Accessing to the minor proteome of red blood cells through the influence of the nanoparticle surface properties on the corona composition

Abstract

Affif Zaccaria,1,* Florence Roux-Dalvai,2,3,* Ali Bouamrani,1 Adrien Mombrun,1 Pascal Mossuz,4 Bernard Monsarrat,2,3 François Berger1 1Clinatec CEA-LETI, Grenoble, 2CNRS, IPBS (Institut de Pharmacologie et de Biologie Structurale), 3Université de Toulouse, UPS, IPBS, Toulouse, 4TIMC-Therex UMR 5525 CNRS, UJF, CHU Grenoble, Grenoble, France *These authors contributed equally to this work Abstract: Nanoparticle (NP)–protein interactions in complex samples have not yet been clearly understood. Nevertheless, several studies demonstrated that NP’s physicochemical features significantly impact on the protein corona composition. Taking advantage of the NP potential to harvest different subsets of proteins, we assessed for the first time the capacity of three kinds of superparamagnetic NPs to highlight the erythrocyte minor proteome. Using both qualitative and quantitative proteomics approaches, nano-liquid chromatography–tandem mass spectrometry allowed the identification of 893 different proteins, confirming the reproducible capacity of NPs to increase the number of identified proteins, through a reduction of the sample concentration range and the capture of specific proteins on the three different surfaces. These NP-specific protein signatures revealed significant differences in their isoelectric point and molecular weight. Moreover, this NP strategy offered a deeper access to the erythrocyte proteome highlighting several signaling pathways implicated in important erythrocyte functions. The automated potentiality, the reproducibility, and the low-consuming sample demonstrate the strong compatibility of our strategy for large-scale clinical studies and may become a standardized sample preparation in future erythrocyte-associated proteomics studies. Keywords: nanoparticles, red blood cells, mass spectrometry, quantitative proteomics, protein corona, minor proteome