Automation and low-cost proteomics for characterization of the protein corona: experimental methods for big data.

Published

Journal Article

Nanoparticles used in biological settings are exposed to proteins that adsorb on the surface forming a protein corona. These adsorbed proteins dictate the subsequent cellular response. A major challenge has been predicting what proteins will adsorb on a given nanoparticle surface. Instead, each new nanoparticle and nanoparticle modification must be tested experimentally to determine what proteins adsorb on the surface. We propose that any future predictive ability will depend on large datasets of protein-nanoparticle interactions. As a first step towards this goal, we have developed an automated workflow using a liquid handling robot to form and isolate protein coronas. As this workflow depends on magnetic separation steps, we test the ability to embed magnetic nanoparticles within a protein nanoparticle. These experiments demonstrate that magnetic separation could be used for any type of nanoparticle in which a magnetic core can be embedded. Higher-throughput corona characterization will also require lower-cost approaches to proteomics. We report a comparison of fast, low-cost, and standard, slower, higher-cost liquid chromatography coupled with mass spectrometry to identify the protein corona. These methods will provide a step forward in the acquisition of the large datasets necessary to predict nanoparticle-protein interactions.

Full Text

Duke Authors

Cited Authors

  • Poulsen, KM; Pho, T; Champion, JA; Payne, CK

Published Date

  • June 4, 2020

Published In

PubMed ID

  • 32500258

Pubmed Central ID

  • 32500258

Electronic International Standard Serial Number (EISSN)

  • 1618-2650

International Standard Serial Number (ISSN)

  • 1618-2642

Digital Object Identifier (DOI)

  • 10.1007/s00216-020-02726-1

Language

  • eng