Biological and Soft Matter Physics, Huygens-Kamerlingh Onnes Laboratory, Leiden University , P.O. Box 9504, 2300 RA Leiden, The Netherlands.
Fraunhofer Institute for Silicate Research , Neunerplatz 2, 97082 Würzburg, Germany.
Langmuir. 2017 Sep 26;33(38):9803-9810. doi: 10.1021/acs.langmuir.7b02065. Epub 2017 Sep 13.
Colloidal particles with specific binding affinity are essential for in vivo and in vitro biosensing, targeted drug delivery, and micrometer-scale self-assembly. Key to these techniques are surface functionalizations that provide high affinities to specific target molecules. For stabilization in physiological environments, current particle coating methods rely on adsorbed surfactants. However, spontaneous desorption of these surfactants typically has an undesirable influence on lipid membranes. To address this issue and create particles for targeting molecules in lipid membranes, we present here a surfactant-free coating method that combines high binding affinity with stability at physiological conditions. After activating charge-stabilized polystyrene microparticles with EDC/Sulfo-NHS, we first coat the particles with a specific protein and subsequently covalently attach a dense layer of poly(ethyelene) glycol. This polymer layer provides colloidal stability at physiological conditions as well as antiadhesive properties, while the protein coating provides the specific affinity to the targeted molecule. We show that NeutrAvidin-functionalized particles bind specifically to biotinylated membranes and that Concanavalin A-functionalized particles bind specifically to the glycocortex of Dictyostelium discoideum cells. The affinity of the particles changes with protein density, which can be tuned during the coating procedure. The generic and surfactant-free coating method reported here transfers the high affinity and specificity of a protein onto colloidal polystyrene microparticles.
具有特定结合亲和力的胶体颗粒对于体内和体外生物传感、靶向药物输送和微米级自组装至关重要。这些技术的关键是表面功能化,它为特定靶分子提供高亲和力。为了在生理环境中稳定,目前的颗粒涂层方法依赖于吸附的表面活性剂。然而,这些表面活性剂的自发解吸通常对脂质膜有不良影响。为了解决这个问题并为靶向脂质膜中分子的颗粒创造条件,我们在此提出了一种无表面活性剂的涂层方法,该方法将高结合亲和力与生理条件下的稳定性结合在一起。在用 EDC/Sulfo-NHS 激活带电荷稳定的聚苯乙烯微球后,我们首先用特定的蛋白质涂层颗粒,然后共价连接一层密集的聚乙二醇。该聚合物层在生理条件下提供胶体稳定性和抗粘性,而蛋白质涂层提供对靶向分子的特异性亲和力。我们表明,链霉亲和素功能化的颗粒特异性地结合到生物素化的膜上,而伴刀豆球蛋白 A 功能化的颗粒特异性地结合到盘基网柄菌细胞的糖萼上。颗粒的亲和力随蛋白质密度而变化,这可以在涂层过程中进行调整。这里报道的通用且无表面活性剂的涂层方法将蛋白质的高亲和力和特异性转移到胶体聚苯乙烯微球上。