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Spray drying for protein stabilization.

Katharina Tatjana Kopp1, Maarten De Beer2, Jody Voorspoels2

  • 1Eurofins Amatsigroup, Industriepark-Zwijnaarde 7B, 9052 Gent, Belgium; Drug Delivery and Disposition, KU Leuven, Department of Pharmaceutical and Pharmacological Sciences, Campus Gasthuisberg ON2, Herestraat 49, 3000 Leuven, Belgium.

International Journal of Pharmaceutics
|April 25, 2025
PubMed
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This study evaluated protein stabilization strategies for therapeutic proteins using spray drying (SD). Differential Scanning Fluorimetry (DSF) and Static Light Scattering (SLS) predicted stability, but results varied across proteins, questioning the sufficiency of these methods for solidification.

Area of Science:

  • Biopharmaceutical Formulation
  • Protein Stabilization Technologies
  • Analytical Chemistry

Background:

  • Therapeutic proteins require stabilization against inherent instability during formulation.
  • Solidification techniques like spray drying (SD) are employed to enhance protein stability.
  • Excipients and buffers are crucial components in developing stable protein formulations.

Purpose of the Study:

  • To assess the efficacy of selected buffers and excipients in enhancing the stability of three model proteins (α-chymotrypsin, catalase, Horseradish Peroxidase) during spray drying.
  • To compare the predictive capability of Differential Scanning Fluorimetry (DSF) combined with Static Light Scattering (SLS) for protein stability before and after spray drying.
  • To investigate the necessity of a holistic approach beyond DSF/SLS for predicting protein stability during solidification.
Keywords:
Differential scanning fluorimetryExcipientsProtein stabilitySpray dryingStatic light scattering

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Main Methods:

  • Formulation development using selected buffers and excipients for three protein concepts per model protein.
  • Differential Scanning Fluorimetry (DSF) and Static Light Scattering (SLS) for assessing protein stability in solution.
  • Spray drying (SD) of formulated proteins.
  • Post-SD stability assessment using dynamic light scattering, UV-VIS spectroscopy, far-UV circular dichroism, size-exclusion chromatography, and reversed-phase chromatography.

Main Results:

  • Significant variations in protein recovery and stability were observed after spray drying, with some proteins experiencing over 40% loss in concentration.
  • Only one formulation concept for catalase successfully maintained its original protein concentration post-SD.
  • DSF/SLS accurately predicted the stability of two catalase formulations but failed for the other model proteins, indicating limitations in predictive power.
  • Excipient selection showed minimal differences compared to previous studies on other proteins like BSA, IgG, and lysozyme.

Conclusions:

  • The effectiveness of selected excipients and buffers for protein stabilization during spray drying varied significantly among different proteins.
  • Differential Scanning Fluorimetry (DSF) and Static Light Scattering (SLS) show potential for predicting protein stability but may not be sufficient on their own for all proteins undergoing solidification.
  • A more comprehensive, holistic analytical strategy is likely required to accurately predict and ensure therapeutic protein stability after spray drying.