Journal Article

Analysis of nanomaterial biocoronas in biological and environmental surroundings

Zhang, Peng, Reilly, Katie, Ellis, Laura-Jayne A., Zhang, Wei, Martinez, Diego Stéfani Teodoro, Abdolahpur Monikh, Fazel, Wheeler, Korin E., Ramautar, Rawi, Chetwynd, Andrew J., Faserl, Klaus, Guo, Zhiling, Chen, Chunying, Lynch, Iseult, Cai, Rong, Cao, Mingjing, Davoudi, Hossein Hayat

2024

Overview

A biomolecular coating, or biocorona, forms on the surface of engineered nanomaterials (ENMs) immediately as they enter biological or environmental systems, defining their biological and environmental identity and influencing their fate and performance. This biomolecular layer includes proteins (the protein corona) and other biomolecules, such as nucleic acids and metabolites. To ensure a meaningful and reproducible analysis of the ENMs-associated biocorona, it is essential to streamline procedures for its preparation, separation, identification and characterization, so that studies in different labs can be easily compared, and the information collected can be used to predict the composition, dynamics and properties of biocoronas acquired by other ENMs. Most studies focus on the protein corona as proteins are easier to monitor and characterize than other biomolecules and play crucial roles in receptor engagement and signaling; however, metabolites play equally critical roles in signaling. Here we describe how to reproducibly prepare and characterize biomolecule-coated ENMs, noting especially the steps that need optimization for different types of ENMs. The structure and composition of the biocoronas are characterized using general methods (transmission electron microscopy, dynamic light scattering, capillary electrophoresis–mass spectrometry and liquid chromatography–mass spectrometry) as well as advanced techniques, such as transmission electron cryomicroscopy, synchrotron-based X-ray absorption near edge structure and circular dichroism. We also discuss how to use molecular dynamic simulation to study and predict the interaction between ENMs and biomolecules and the resulting biocorona composition. The application of this protocol can provide mechanistic insights into the formation, composition and evolution of the ENM biocorona, ultimately facilitating the biomedical and agricultural application of ENMs and a better understanding of their impact in the environment. Key points The authors provide a detailed workflow for the isolation and biophysical characterization of biomolecule corona (biocorona) components (proteins and metabolites) through mass spectrometry, advanced structural techniques (for example, transmission electron cryomicroscopy and synchrotron-based X-ray absorption near edge structure) and molecular dynamic simulations to model ENM–biocorona interactions. The designed pipeline normalizes the acquisition of data in different labs, increases their reproducibility, and facilitates their use for the prediction of the biocoronas acquired by less characterized ENMs. Engineered and anthropogenic nanoscale materials in the environment acquire a coating of biomolecules (biocorona) that modulates their properties, uptake and biodistribution. This protocol streamlines biocorona analysis to support the development of safe and sustainable nanotechnology.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 631/61/350/354 / 631/61/54/1754 / Absorption / Analytical Chemistry / Biological Techniques / Biomedical and Life Sciences / Biomolecules / Capillary electrophoresis / Chromatography, Liquid - methods / Circular dichroism / Computational Biology/Bioinformatics / Data acquisition / Dichroism / Electrons / Electrophoresis / Electrophoresis, Capillary - methods / Life Sciences / Light scattering / Liquid chromatography / Mass spectrometry / Mass Spectrometry - methods / Mass spectroscopy / Metabolites / Microarrays / Microscopy, Electron, Transmission - methods / Molecular dynamics / Molecular structure / Nanomaterials / Nanostructures - chemistry / Nanotechnology / Nucleic acids / Organic Chemistry / Photon correlation spectroscopy / Protein composition / Protein Corona - analysis / Protein Corona - chemistry / Protein Corona - metabolism / Proteins / Protocol / Reproducibility / Scientific imaging / Transmission electron microscopy / Workflow / X ray absorption