Capillary-channeled polymers fibers and films - A platform technology for exosome isolation and analytics

毛细管通道聚合物纤维和薄膜 - 外泌体分离和分析的平台技术

• 批准号: 10419095
• 负责人: Terri Lane Foster Bruce
• 金额: $ 30.83万
• 依托单位: CLEMSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10419095
• 关键词: 3D Print; Address; Affect; Aliquot; Antibodies; Basic Science; Benchmarking; Biochemistry; Biological; Biological Assay; Biological Markers; Biological Process; Blood capillaries; Caliber; Cell Line; Cells; Cellular biology; Characteristics; Chemicals; Clinical; Clinical Research; Column Chromatography; Communication; Communities; Complex; Coupled; Culture Media; Development; Diagnosis; Diagnostic tests; Disease; Drug Delivery Systems; Elements; Environment; Ethylenes; Exhibits; Fiber; Film; Harvest; High Pressure Liquid Chromatography; Home; Image; Immobilization; Immunoassay; Laboratories; Lateral; Liquid substance; Membrane Proteins; Methodology; Methods; Modeling; Performance; Permeability; Phase; Plasma Cells; Polymers; Population; Preparation; Program Development; Proteins; Proteomics; Provider; Rapid diagnostics; Recovery; Reproducibility; Research; Sampling; Scientific Evaluation; Source; Surface; System; Technology; Therapeutic; Time; Urine; Validation; Vesicle; Virion; Western Blotting; aptamer; base; cell type; clinical diagnosis; clinical diagnostics; cost; cost effective; delivery vehicle; design; disease diagnostic; exosome; experimental study; extracellular vesicles; fluid flow; fundamental research; instrumentation; intercellular communication; interest; knowledge base; nanovesicle; new technology; novel; parallel processing; peer; programs; protein biomarkers; prototype; research and development; technology development; terephthalate; transcriptome sequencing; vector

Project Summary/Abstract The desire to tie aspects of fundamental cell biology, the mechanisms of disease propagation, and clinical diagnostics is a major driver for the development of new technologies. Among the biological species that may be exploited along these lines, exosomes are particularly promising. Exosomes are small (30–130 nm) extracellular vesicles (EVs) derived from all cell types within the body, which are now realized as key agents in intracellular communication. They exhibit protein biomarkers from their cells of origin, making them promising candidates for use in disease diagnostics. Likewise, much interest lies in the potential use of exosomes as drug delivery vehicles (i.e., vectors). However, a great deal of fundamental research is necessary before the utility of exosomes is fully realized. A crucial challenge in the application of exosome-based research and application lies in the lack of robust and versatile methods for vesicle isolation from diverse biological media. While isolation and quantification methods have evolved, none have overcome the key issues associated with rigor and reproducibility to cleanly, quickly, and cost-effectively isolate exosomes. To address this void, an extremely efficient platform technology for exosome capture and isolation, based on novel poly(ethylene- terephthalate) (PET) capillary-channeled polymer (C-CP) fibers and films, is being developed for applications across the scales of relevance for basic research, clinical diagnostics, and preparative recovery. Initial results show that the fibers can effectively isolate extracellular vesicles, enriched in exosomes, with size distributions and yields comparable to traditional isolation methods, in much shorter times, smaller volume scales, and higher purity. Proposed here is the further development and validation of this exosome isolation methodology for fundamental research and clinical laboratories, with extension to the preparative-scale for vector applications. As dictated by the objectives of this program, and as demanded by the collective “exosome community”, the ultimate objective of the effort is the delivery of working prototypes for evaluation by scientific peers and potential commercial providers. The Research and Development program is pursued across three Specific Aims. In the first, highly permeable chromatography columns created from unique-shaped fibers provide a platform for isolation and purification of exosomes amenable to applications on the clinical, research, and preparative scales, superior to current exosome isolation methodologies. In the second, implementation of the fibers in spin-down tip format provides greater versatility towards generic- and targeted-exosome harvesting using common, bench-top centrifuges. In the third, C-CP fiber films can be configured to affect a high-efficiency, multiplexed lateral flow immunoassay for clinical diagnostics. It is fully believed that the results of this program will demonstrate that novel C-CP fiber/film isolation platforms will prove to be an efficient, cost- effective means to isolate exosomes for use in fundamental biochemistry research, clinical diagnostics, and preparative applications, and that those characteristics will lead to commercial availability of the platfroms.

项目摘要/摘要 渴望将基本细胞生物学的各个方面，疾病传播的机制和临床 诊断是开发新技术的主要驱动力。在可能的生物物种中 沿着这些线被利用，外泌体特别有前途。外泌体很小（30-130 nm） 源自体内所有细胞类型的细胞外蔬菜（EV），现在已将其视为关键药物 细胞内通信。他们从原产地表现出蛋白质生物标志物，使它们承诺 用于疾病诊断的候选人。同样，很大的兴趣在于外泌体的潜在使用 药物输送车辆（即向量）。但是，必须在 外泌体的效用已完全实现。在应用基于外部的研究和 应用在于缺乏从潜水员生物学媒体中隔离蔬菜的强大和多功能方法。 虽然隔离和量化方法已经发展，但没有人克服与之相关的关键问题 严格且可重复的性能清洁，快速和成本效益分离外泌体。为了解决这个空白， 基于新型聚（乙烯 - 邻苯二甲酸酯（PET）毛细管通道聚合物（C-CP）纤维和膜正在用于应用 跨基础研究，临床诊断和准备恢复的相关性。初始结果 表明纤维可以有效地隔离富含外泌体的细胞外蔬菜，并具有尺寸分布 并产生与传统隔离方法相当的，在短时间内，较小的体积尺度和 较高的纯度。这里提出的是这种外泌体隔离方法的进一步发展和验证 用于基础研究和临床实验室，扩展到矢量准备的尺度 申请。正如该计划的目标所决定的，以及集体“外部”的要求 社区”，努力的最终目标是提供工作原型以评估科学 同行和潜在的商业提供商。研究与发展计划是在三个 具体目标。在第一个由独特形纤维创建的高度渗透色谱柱中 提供一个平台，用于隔离和纯化外泌体可与临床，研究，研究， 和准备的量表，优于当前外泌体分离方法。第二，实施 旋转尖端格式的纤维为通用和目标外观提供了更大的多功能性 使用常见的台式离心机收获。在第三个中，可以将C-CP纤维膜配置为影响 高效，多路复用的侧向流免疫测定法，用于临床诊断。完全相信结果 该程序将证明新型的C-CP光纤/膜隔离平台将被证明是一个有效的，成本 - 分离外泌体的有效方法用于基本生物化学研究，临床诊断和 制备应用，这些特征将导致平台的商业可用性。