Fig. 1. General process for preparation of protein corona and common sources of errors and misinterpretation.

General steps for preparation of protein corona in vitro or ex vivo: preparation of nanoparticles and biological fluids; mixing of nanoparticle and biological fluids; incubating the mixture for a certain time and at a specific temperature; isolation of protein corona–coated nanoparticles using the five most common approaches (i.e., centrifugation-based, gradient centrifugation, size exclusion chromatography, magnetic separation, and field-flow fractionation); purification process; and protein corona characterization by proteomic techniques. Red text shows the common methodological features that may cause errors in the outcomes of proteomics analysis of the protein corona (e.g., considering protein impurities in the proteomics data).