A Scalable Continuous Production Platform for Large-Scale Manufacturing of Therapeutic Exosomes

用于大规模生产治疗性外泌体的可扩展连续生产平台

基本信息

批准号：
10739425
负责人：
Yichun Wang
金额：
$ 40.24万
依托单位：
UNIVERSITY OF NOTRE DAME
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-07 至 2025-08-31
项目状态：
未结题

项目摘要

Project Summary Exosomes are a subset of extracellular vesicles, with diameters between 30 and 200 nm, secreted by most eukaryotic cells. They deliver bioactive cargos, such as lipids, proteins and nucleic acids, to other target cells. Tremendous effort has been made recently on developing highly specific exosome-based drug delivery systems. Yet, there remain fundamental challenges: 1) current technologies for exosome enrichment and isolation cannot achieve sufficient throughput or purity; 2) drug loading efficiency into exosomes is very limited; and 3) the production of exosomes has yet to reach sufficient high throughput for clinical tests or even further development. To address these challenges, the proposed project utilizes a three-dimensional (3D) piezoelectric scaffold (PES) to stimulate exosome production, a lipid- membrane penetrating chiral graphene quantum dots (GQD) for efficiently load drugs into exosomes (>70% yield) and a scalable tangential-flow ultrafiltration module with ion-track asymmetric nanopore membranes (ANM) to isolate exosomes with high yield (>85%), throughput (240 mL/hour per module) and purity. The integration of three modules constitutes a high-throughput, high-purity, and scalable integrated platform for continuous manufacturing of therapeutic exosomes. The project is based on PI Wang’s preliminary experimental and computational results that GQDs facilitated drug loading into exosomes with higher efficiency than any current method, her extensive experience on nanocomposite for 3D tissue cultures, the co-PI Chang’s ANM exosome isolation technology for liquid biopsy and his patented technology on high-throughput AC nanofiber electrospinning. We will achieve the goal by following specific aims: 1) Develop high-throughput exosome production bioreactor module with stimulating PES. 2) Develop high-efficiency exosome drug loading module with chiral GQDs. 3) Develop and establish continuous ANM isolation and purification module. The proposed project contains several innovated approaches of exosome production, loading and isolation, if successful and integrated, can enhance therapeutic exosome production rate over current design, and achieve continuous production of therapeutic exosomes. Such an integrated production pipeline provides an innovative high-throughput and high-purity exosome manufacturing platform for drug delivery. They will also be designed to allow easy scale up for reactors of arbitrary size or for effluents from multiple reactors. Scaling to higher throughput can be achieved by just adding more parallel lines of the same modules.

项目概要外泌体是细胞外囊泡的一个子集，直径在 30 至 200 nm 之间，由大多数真核细胞将生物活性物质（例如脂质、蛋白质和核酸）传递给其他细胞。最近在开发高度特异性的基于外泌体的靶细胞方面做出了巨大的努力。然而，仍然存在根本性的挑战：1）当前的外泌体技术。富集和分离无法达到足够的通量或纯度；2）药物装载效率外泌体非常有限；3）外泌体的生产尚未达到足够高的通量为了解决这些挑战，拟议的项目利用了临床测试甚至进一步的开发。三维（3D）压电支架（PES）刺激外泌体产生，脂质穿膜手性石墨烯量子点（GQD）可有效地将药物装载到外泌体中（>70% 产率）和具有离子径迹不对称纳米孔的可扩展切向流超滤模块膜 (ANM) 分离外泌体，产量高 (>85%)，通量高（每个模块 240 mL/小时）三个模块的集成构成了高通量、高纯度、可扩展的系统。用于连续生产治疗性外泌体的集成平台该项目基于 PI。 Wang的初步实验和计算结果表明GQDs促进了药物负载外泌体的效率比任何现有方法都高，她在纳米复合材料方面的丰富经验对于 3D 组织培养，联合 PI Chang 的用于液体活检的 ANM 外泌体分离技术及其我们将通过高通量AC纳米纤维静电纺丝专利技术来实现这一目标。具体目标如下： 1）开发高通量外泌体生产生物反应器模块 2) 开发具有手性 GQD 的高效外泌体载药模块。开发并建立连续ANM分离纯化模块拟建项目。包含几种外泌体生产、装载和分离的创新方法，如果成功并且集成，可以比当前设计提高治疗性外泌体生产率，并实现这种集成的生产管道提供了治疗性外泌体的连续生产。用于药物输送的创新高通量和高纯度外泌体制造平台。还可以设计成可以轻松地放大任意尺寸的反应器或来自多个反应器的流出物只需添加更多相同的并行线即可实现更高吞吐量的扩展。模块。