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首页> 外文期刊>Acta biomaterialia >Avidin-conjugated calcium phosphate nanoparticles as a modular targeting system for the attachment of biotinylated molecules in vitro and in vivo
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Avidin-conjugated calcium phosphate nanoparticles as a modular targeting system for the attachment of biotinylated molecules in vitro and in vivo

机译：抗生物素蛋白缀合的磷酸钙纳米粒子作为用于在体外和体内附着生物素化分子的模块化靶向系统

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Avidin was covalently conjugated to the surface of calcium phosphate nanoparticles, coated with a thin silica shell and terminated by sulfhydryl groups (diameter of the solid core about 50 nm), with a bifunctional crosslinker connecting the amino groups of avidin to the sulfhydryl group on the nanoparticle surface. This led to a versatile nanoparticle system where all kinds of biotinylated (bio-)molecules can be easily attached to the surface by the non-covalent avidin-biotin-complex formation. It also permits the attachment of different biomolecules on the same nanoparticle (heteroavidity), creating a modular system for specific applications in medicine and biology. The variability of the binding to the nanoparticle surface of the was demonstrated with various biotinylated molecules, i.e. fluorescent dyes and antibodies. The accessibility of the conjugated avidin was demonstrated by a fluorescence-quenching assay. About 2.6 binding sites for biotin were accessible on each avidin tetramer. Together with a number of about 240 avidin tetramer units per nanoparticle, this offers about 600 binding sites for biotin on each nanoparticle. The uptake of fluorescently labelled avidin-conjugated calcium phosphate nanoparticles by HeLa cells showed the co-localization of fluorescent avidin and fluorescent biotin, indicating the stability of the complex under cell culture conditions. CD11c-antibody functionalized nanoparticles specifically targeted antigen-presenting immune cells (dendritic cells; DCs) in vitro and in vivo (mice) with high efficiency.
机译：伊霉素与磷酸钙纳米粒子的表面共价缀合，涂有薄二氧化硅壳，并由巯基（固体核心的直径约为50nm）终止，其中双官能交联剂将伊霉素的氨基连接到巯基上的氨基纳米粒子表面。这导致了通过非共价抗生物素蛋白 - 生物素 - 复杂的形成容易地连接到表面的多功能纳米颗粒系统，其中可以通过非共价抗生物素 - 生物素复合物形成。它还允许在同一个纳米颗粒（杂差）上附着不同的生物分子，为医学和生物学中的特定应用产生模块化系统。用各种生物素化分子对纳米颗粒表面进行结合的可变性，即荧光染料和抗体。通过荧光猝灭测定证明了共轭抗杀虫蛋白的可访问性。在每次抗生物素蛋白四聚体上可获得大约2.6个用于生物素的结合位点。每纳米颗粒的许多约240个抗生物素蛋白四聚体单元，这为每种纳米颗粒上的生物素提供约600个结合位点。通过HeLa细胞吸收荧光标记的抗磷酸磷酸钙纳米粒子，荧光抗生物素蛋白和荧光生物素的共定位，表明在细胞培养条件下复合物的稳定性。 CD11C-抗体官能化纳米颗粒特异性地靶向抗原呈现免疫细胞（树突状细胞; DCS），高效，体内（小鼠）。

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