Capillary electrophoresis mass spectrometry approaches for characterization of the protein and metabolite corona acquired by nanomaterials

Andrew J. Chetwynd*, Wei Zhang, Klaus Faserl, James A. Thorn, Iseult Lynch, Rawi Ramautar, Herbert H. Lindner

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


The adsorption of biomolecules from surrounding biological matrices to the surface of nanomaterials (NMs) to form the corona has been of interest for the past decade. Interest in the bio-nano interface arises from the fact that the biomolecular corona confers a biological identity to NMs and thus causes the body to identify them as “self”. For example, previous studies have demonstrated that the proteins in the corona are capable of interacting with membrane receptors to influence cellular uptake and established that the corona is responsible for cellular trafficking of NMs and their eventual toxicity. To date, most research has focused upon the protein corona and overlooked the possible impacts of the metabolites included in the corona or synergistic effects between components in the complete biomolecular corona. As such, this work demonstrates methodologies to characterize both the protein and metabolite components of the biomolecular corona using bottom-up proteomics and metabolomics approaches in parallel. This includes an on-particle digest of the protein corona with a surfactant used to increase protein recovery, and a passive characterization of the metabolite corona by analyzing metabolite matrices before and after NM exposures. This work introduces capillary electrophoresis – mass spectrometry (CESI-MS) as a new technique for NM corona characterization. The protocols outlined here demonstrate how CESI-MS can be used for the reliable characterization of both the protein and metabolite corona acquired by NMs. The move to CESI-MS greatly decreases the volume of sample required (compared to traditional liquid chromatography – mass spectrometry (LC-MS) approaches) with multiple injections possible from as little as 5 µL of sample, making it ideal for volume limited samples. Furthermore, the environmental consequences of analysis are reduced with respect to LC-MS due to the low flow rates (<20 nL/min) in CESI-MS, and the use of aqueous electrolytes which eliminates the need for organic solvents.

Original languageEnglish
Article numbere61760
JournalJournal of Visualized Experiments
Issue number164
Publication statusPublished - Oct 2020

Bibliographical note

Funding Information:
A.J.C, J.A.T and I.L acknowledge funding from the European Commission via Horizon 2020 project ACEnano (Grant no. H2020-NMBP-2016-720952). W.Z acknowledges PhD funding from the China Scholarship Council (CSC, No. 201507060011). R.R acknowledges the financial support of the Vidi grant scheme of the Netherlands Organization of Scientific Research (NWO Vidi 723.0160330).

Publisher Copyright:
© 2020, Journal of Visualized Experiments. All rights reserved.

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)


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