Characterization of Biological Nanoparticle Subsets

生物纳米颗粒子集的表征

• 批准号: 10014708
• 负责人: Jennifer Jones
• 金额: $ 152.27万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10014708
• 关键词: B-Lymphocytes; Biological; Biological Markers; Biology; Blood; Calibration; Cells; Clinical; Data; Detection; Dimensions; Epitopes; FOLH1 gene; Flow Cytometry; Fluorescence; Goals; Hypoxia; Immune; Immune response; Immunobiology; Immunotherapy; Individual; Label; Laboratories; Leukocytes; Malignant Neoplasms; Methods; MicroRNAs; Molecular; Monitor; Monoclonal Antibodies; Plasma; Population; Proteins; Protocols documentation; RNA; Radiation; Reporting; Research; Resolution; Robotics; Sorting - Cell Movement; Source; Surveys; T-Lymphocyte; TACSTD1 gene; TGFB1 gene; Technology; Tumor Biology; Tumor stage; Vaccines; Vesicle; analysis pipeline; base; burden of illness; cancer therapy; exosome; extracellular vesicles; high dimensionality; improved; instrumentation; nanoparticle; neoplastic cell; novel marker; particle; programs; receptor; response; single molecule; stressor; therapeutic target; treatment response; tumor; tumor progression

During the past year, we have developed a comprehensive pipeline for EV analysis, which we refer to as the Multiplex-to-Single-EV-Analysis (Mt-SEA) pipeline. This Mt-SEA pipeline includes multiplex high dimensional EV surveys as well as high resolution single EV studies, including nanoFACS protocols for fluorescence detection and calibration, for counting, and for high-throughput sorting, to sort EV subsets from plasma and other biofluids by nanoFACS. Using multiple distinct vesicle populations, we find that nanoFACS sorting is uniquely able to produce high fidelity EV subsets (95-97% pure), with functional biological cargo (RNA and protein). Our current focus is exclusively on application of this method for clinically important studies, for the study of cancer disease burden, metastatic potential, and responses to treatment. Furthermore, we have devised a new class of labels (Molecular NanoTags) that can be used with nanoFACS, not only to improve the detection of EV epitopes, but also to enable single molecule counting by flow cytometry. The specialized instrumentation, expertise, and biospecimen pipeline that we have assembled and are continuing to improve for this nanoFACS program is a unique and distinctive asset for NCI.

在过去的一年中，我们开发了一条综合的EV分析管道，我们称之为多路复用到单个-EV分析（MT-SEA）管道。该MT-SEA管道包括多重高维EV调查以及高分辨率的单EV研究，包括用于荧光检测和校准的纳米FACS协议，用于计数以及用于高通量分类，以从血浆和其他纳米氟酸酯中对EV子集进行分类。使用多个不同的囊泡种群，我们发现纳米型分类具有独特的能力，能够产生高富达EV子集（95-97％纯），具有功能性生物货物（RNA和蛋白质）。我们目前的重点是将这种方法应用于临床上重要的研究，用于研究癌症疾病负担，转移性潜力和对治疗的反应。此外，我们设计了一类新的标签（分子纳米塔），可与纳米量一起使用，不仅可以改善对EV表位的检测，而且还可以通过流式细胞仪进行单分子计数。我们已经组装并继续改进该纳米型计划的专门仪器，专业知识和生物循环管道是NCI的独特而独特的资产。