基于二氧化硅胶体晶体薄膜和反射干涉光谱的蛋白冠监测方法

doi:10.6023/A20090422

摘要/Abstract

生物分子与纳米材料作用形成的“蛋白冠”影响纳米材料的物理和化学性质, 目前缺少有效的原位实时技术监测蛋白冠的形成过程. 本研究基于二氧化硅胶体晶体薄膜和反射干涉光谱法, 研究了三种代表性血液蛋白质在二氧化硅纳米粒子表面的蛋白冠形成过程, 结果表明这三种蛋白具有不同的蛋白冠形成过程及参数; 研究了人血清白蛋白在三种表面曲率的二氧化硅表面的蛋白冠形成过程, 结果表明曲率越大时, 蛋白冠形成速率越快, 厚度越大. 以血浆和全血样品为生物环境开展蛋白冠形成过程研究, 结果表明本研究的监测方法可以直接用于血浆和全血在纳米粒子表面蛋白冠的形成过程监测, 为纳米材料与生物分子的相互作用研究提供了一种简单可靠的评价技术.

关键词: 血浆蛋白, 蛋白冠, 反射干涉光谱法, 二氧化硅胶体晶体薄膜, 有序多孔层干涉测量技术

It will form protein corona on the surface of nanoparticles when exposed to the biological environment. The process of protein corona formation is crucial in biomedical engineering, nanomaterials development and research, especially the protein corona formation process of medical engineering materials in blood samples, and there is no mature method for real- time monitoring. Silica colloidal crystal films were prepared by vertical deposition on the glass slides. The glass slides were placed vertically in an ethanol suspension of silica nonospheres with diameters of 110, 190 and 280 nm. After 7 days of ethanol evaporation, a large area of silica colloidal crystal film was formed on the surface of the glass slide. The glass slides of silica colloidal crystal films were assembled into reaction cell, and the reflectometric interference spectroscopy (RIfS) system assembled by microscope light source and spectrometer were used for detection and optical thickness as a parameter. And different concentrations of hydrochloric acid were used to test the performance of the RIfS system. In this study, RIfS was utilized to detect the protein corona formation of human serum albumin, human fibrinogen and immunoglobulin G, as well as blood samples by silica colloidal crystal films which have special optical properties. It was found that the formation of immunoglobulin G had the highest optical thickness variation due to its conformation. According to the formation process of human serum albumin on silica nanosheres with different diameters, the optical thickness of protein corona was also higher on the of silica nanospheres with higher curvature. In addition, through the detection of plasma and whole blood samples, it was found that the silica colloidal crystal films could basically eliminate the interference of blood cells to the detection of protein corona. This work developed a real-time unlabeled protein corona research method, which provides a basis for the clinical application of biomedical materials.

Key words: plasma protein, protein corona, reflectometric interference spectroscopy, silica colloidal crystal film, ordered porous layer interferometry

引用此文

吴峰, 苏倩倩, 周乐乐, 许鹏飞, 董傲, 钱卫平. 基于二氧化硅胶体晶体薄膜和反射干涉光谱的蛋白冠监测方法[J]. 化学学报, 2021, 79(3): 338-343.

Feng Wu, Qianqian Su, Lele Zhou, Pengfei Xu, Ao Dong, Weiping Qian. A Novel Protein Corona Characterization based on the Reflectometric Interference Spectroscopy with Silica Colloidal Crystal Films[J]. Acta Chimica Sinica, 2021, 79(3): 338-343.

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图/表 8

图1. (a) 110 nm直径二氧化硅胶体晶体薄膜截面SEM图; (b) 110 nm直径二氧化硅胶体晶体薄膜俯视SEM图; (c) 190 nm直径二氧化硅胶体晶体薄膜截面SEM图; (d) 190 nm直径二氧化硅胶体晶体薄膜俯视SEM图; (e) 280 nm直径二氧化硅胶体晶体薄膜截面SEM图; (f) 280 nm直径二氧化硅胶体晶体薄膜俯视SEM图

Figure 1. (a) Sectional view SEM image of silica colloidal crystal film with a diameter of 110 nm silica spheres; (b) top view SEM image of silica colloidal crystal film with a diameter of 110 nm ; (c) sectional view SEM image of silica colloidal crystal film with a diameter of 190 nm; (d) top view SEM image of silica colloidal crystal film with a diameter of 190 nm; (e) sectional view SEM image of silica colloidal crystal film with a diameter of 280 nm; (f) top view SEM image of silica colloidal crystal film with a diameter of 280 nm

图2. (a)不同球体尺寸的二氧化硅胶体晶体薄膜的典型反射光谱; (b)不同球体尺寸的二氧化硅胶体晶体薄膜的照片, 由左到右分别为110, 190和280 nm

Figure 2. (a) Typical reflectance interference spectra silica colloidal crystal film of different sphere sizes; (b) photograph of silica colloidal crystal film of different sphere sizes, 110, 190 and 280 nm respectively from left to right

图3. 基于二氧化硅胶体晶体薄膜和反射干涉法的蛋白冠检测装置示意图

Figure 3. Schematic diagram of protein corona detected device using reflectometric interference spectroscopy based on silica colloidal crystal films

图4. (a)梯度浓度(0.8, 1.2, 1.6和2 mol/L)的盐酸溶液对二氧化硅胶体晶体薄膜实时光学厚度的变化的影响(ΔOT); (b) HCl浓度与光学厚度变化的线性关系

Figure 4. (a) Real-time changes in the optical thickness (ΔOT) of silica colloidal crystal films with perfused ultrapure water, 0.8, 1.2, 1.6 and 2 mol/L of HCl; (b) linear relationship between concentration of HCl and optical thickness variation

图5. (a)不同浓度(2, 1, 0.5, 0.1和0.05 mg/mL)的Fg在二氧化硅胶体晶体薄膜蛋白冠形成引起的实时光学厚度变化(ΔOT)实时变化图; (b) 6000 s时不同浓度Fg蛋白冠形成引起的ΔOT值

Figure 5. (a) Real-time ΔOT caused by different concentrations (2, 1, 0.5, 0.1 and 0.05 mg/mL) of Fg protein corona formation in silica colloidal crystal film; (b) ΔOT caused by different concentration Fg formation of protein corona value at time of 6000 s

图6. (a) HSA, (b) Fg和(c) IgG结构模型和在二氧化硅胶体晶体薄膜中蛋白冠示意图; (d) HSA, Fg和IgG在二氧化硅胶体晶体薄膜蛋白冠形成过程引起的实时光学厚度变化(ΔOT)

Figure 6. A schematic representation of structure and protein corona formation of (a) HSA, (b) Fg and (c) IgG; (d) IgG, HSA and Fg protein corona formation on silica colloidal crystal film and real-time optical thickness changes (ΔOT)

图7. HSA在110, 190和280 nm二氧化硅胶体晶体薄膜蛋白冠形成过程引起的实时光学厚度变化(ΔOT)

Figure 7. Real-time ΔOT of protein corona formation for HSA at 110, 190 and 280 nm silica colloidal crystal film

图8. 血浆全血样品在190 nm二氧化硅胶体晶体薄膜蛋白冠形成过程引起的实时光学厚度变化(ΔOT)

Figure 8. Real-time ΔOT of protein corona formation for plasma and whole blood sample at 190 nm silica colloidal crystal film

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