Attempts to preserve and visualize protein corona on the surface of biological nanoparticles in blood serum using photomodification

利用光修饰技术在血清中保存和可视化生物纳米颗粒表面的蛋白质冠状层的尝试

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Abstract

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.

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