AUTHOR=Soliman Mahmoud G. , Martinez-Serra Alberto , Dobricic Marko , Trinh Duong N. , Cheeseman Jack , Spencer Daniel I. R. , Monopoli Marco P. TITLE=Protocols for isolation and characterization of nanoparticle biomolecular corona complexes JOURNAL=Frontiers in Toxicology VOLUME=6 YEAR=2024 URL=https://www.frontiersin.org/journals/toxicology/articles/10.3389/ftox.2024.1393330 DOI=10.3389/ftox.2024.1393330 ISSN=2673-3080 ABSTRACT=

Engineered nanoparticles (NPs) pose a broad spectrum of interesting properties that make them useful for many applications. However, continuous exposure to NPs requires the need to deeply understand the outcomes when these NPs interact with different biological environments. After exposure within (to) these environments, the pristine surfaces of NPs strongly interact with the molecules from the surrounding medium, including metabolites, lipids, glycan, and proteins, forming the so-called protein corona (PC). It is well established that the NP-PC strongly influences the biological fate of various NPs types, including cellular uptake, toxicity, and biodistribution. Thus, for a proper assessment of potential hazards associated with engineered NPs, it is mandatory to study and evaluate the PC that forms around NPs. Herein, we describe protocols in detail for the isolation and characterization of NP-PC complexes and cover the following aspects: 1) isolation protocols for different nanomaterials in a range of exposing media, including magnetic isolation methods for superparamagnetic NPs, 2) NP physico-chemical characterization using advanced and standard techniques available in regular laboratories, and 3) NP- PC characterization of the protein and glycan components.