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. 2025 Jul 15;97(27):14132-14141.
doi: 10.1021/acs.analchem.4c05573. Epub 2025 Jun 30.

Multiple Exposures of Plasma to Nanoparticles: A Novel Tool to Personalize Biomolecular Coronas and Fractionate Fluids

Alberto Martinez-Serra  1 , Jack Cheeseman  2 , Asia Saorin  1 , Mahmoud G Soliman  1 , Marko Dobricic  1 , Daniel I R Spencer  2 , Marco P Monopoli  1
Affiliations

Multiple Exposures of Plasma to Nanoparticles: A Novel Tool to Personalize Biomolecular Coronas and Fractionate Fluids

Alberto Martinez-Serra et al. Anal Chem. .

Abstract

Nanoparticles (NPs) have emerged as a valuable tool for biomarker discovery due to their ability to interact with biological fluids and form biomolecular coronas. In this study, we introduce a multiple exposure method that uses NPs to fractionate biological fluids and obtain personalized coronas. By repeatedly exposing plasma to silica NPs, we observed a progressive change of the biomolecule profile in both the pellet and supernatant. The varying protein and glycan composition of the corona was characterized using techniques such as SDS-PAGE, mass spectrometry, and UHPLC. Notably, the corona's composition evolved with each exposure cycle, reflecting the selective binding of proteins and glycosylated molecules from a corona of high-affinity biomolecules to a more diverse corona with very distinct structures. By tracing the sequential modification of protein and glycan composition, we believe that the method can be useful to trace specific biomarker profiles, offering a noninvasive alternative to conventional diagnostic processes with the potential to become a useful tool for disease monitoring and advanced biomedical applications.

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Figures

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1
Schematic representation of the multiple exposure method to obtain tuned biocoronas and fractionated fluids. Plasma is incubated with NPs to form biomolecular coronas. Centrifugation separates NPs with a corona from the supernatant. Repeated incubation with fresh NPs produces coronas with distinct composition, yielding a “fractionated” plasma and “tuned” coronas. Created with Biorender.com.
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2
Physicochemical characterization of the pristine NPs and the NP-corona complexes. (a) TEM image of the silica NPs. Scale bar corresponds to 200 nm. (b) Hydrodynamic diameter from DLS of pristine and NPs with corona. (c) Apparent diameter from DCS of pristine and corona NPs. The formation of a corona is reflected in a shift to the left due to the change of density from the layer of biomolecules formed on the NP surface (inset).
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3
Comparative analysis of the protein profiles from the NP corona samples. (a) SDS–PAGE and (b) corresponding densitometry analysis for NP0–NP8. (c) Example of proteins identified in the corona by mass spectrometry (Table S1) changing their abundance in the NP corona through the different samples of the multiple exposure method. (t test, *p < 0.1).
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4
Comparative analysis of the glycan profiles from the NP corona samples. (a) UHPLC profiles of the glycan moieties embedded in the corona. (b) Example of glycan compositions changing their abundance in the NP corona over the different samples of the multiple exposure method. The glycan cartoons shown are for illustrative purposes only; precise features like isomer type would have to be confirmed by other analytical means (t test, *p < 0.05).
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5
Comparative analysis of protein and glycan profiles from plasma samples. (a) SDS–PAGE and (b) densitometry for the plasma and S8 after multiple exposure to NPs. (c) Chromatograms corresponding to the glycan profiles of plasma and S8. The glycan cartoons shown are for illustrative purposes only; precise features like isomer type would have to be confirmed by other analytical means.
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