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.

项目摘要/摘要 希望将基础细胞生物学、疾病传播机制和临床联系起来的愿望 诊断学是新技术发展的主要驱动力。在可能的生物物种中 如果沿着这些路线开发，外显色体尤其有希望。外切体很小(30-130 nm) 来自体内所有细胞类型的细胞外小泡(EV)，现在被实现为 细胞内通讯。它们展示了它们起源于细胞的蛋白质生物标记物，使它们变得很有前途 用于疾病诊断的候选材料。同样，人们对外切体的潜在用途也很感兴趣 药物运载工具(即病媒)。然而，在经济衰退之前，有必要进行大量的基础研究。 充分实现了外切体的用途。基于外显子的研究和应用中的一个关键挑战 应用在于缺乏健壮和通用的方法从不同的生物介质中分离囊泡。 虽然分离和量化方法已经发展，但没有一种方法能够克服与 严密性和重复性，以干净、快速和经济有效的方式分离外切体。为了填补这一空白，一个 极高效的外切体捕获和分离平台技术，基于新型的聚乙烯- (对苯二甲酸)(PET)毛细通道聚合物(C-CP)纤维和薄膜正在开发应用 在与基础研究、临床诊断和准备恢复相关的范围内。初步结果 结果表明，纤维能有效地分离胞外囊泡，富含外周小体，大小分布均匀 产量可与传统分离方法相媲美，时间短得多，体积小， 更高的纯度。建议进一步发展和验证这种外显子分离方法。 用于基础研究和临床实验室，并扩展到载体的准备规模 申请。按照本计划目标的要求和集体“外显体”的要求 社区“，努力的最终目标是交付工作原型，以供科学评估 同行和潜在的商业提供商。研究和开发计划分三个阶段进行 明确的目标。在第一个，由独特形状的纤维制成的高渗透性色谱柱 为分离纯化适于临床、研究和应用的外切体提供了平台 和制备规模，优于目前的外切体分离方法。第二，实施 向下旋转尖端形式的纤维为普通和靶向外切体提供了更多的通用性 使用普通台式离心机进行收获。在第三种情况下，C-CP纤维膜可以被配置为影响 用于临床诊断的高效、多重侧向流动免疫分析。我们完全相信，这些结果 将证明新型C-CP光纤/薄膜隔离平台将被证明是一种高效、低成本的 分离外切体的有效手段，用于基础生化研究、临床诊断和 预备性应用，这些特点将导致该平台的商业可用性。