Department of Veterinary Medicine, National Taiwan University . 1 Sec. 4, Roosevelt Road, Taipei 10617, Taiwan.
Research Center for Nanotechnology and Infectious Diseases , Taipei, Taiwan.
ACS Appl Mater Interfaces. 2017 Nov 22;9(46):39953-39961. doi: 10.1021/acsami.7b09931. Epub 2017 Nov 8.
Attachment to cellular surfaces is a major attribute among infectious pathogens for initiating disease pathogenesis. In viral infections, viruses exploit receptor-ligand interactions to latch onto cellular exterior prior to subsequent entry and invasion. In light of the selective binding affinity between viral pathogens and cells, nanoparticles cloaked in cellular membranes are herein employed for virus targeting. Using the influenza virus as a model, erythrocyte membrane cloaked nanoparticles are prepared and modified with magnetic functionalities (RBC-mNP) for virus targeting and isolation. To maximize targeting and isolation efficiency, density gradient centrifugation and nanoparticle tracking analysis were applied to minimize the presence of uncoated particles and membrane vesicles. The resulting nanoparticles possess a distinctive membrane corona, a sialylated surface, and form colloidally stable clusters with influenza viruses. Magnetic functionality is bestowed to the nanoparticles through encapsulation of superparamagnetic iron-oxide particles, which enable influenza virus enrichment via magnetic extraction. Viral samples enriched by the RBC-mNPs result in significantly enhanced virus detection by multiple virus quantification methods, including qRT-PCR, immunnochromatographic strip test, and cell-based titering assays. The demonstration of pathogen targeting and isolation by RBC-mNPs highlights a biologically inspired approach toward improved treatment and diagnosis against infectious disease threats. The work also sheds light on the efficient membrane cloaking mechanism that bestows nanoparticles with cell mimicking functionalities.
细胞表面附着是传染性病原体引发疾病发病机制的主要特性。在病毒感染中,病毒利用受体-配体相互作用,在随后的进入和入侵之前附着在细胞外部。鉴于病毒病原体与细胞之间的选择性结合亲和力,本文采用细胞膜包裹的纳米粒子进行病毒靶向。以流感病毒为模型,制备了红细胞膜包裹的纳米粒子,并修饰了磁性(RBC-mNP)以进行病毒靶向和分离。为了最大限度地提高靶向和分离效率,应用密度梯度离心和纳米粒子跟踪分析来最小化未涂层颗粒和膜泡的存在。所得纳米粒子具有独特的膜冠,表面具有唾液酸化,并与流感病毒形成胶体稳定的聚集体。通过超顺磁性氧化铁颗粒的封装为纳米粒子赋予了磁性功能,这使得可以通过磁性提取来富集流感病毒。通过 RBC-mNP 富集的病毒样本通过多种病毒定量方法,包括 qRT-PCR、免疫层析条带试验和基于细胞的滴度测定法,显著提高了病毒检测的灵敏度。RBC-mNP 对病原体的靶向和分离的证明突出了一种受生物启发的方法,可改善针对传染病威胁的治疗和诊断。这项工作还揭示了赋予纳米粒子细胞模拟功能的高效膜包裹机制。