Automation and low-cost proteomics for characterization of the protein corona: experimental methods for big data
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RESEARCH PAPER
Automation and low-cost proteomics for characterization of the protein corona: experimental methods for big data Karsten M. Poulsen 1 & Thomas Pho 2,3 & Julie A. Champion 2,3 & Christine K. Payne 1 Received: 16 February 2020 / Revised: 13 May 2020 / Accepted: 19 May 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract 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. Keywords Biomaterials . Nanoparticles/nanotechnology . Spectroscopy/instrumentation
Introduction Nanoparticles (NPs) used in any “real-world” application encounter a range of molecules that will adsorb on the surface of the NP [1–10]. These adsorbed molecules, referred to as a
Published in the topical collection featuring Female Role Models in Analytical Chemistry. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00216-020-02726-1) contains supplementary material, which is available to authorized users. * Julie A. Champion [email protected] * Christine K. Payne [email protected] 1
Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA
2
School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA
3
Parker H. Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, GA 30332, USA
“corona,” determine how the NP functions in downstream applications. The majority of studies in this area have focused on the protein corona that forms when NPs, often developed for nanomedicine applications, interact with blood serum proteins. This protein corona is responsible for subsequent NPcell interactions including NP clearance from circulation and accumulati
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