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Investigating thermally induced aggregation of Somatropin- new insights using orthogonal techniques

Västberg, Amanda LU ; Bolinsson, Hans LU ; Leeman, Mats LU ; Nilsson, Lars LU ; Nylander, Tommy LU ; Sejwal, Kushal ; Sintorn, Ida Maria ; Lidayová, Kristina ; Sjögren, Helen LU and Wahlgren, Marie LU orcid , et al. (2023) In International Journal of Pharmaceutics 637.
Abstract

Three orthogonal techniques were used to provide new insights into thermally induced aggregation of the therapeutic protein Somatropin at pH 5.8 and 7.0. The techniques were Dynamic Light Scattering (DLS), Asymmetric Flow-Field Flow-Fractionation (AF4), and the TEM-based analysis system MiniTEM™. In addition, Differential Scanning Calorimetry (DSC) was used to study the thermal unfolding and stability. DSC and DLS were used to explain the initial aggregation process and aggregation rate at the two pH values. The results suggest that less electrostatic stabilization seems to be the main reason for the faster initial aggregation at pH 5.8, i.e., closer to the isoelectric point of Somatropin. AF4 and MiniTEM were used to investigate the... (More)

Three orthogonal techniques were used to provide new insights into thermally induced aggregation of the therapeutic protein Somatropin at pH 5.8 and 7.0. The techniques were Dynamic Light Scattering (DLS), Asymmetric Flow-Field Flow-Fractionation (AF4), and the TEM-based analysis system MiniTEM™. In addition, Differential Scanning Calorimetry (DSC) was used to study the thermal unfolding and stability. DSC and DLS were used to explain the initial aggregation process and aggregation rate at the two pH values. The results suggest that less electrostatic stabilization seems to be the main reason for the faster initial aggregation at pH 5.8, i.e., closer to the isoelectric point of Somatropin. AF4 and MiniTEM were used to investigate the aggregation pathway further. Combining the results allowed us to demonstrate Somatropin's thermal aggregation pathway at pH 7.0. The growth of the aggregates appears to follow two steps. Smaller elongated aggregates are formed in the first step, possibly initiated by partly unfolded species. In the second step, occurring during longer heating, the smaller aggregates assemble into larger aggregates with more complex structures.

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publication status
published
subject
in
International Journal of Pharmaceutics
volume
637
article number
122829
publisher
Elsevier
external identifiers
  • pmid:36948472
  • scopus:85151369842
ISSN
0378-5173
DOI
10.1016/j.ijpharm.2023.122829
language
English
LU publication?
yes
id
2e30ecb9-bbf3-4b61-8f98-62ff97343800
date added to LUP
2023-05-22 11:43:58
date last changed
2024-09-07 11:11:14
@article{2e30ecb9-bbf3-4b61-8f98-62ff97343800,
  abstract     = {{<p>Three orthogonal techniques were used to provide new insights into thermally induced aggregation of the therapeutic protein Somatropin at pH 5.8 and 7.0. The techniques were Dynamic Light Scattering (DLS), Asymmetric Flow-Field Flow-Fractionation (AF4), and the TEM-based analysis system MiniTEM™. In addition, Differential Scanning Calorimetry (DSC) was used to study the thermal unfolding and stability. DSC and DLS were used to explain the initial aggregation process and aggregation rate at the two pH values. The results suggest that less electrostatic stabilization seems to be the main reason for the faster initial aggregation at pH 5.8, i.e., closer to the isoelectric point of Somatropin. AF4 and MiniTEM were used to investigate the aggregation pathway further. Combining the results allowed us to demonstrate Somatropin's thermal aggregation pathway at pH 7.0. The growth of the aggregates appears to follow two steps. Smaller elongated aggregates are formed in the first step, possibly initiated by partly unfolded species. In the second step, occurring during longer heating, the smaller aggregates assemble into larger aggregates with more complex structures.</p>}},
  author       = {{Västberg, Amanda and Bolinsson, Hans and Leeman, Mats and Nilsson, Lars and Nylander, Tommy and Sejwal, Kushal and Sintorn, Ida Maria and Lidayová, Kristina and Sjögren, Helen and Wahlgren, Marie and Elofsson, Ulla}},
  issn         = {{0378-5173}},
  language     = {{eng}},
  publisher    = {{Elsevier}},
  series       = {{International Journal of Pharmaceutics}},
  title        = {{Investigating thermally induced aggregation of Somatropin- new insights using orthogonal techniques}},
  url          = {{http://dx.doi.org/10.1016/j.ijpharm.2023.122829}},
  doi          = {{10.1016/j.ijpharm.2023.122829}},
  volume       = {{637}},
  year         = {{2023}},
}