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PreprintCC BYChemRxiv

In-Situ and Label-Free Quantification of BSA Adsorption on Mesoporous Silica Nanoparticles via Orthogonal Analysis in Analytical Ultracentrifugation

  • Chao Zhang1,2
  • Mengdi Chen1,2
  • Bingwen Hu3,2
  • E Wu1,2,3,4,5,6,7,8
8 affiliations
  1. 1State Key Laboratory of Precision Spectroscopy
  2. 2East China Normal University
  3. 3Department of Physics and Shanghai Key Laboratory for Magnetic Resonance
  4. 4Institute of Magnetic Resonance and Molecular Imaging in Medicine
  5. 5Chongqing Key Laboratory of Precision Optics
  6. 6Chongqing Institute of East China Normal University
  7. 7Collaborative Innovation Center of Extreme Optics
  8. 8Shanxi University
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Abstract

Understanding the bio–nano interface requires precise, in-situ and label-free tools to characterize the protein corona. While hydrodynamic techniques like dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA) are widely used, they face inherent limitations: DLS provides ensemble-averaged signals and often requires specific optical models, whereas NTA’s temporal resolution limits its accuracy for ultra-small particles. Here, we demonstrate the power of custom-built multiwavelength analytical ultracentrifugation (MWL-AUC) as a superior orthogonal platform to characterize the bovine serum albumin (BSA) corona on mesoporous silica nanoparticles (MSNs). By integrating three independent analytical dimensions, i.e. in-situ supernatant depletion, density-based sedimentation analysis, and multiwavelength spectral decoupling, we achieved precise quantification of both the total adsorbed BSA and its spatial distribution. This approach enables a quantitative mapping of protein localization that transcends mere qualitative observations of aggregation. Specifically, we show that the compositional information from spectral decoupling provides a robust cross-validation for the effective density analysis, which fundamentally dictates the precision of the derived spatial distribution. This study establishes a self-validating framework for the precise characterization of protein corona on porous nanostructures, offering vital insights for the rational design of nanocarriers in complex biological environments.

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Cite

BibTeXRIS
BibTeX
@misc{zhang2026insitu,
  title = {{In-Situ and Label-Free Quantification of BSA Adsorption on Mesoporous Silica Nanoparticles via Orthogonal Analysis in Analytical Ultracentrifugation}},
  author = {Chao Zhang and Mengdi Chen and Bingwen Hu and E Wu},
  howpublished = {ChemRxiv},
  year = {2026},
  month = sep,
  doi = {10.26434/chemrxiv.15009116/v1},
  url = {https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009116/v1},
  note = {CC BY},
}
RIS
TY  - UNPB
TI  - In-Situ and Label-Free Quantification of BSA Adsorption on Mesoporous Silica Nanoparticles via Orthogonal Analysis in Analytical Ultracentrifugation
AU  - Chao Zhang
AU  - Mengdi Chen
AU  - Bingwen Hu
AU  - E Wu
PY  - 2026
DA  - 2026/09/20
PB  - ChemRxiv
DO  - 10.26434/chemrxiv.15009116/v1
UR  - https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009116/v1
AB  - Understanding the bio–nano interface requires precise, in-situ and label-free tools to characterize the protein corona. While hydrodynamic techniques like dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA) are widely used, they face inherent limitations: DLS provides ensemble-averaged signals and often requires specific optical models, whereas NTA’s temporal resolution limits its accuracy for ultra-small particles. Here, we demonstrate the power of custom-built multiwavelength analytical ultracentrifugation (MWL-AUC) as a superior orthogonal platform to characterize the bovine serum albumin (BSA) corona on mesoporous silica nanoparticles (MSNs). By integrating three independent analytical dimensions, i.e. in-situ supernatant depletion, density-based sedimentation analysis, and multiwavelength spectral decoupling, we achieved precise quantification of both the total adsorbed BSA and its spatial distribution. This approach enables a quantitative mapping of protein localization that transcends mere qualitative observations of aggregation. Specifically, we show that the compositional information from spectral decoupling provides a robust cross-validation for the effective density analysis, which fundamentally dictates the precision of the derived spatial distribution. This study establishes a self-validating framework for the precise characterization of protein corona on porous nanostructures, offering vital insights for the rational design of nanocarriers in complex biological environments.
ER  -