Physical-Chemical Aspects of Protein Corona: Relevance to in Vitro and in Vivo Biological Impacts of Nanoparticles
Physical-Chemical Aspects of Protein Corona: Relevance to in Vitro and in Vivo Biological Impacts of Nanoparticles
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DOI：
10.1021/ja107583h
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发表时间：
2011-03-02
影响因子：
15
通讯作者：
Dawson, Kenneth A.
中科院分区：
文献类型：
--
作者：
Monopoli, Marco P.;Walczyk, Dorota;Dawson, Kenneth A.
现在很明显，除了尺寸和形状，生物介质中纳米级物体的另一个主要定义元素是它们的长寿命蛋白质（“硬”）冠。这种冠可以表示为纳米颗粒（NP）单体的裸露表面的持久的稳定涂层，或者它可以反映在颗粒组装体的不同亚群中，每个亚群呈现持久的蛋白质涂层。使用的方法和物理化学的概念，我们涉及的研究在不同的血浆浓度的蛋白质冠的组合物的结构数据的复合物在原位和免费从多余的血浆。这使得在生物学背景下对硬蛋白质冠的含义具有高度的置信度。在这里，我们提出了两个组成不同的纳米粒子，即磺化聚苯乙烯和二氧化硅纳米粒子的蛋白质吸附。NP-蛋白质复合物的特征在于差示离心沉降、动态光散射和zeta电位，其在原位和一旦从血浆中分离作为蛋白质/NP表面积比的函数。然后，我们介绍了一个半定量测定他们的硬电晕组合物，使用一维十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和电喷雾液相色谱质谱，这使我们能够遵循的颗粒的总结合等温线，同时确定的性质和量的最相关的蛋白质作为血浆浓度的函数。我们发现，硬冠可以演变相当显着的蛋白质浓度，适合在体外细胞研究，目前在体内的研究，这有很深的影响，在体外体内外推，将需要在未来的一些考虑。
It is now clearly emerging that besides size and shape, the other primary defining element of nanoscale objects in biological media is their long-lived protein ("hard") corona. This corona may be expressed as a durable, stabilizing coating of the bare surface of nanoparticle (NP) monomers, or it may be reflected in different subpopulations of particle assemblies, each presenting a durable protein coating. Using the approach and concepts of physical chemistry, we relate studies on the composition of the protein corona at different plasma concentrations with structural data on the complexes both in situ and free from excess plasma. This enables a high degree of confidence in the meaning of the hard protein corona in a biological context. Here, we present the protein adsorption for two compositionally different NPs, namely sulfonated polystyrene and silica NPs. NP-protein complexes are characterized by differential centrifugal sedimentation, dynamic light scattering, and zeta-potential both in situ and once isolated from plasma as a function of the protein/NP surface area ratio. We then introduce a semiquantitative determination of their hard corona composition using one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrospray liquid chromatography mass spectrometry, which allows us to follow the total binding isotherms for the particles, identifying simultaneously the nature and amount of the most relevant proteins as a function of the plasma concentration. We find that the hard corona can evolve quite significantly as one passes from protein concentrations appropriate to in vitro cell studies to those present in in vivo studies, which has deep implications for in vitro in vivo extrapolations and will require some consideration in the future.