Soliman Mahmoud G, Martinez-Serra Alberto, Dobricic Marko, Trinh Duong N, Cheeseman Jack, Spencer Daniel I R, Monopoli Marco P
Chemistry Department, RCSI (Royal College of Surgeons in Ireland), Dublin, Ireland.
Physics Department, Faculty of Science, Al-Azhar University, Nasr City, Egypt.
Front Toxicol. 2024 Jul 23;6:1393330. doi: 10.3389/ftox.2024.1393330. eCollection 2024.
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.
工程纳米颗粒（NPs）具有一系列有趣的特性，这使得它们在许多应用中都很有用。然而，持续接触纳米颗粒需要深入了解这些纳米颗粒与不同生物环境相互作用时的结果。在暴露于这些环境之后，纳米颗粒的原始表面会与周围介质中的分子强烈相互作用，这些分子包括代谢物、脂质、聚糖和蛋白质，从而形成所谓的蛋白质冠（PC）。众所周知，纳米颗粒-蛋白质冠会强烈影响各种纳米颗粒类型的生物学命运，包括细胞摄取、毒性和生物分布。因此，为了正确评估与工程纳米颗粒相关的潜在危害，研究和评估纳米颗粒周围形成的蛋白质冠是必不可少的。在此，我们详细描述了纳米颗粒-蛋白质冠复合物的分离和表征方案，并涵盖以下方面：1）在一系列暴露介质中针对不同纳米材料的分离方案，包括用于超顺磁性纳米颗粒的磁分离方法；2）使用常规实验室中可用的先进和标准技术对纳米颗粒进行物理化学表征；3）对蛋白质和聚糖成分进行纳米颗粒-蛋白质冠表征。
