Department of Chemistry, Fudan University, Shanghai 200433, China.
Zhejiang BioHarmonious SciTech. Co. Ltd, Hangzhou 310018, China.
Nanoscale. 2017 Apr 6;9(14):4787-4792. doi: 10.1039/c7nr01523b.
Nanoparticle (NP) surfaces are modified immediately by the adsorption of proteins when exposed to human blood, leading to the formation of a protein corona. The adsorption of serum proteins is the key process for exploring the bioapplication and biosafety of NPs. In this study, NP-protein binding affinity (K) was investigated. Some serum proteins, such as human serum albumin (HSA), trypsin (TRP), hemoglobin (Hb), myoglobin (MB), immunoglobulin G (IgG), carbonic anhydrase (CA), fibrinogen (FIB), chymotrypsin and r-globulin, were used with gold nanoparticles (AuNPs) to address binding affinity according to isothermal titration calorimetry (ITC) combined with dynamic light scattering (DLS) and fluorescence quenching. The NP protein binding affinities determined by the two methods were in agreement, and depended on the protein properties and size of the NPs. The two methods are convenient, and the results are highly comparable. These methods can be extended to determine the binding affinity of NP protein interactions. The adsorption of proteins upon the AuNP surface is a complex process and depends on several factors, but the binding affinities are higher for proteins with more cysteine residues located on the surface.
纳米粒子 (NP) 表面在暴露于人体血液时会立即被蛋白质吸附而发生修饰，从而形成蛋白质冠。血清蛋白的吸附是探索 NPs 的生物应用和生物安全性的关键过程。在本研究中，研究了 NP-蛋白质结合亲和力 (K)。使用金纳米粒子 (AuNP) 与一些血清蛋白（如人血清白蛋白 (HSA)、胰蛋白酶 (TRP)、血红蛋白 (Hb)、肌红蛋白 (MB)、免疫球蛋白 G (IgG)、碳酸酐酶 (CA)、纤维蛋白原 (FIB)、糜蛋白酶和 r-球蛋白）根据等温热力学滴定 (ITC) 结合动态光散射 (DLS) 和荧光猝灭法来研究结合亲和力。两种方法确定的 NP 蛋白质结合亲和力一致，并且取决于蛋白质特性和 NPs 的大小。这两种方法都很方便，结果高度可比。这些方法可以扩展到确定 NP 蛋白质相互作用的结合亲和力。蛋白质在 AuNP 表面的吸附是一个复杂的过程，取决于几个因素，但表面上带有更多半胱氨酸残基的蛋白质具有更高的结合亲和力。
