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: 10796436
• 负责人: Terri Lane Foster Bruce
• 金额: $ 21万
• 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/10796436
• 关键词: Address; Affect; Basic Science; Biochemistry; Biological; Biological Process; Blood capillaries; Cells; Cellular biology; Characteristics; Clinical; Column Chromatography; Communication; Communities; Development; Disease; Drug Delivery Systems; Elements; Ethylenes; Exhibits; Fiber; Film; Harvest; Immunoassay; Laboratories; Lateral; Methodology; Methods; Permeability; Polymers; Positron-Emission Tomography; Program Development; Provider; Recovery; Reproducibility; Research; Scientific Evaluation; Shapes; Source; Therapeutic; Validation; Vesicle; cell type; clinical diagnostics; cost; cost effective; delivery vehicle; disease diagnostic; exosome; extracellular vesicles; fundamental research; intercellular communication; interest; knowledge base; new technology; novel; peer; programs; protein biomarkers; prototype; research and development; technology development; technology platform; terephthalate; 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.

项目概要/摘要 希望将基础细胞生物学、疾病传播机制和临床的各个方面联系起来 诊断是生物物种新技术发展的主要驱动力。 沿着这些路线进行开发，外泌体特别有前途。 细胞外囊泡 (EV) 源自体内所有细胞类型，现已被认为是 它们表现出来自其起源细胞的蛋白质生物标志物，这使得它们很有前途。 同样，人们对外泌体作为疾病诊断的潜在用途也很感兴趣。 然而，在药物输送载体（即载体）出现之前，还需要进行大量的基础研究。 外泌体的实用性是基于外泌体的研究和应用中的一个关键挑战。 应用在于缺乏从不同生物介质中分离囊泡的稳健且通用的方法。 尽管分离和定量方法不断发展，但没有一种方法能够克服与 清洁、快速且经济高效地分离外泌体的严格性和可重复性为了解决这一空白。 基于新型聚（乙烯）的极其高效的外泌体捕获和分离平台技术 (PET) 毛细管通道聚合物 (C-CP) 纤维和薄膜，正在开发用于应用 涵盖基础研究、临床诊断和预备性恢复的相关范围。 表明纤维可以有效地分离富含外泌体的细胞外囊泡，并且具有尺寸分布 其产量与传统分离方法相当，时间更短，体积规模更小，并且 这里建议进一步开发和验证这种外泌体分离方法。 用于基础研究和临床实验室，并扩展到载体制备规模 根据该计划的目标以及集体“外泌体”的要求。 社区”，努力的最终目标是提供工作原型以供科学评估 同行和潜在的商业供应商的研究和开发计划是在三个方面进行的。 具体目标 第一个是由独特形状的纤维制成的高渗透性色谱柱。 提供一个分离和纯化外泌体的平台，适合临床、研究、 和制备规模，优于当前的外泌体分离方法。 旋转尖端形式的纤维为通用和靶向外泌体提供了更大的多功能性 在第三种中，C-CP 纤维膜可以配置为影响 高效、多重侧流免疫分析的临床诊断结果令人充分相信。 该计划将证明新型 C-CP 纤维/薄膜隔离平台将被证明是一种高效、成本低廉的方法。 分离外泌体用于基础生物化学研究、临床诊断和 准备性应用程序，这些特征将导致平台的商业可用性。