Poletaeva Julia E, Tupitsyna Anastasiya V, Grigor'eva Alina E, Dovydenko Ilya S, Ryabchikova Elena I
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Science, Lavrent'ev av., 8, Novosibirsk, 630090, Russian Federation.
Beilstein J Nanotechnol. 2024 Dec 30;15:1654-1666. doi: 10.3762/bjnano.15.130. eCollection 2024.
A protein corona is present on any nanoparticle (NP) entering biological fluids; however, the existence of a natural protein corona on natural NPs has not been experimentally confirmed. We used our previously developed photomodification method to fix the natural corona on "biological nanoparticles" (bio-NPs) in fetal bovine serum and newborn bovine serum; native sera served as a control. To isolate photomodified bio-NPs, we used ultracentrifugation (UC), sucrose gradient (12%, 30%, and 50%), and sucrose cushion (30%) methods. Isolated bio-NPs were visualized using transmission electron microscopy. The results showed that a protein corona is present on extracellular vesicles and lipoproteins isolated by UC. The isolation of bio-NPs through sucrose gradient or cushion did not preserve the protein corona. At the same time, we observed signs of a negative effect of the sucrose gradient on bio-NPs of intact and photomodified serum; when isolating these particles on a sucrose cushion, no negative effects were observed. We believe that the data we present will be useful to researchers using sucrose solutions to isolate bio-NPs and working on the properties of the protein corona. In this work, we have obtained direct images of a "natural" protein corona on natural bio-NPs of blood serum for the first time.
任何进入生物流体的纳米颗粒（NP）上都会形成蛋白质冠层；然而，天然纳米颗粒上天然蛋白质冠层的存在尚未得到实验证实。我们使用之前开发的光修饰方法将天然冠层固定在胎牛血清和新生牛血清中的“生物纳米颗粒”（生物NP）上；以天然血清作为对照。为了分离光修饰的生物NP，我们使用了超速离心（UC）、蔗糖梯度（12%、30%和50%）和蔗糖垫层（30%）方法。使用透射电子显微镜观察分离出的生物NP。结果表明，通过UC分离的细胞外囊泡和脂蛋白上存在蛋白质冠层。通过蔗糖梯度或垫层分离生物NP并不能保留蛋白质冠层。同时，我们观察到蔗糖梯度对完整血清和光修饰血清的生物NP有负面影响的迹象；当在蔗糖垫层上分离这些颗粒时，未观察到负面影响。我们相信，我们提供的数据将对使用蔗糖溶液分离生物NP并研究蛋白质冠层特性的研究人员有用。在这项工作中，我们首次获得了血清天然生物NP上“天然”蛋白质冠层的直接图像。
