Digiacomo Luca, Giulimondi Francesca, Capriotti Anna Laura, Piovesana Susy, Montone Carmela Maria, Chiozzi Riccardo Zenezini, Laganà Aldo, Mahmoudi Morteza, Pozzi Daniela, Caracciolo Giulio
Department of Molecular Medicine, Sapienza University of Rome Viale Regina Elena 291 00161 Rome Italy
Department of Chemistry, Sapienza University of Rome P.le A. Moro 5 00185 Rome Italy.
Nanoscale Adv. 2021 May 17;3(13):3824-3834. doi: 10.1039/d1na00211b. eCollection 2021 Jun 30.
In the past few years, characterization of the protein corona (PC) that forms around liposomal systems has gained increasing interest for the development of novel therapeutic and diagnostic technologies. At the crossroads of fast-moving research fields, the interdisciplinarity of protein corona investigations poses challenges for experimental design and reporting. Isolation of liposome-protein complexes from biological fluids has been identified as a fundamental step of the entire workflow of PC characterization but exact specifications for conditions to optimize pelleting remain elusive. In the present work, key factors affecting precipitation of liposome-protein complexes by centrifugation, including time of centrifugation, total sample volume, lipid : protein ratio and contamination from biological NPs were comprehensively evaluated. Here we show that the total amount of isolated liposome-protein complexes and the extent of contamination from biological NPs may vary with influence factors. Our results provide protein corona researchers with precise indications to separate liposome-protein complexes from protein-rich fluids and include proper controls, thus they are anticipated to catalyze improved consistency of data mining and computational modelling of protein corona composition.
在过去几年中，脂质体系统周围形成的蛋白质冠层（PC）的表征对于新型治疗和诊断技术的开发越来越受到关注。在快速发展的研究领域的交叉点上，蛋白质冠层研究的跨学科性给实验设计和报告带来了挑战。从生物流体中分离脂质体 - 蛋白质复合物已被确定为蛋白质冠层表征整个工作流程的基本步骤，但优化沉淀条件的确切规范仍然难以捉摸。在本工作中，全面评估了影响脂质体 - 蛋白质复合物通过离心沉淀的关键因素，包括离心时间、总样品体积、脂质与蛋白质比例以及来自生物纳米颗粒的污染。我们在此表明，分离出的脂质体 - 蛋白质复合物的总量以及来自生物纳米颗粒的污染程度可能会因影响因素而有所不同。我们的结果为蛋白质冠层研究人员提供了从富含蛋白质的流体中分离脂质体 - 蛋白质复合物的精确指示，并包括适当的对照，因此预计它们将促进蛋白质冠层组成的数据挖掘和计算建模的一致性提高。
