Mass spectrometric approach for the analysis of the hard protein corona of nanoparticles in living cells
| Autor: | Maria Montes-Bayón, Nerea Fernández-Iglesias, Gergo Peter Szekeres, Janina Kneipp, Jörg Bettmer |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
endocrine system Biophysics Metal Nanoparticles Nanoparticle Breast Neoplasms Protein Corona Living cell Biochemistry Mass Spectrometry Cell Line Mice 03 medical and health sciences Corona (optical phenomenon) Animals Humans Gel electrophoresis 030102 biochemistry & molecular biology Chemistry Macrophages Mass spectrometric Electrophoresis 030104 developmental biology Colloidal gold Female Gold |
| Zdroj: | Journal of Proteomics. 212:103582 |
| ISSN: | 1874-3919 |
| Popis: | The diagnostic and therapeutic application of nanoparticles requires comprehensive knowledge of their interaction with the biomolecular surroundings. The formation of the protein corona on nanoparticles that were internalized by living cells is yet to be understood. In this study, we present a robust approach for the electrophoretic and mass spectrometric analysis of the hard protein corona composition formed in living cells on ~30 nm citrate-stabilized gold nanoparticles, i.e., the proteins with the highest affinity towards the gold nanoparticle surface. The gold nanoparticles were internalized by MCF-7 cells for 24 h followed by the extraction of the hard protein corona‑gold nanoparticle bioconjugates from living cell cultures. The extracted proteins were then separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and analyzed by ESI-Q-TOF-MS, which allowed to identify 108 hard corona proteins. The experiments were repeated with J774 macrophage cells with incubation times of 1.5 h, 3 h, 6 h, and 24 h, and the results showed that the hard protein corona remained unchanged over time. Therefore, the proposed experimental approach proved to be a valuable tool for identifying hard corona proteins of nanoparticles internalized by living cells. |
| Databáze: | OpenAIRE |
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