Institute for Immunology, University Medical Center of the Johannes-Gutenberg University Mainz, Langenbeckstrasse 1, 55101 Mainz, Germany.
ACS Nano. 2011 Sep 27;5(9):7155-67. doi: 10.1021/nn201950e. Epub 2011 Aug 25.
In biological fluids, proteins associate with nanoparticles, leading to a protein "corona" defining the biological identity of the particle. However, a comprehensive knowledge of particle-guided protein fingerprints and their dependence on nanomaterial properties is incomplete. We studied the long-lived ("hard") blood plasma derived corona on monodispersed amorphous silica nanoparticles differing in size (20, 30, and 100 nm). Employing label-free liquid chromatography mass spectrometry, one- and two-dimensional gel electrophoresis, and immunoblotting the composition of the protein corona was analyzed not only qualitatively but also quantitatively. Detected proteins were bioinformatically classified according to their physicochemical and biological properties. Binding of the 125 identified proteins did not simply reflect their relative abundance in the plasma but revealed an enrichment of specific lipoproteins as well as proteins involved in coagulation and the complement pathway. In contrast, immunoglobulins and acute phase response proteins displayed a lower affinity for the particles. Protein decoration of the negatively charged particles did not correlate with protein size or charge, demonstrating that electrostatic effects alone are not the major driving force regulating the nanoparticle-protein interaction. Remarkably, even differences in particle size of only 10 nm significantly determined the nanoparticle corona, although no clear correlation with particle surface volume, protein size, or charge was evident. Particle size quantitatively influenced the particle's decoration with 37% of all identified proteins, including (patho)biologically relevant candidates. We demonstrate the complexity of the plasma corona and its still unresolved physicochemical regulation, which need to be considered in nanobioscience in the future.
在生物流体中，蛋白质与纳米颗粒结合，形成了定义颗粒生物学特性的蛋白质“冠”。然而，对于颗粒引导的蛋白质指纹及其对纳米材料特性的依赖，我们的了解还不全面。我们研究了单分散无定形二氧化硅纳米颗粒（粒径分别为 20、30 和 100nm）在长期存在的（“硬”）血液血浆衍生的冠层上的情况。采用无标记液相色谱-质谱联用、一维和二维凝胶电泳以及免疫印迹技术，不仅定性而且定量地分析了蛋白质冠层的组成。根据蛋白质的物理化学和生物学特性，对检测到的蛋白质进行了生物信息学分类。被鉴定的 125 种蛋白质的结合并不简单地反映了它们在血浆中的相对丰度，而是揭示了特定脂蛋白以及参与凝血和补体途径的蛋白质的富集。相比之下，免疫球蛋白和急性期反应蛋白对颗粒的亲和力较低。带负电荷的颗粒的蛋白质修饰与蛋白质的大小或电荷无关，这表明静电效应本身并不是调节纳米颗粒-蛋白质相互作用的主要驱动力。值得注意的是，即使粒径差异仅为 10nm，也会显著决定纳米颗粒的冠层，尽管与颗粒表面体积、蛋白质大小或电荷没有明显的相关性。粒径定量地影响了颗粒的修饰，其中包括 37%的所有鉴定出的蛋白质，包括（病理）生物学相关的候选物。我们证明了血浆冠层的复杂性及其仍未解决的物理化学调节，这在未来的纳米生物科学中需要加以考虑。
