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Proteomic and Lipidomic Analysis of Nanoparticle Corona upon Contact with Lung Surfactant Reveals Differences in Protein, but Not Lipid Composition

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Department of Pharmacy, Saarland University, 66123 Saarbruecken, Germany
HIPS − Helmholtz Institute for Pharmaceutical Research Saarland, Helmholtz Centre for Infection Research, 66123 Saarbruecken, Germany
§ Institute of Immunology, Mainz University, 55131 Mainz, Germany
Scientific Consilience GmbH, Saarland University, 66123 Saarbruecken, Germany
Department of Biochemistry and Molecular Biology, Faculty of Biology, Complutense University, 28040 Madrid, Spain
*Address correspondence to [email protected]
Cite this: ACS Nano 2015, 9, 12, 11872–11885
Publication Date (Web):November 16, 2015
Copyright © 2015 American Chemical Society

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    Pulmonary surfactant (PS) constitutes the first line of host defense in the deep lung. Because of its high content of phospholipids and surfactant specific proteins, the interaction of inhaled nanoparticles (NPs) with the pulmonary surfactant layer is likely to form a corona that is different to the one formed in plasma. Here we present a detailed lipidomic and proteomic analysis of NP corona formation using native porcine surfactant as a model. We analyzed the adsorbed biomolecules in the corona of three NP with different surface properties (PEG-, PLGA-, and Lipid-NP) after incubation with native porcine surfactant. Using label-free shotgun analysis for protein and LC–MS for lipid analysis, we quantitatively determined the corona composition. Our results show a conserved lipid composition in the coronas of all investigated NPs regardless of their surface properties, with only hydrophilic PEG-NPs adsorbing fewer lipids in total. In contrast, the analyzed NP displayed a marked difference in the protein corona, consisting of up to 417 different proteins. Among the proteins showing significant differences between the NP coronas, there was a striking prevalence of molecules with a notoriously high lipid and surface binding, such as, e.g., SP-A, SP-D, DMBT1. Our data indicate that the selective adsorption of proteins mediates the relatively similar lipid pattern in the coronas of different NPs. On the basis of our lipidomic and proteomic analysis, we provide a detailed set of quantitative data on the composition of the surfactant corona formed upon NP inhalation, which is unique and markedly different to the plasma corona.

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    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acsnano.5b04215.

    • Scheme of pPS preparation; Dynamic light scattering of pPS membranes; Comparison of separation methods; Most abundant lipid species in pPS and the corona of NPs; Adsorption of Alveofact to NPs; Comparison of protein adsorption by means of MW, IEP, and Gravy Score; Characterization of NPs. (PDF)

    • List of all proteins and lipids that were determined in pPS and in the corona of NPs. (XLSX)

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