology, Johannes Gutenberg-University, Mainz, Germany.
  • 3. BioNTech SE, Mainz, Germany.
  • 4. European Molecular Biology Laboratory, Hamburg Unit, Hamburg, Germany.
  • 5. Postnova Analytics GmbH, Landsberg am Lech, Germany.
  • 6. BIOSAXS GmbH, Hamburg, Germany.
  • 7. Department of Biopharmaceutics and Pharmaceutical Technology, Johannes Gutenberg-University, Mainz, Germany. [email protected].
  • 8. BioNTech SE, Mainz, Germany. [email protected].
  • # Contributed equally.
PMID: 37737457 DOI: 10.1038/s41598-023-42274-z
Abstract

We present a generically applicable approach to determine an extensive set of size-dependent critical quality attributes inside nanoparticulate pharmaceutical products. By coupling asymmetrical-flow field-flow fractionation (AF4) measurements directly in-line with solution small angle X-ray scattering (SAXS), vital information such as (i) quantitative, absolute size distribution profiles, (ii) drug loading, (iii) size-dependent internal structures, and (iv) quantitative information on free drug is obtained. Here the validity of the method was demonstrated by characterizing complex mRNA-based lipid nanoparticle products. The approach is particularly applicable to particles in the size range of 100 nm and below, which is highly relevant for pharmaceutical products-both biologics and nanoparticles. The method can be applied as well in Other fields, including structural biology and environmental sciences.

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