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

A Novel Protein Corona Characterization based on the Reflectometric Interference Spectroscopy with Silica Colloidal Crystal Films

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.

It will form protein corona on the surface of nanoparticles when exposed to the biological environment.The pro-cess 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 etha-nol 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 im-munoglobulin 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.