IGF::OT::IGF Phase I SBIR Topic 359 Title: Device Strategy for Selective Isolation of Oncosomes and Non-Malignant Exosomes09/15/2017 - 06/14/2018Firm Fixed Price

IGF::OT::IGF 第一阶段 SBIR 主题 359 标题：选择性分离肿瘤体和非恶性外泌体的设备策略09/15/2017 - 06/14/2018固定价格

• 批准号: 9570538
• 负责人: Annette Marleau
• 金额: $ 29.93万
• 依托单位: AETHLON MEDICAL, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2018-06-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9570538
• 关键词: Affinity; Agglutinins; Chondroitin Sulfate Proteoglycan; Clinical; Complex; Devices; Epitopes; Fiber; Galanthus nivalis; Hemofiltration; Lectin; Magnetism; Mannose; Medical; Membrane Glycoproteins; Methods; Non-Malignant; Phase; Plasma; Population; Price; Recovery; Research; Research Personnel; Small Business Innovation Research Grant; Standardization; Techniques; Technology; Tissues; Tumor-Derived; Vesicle; Virus; Work; base; clinical care; exosome; melanoma; tumor

To provide a platform suitable for purification of tumor-derived vesicles and to separate them from other tissue-derived vesicles in biofluids, we propose a two-step approach: 1) Aethlon Medical’s Hemopurifier device for isolation of the total exosome population from biofluids such as plasma. This affinity biofiltration technology employs hollow-fiber hemofiltration cartridges (average 200 nm pores) containing the lectin Galanthus nivalis agglutinin (GNA) as an affinity agent, which has been leveraged for capture of a broad spectrum of viruses of similar sizes and high mannose-glycoprotein surfaces as exosomes. This clinically-used device has been scaled down to a bench-top version capable of filtering large volumes of complex biofluids, thereby overcoming limitations of current exosome isolation techniques. 2) Magnetic immunocapture of tumor exosomes using the mAb 225.28 recognizing an epitope of chondroitin sulfate proteoglycan 4 expressed on tumor exosomes. The Co-Investigators, Drs. Whiteside and Ferrone, have demonstrated the ability to perform magnetic isolation of melanoma exosomes using mAb 225.28. By leveraging a device platform specialized for rapid processing biofluids followed by immunocapture with mAb 225.28, the proposed work aims to standardize an exosome isolation method from complex biofluids that will be suitable for automated workflows in research and clinical care.

为了提供适合于纯化肿瘤衍生的囊泡并将其与生物流体中的其他组织衍生的囊泡分离的平台，我们提出了两步方法：1）Aethlon Medical的Hemopurifier装置，用于从生物流体如血浆中分离总外泌体群体。这种亲和生物过滤技术采用中空纤维血液过滤滤芯（平均200 nm孔），含有凝集素雪花莲凝集素（GNA）作为亲和剂，已被用于捕获与外来体相似大小和高甘露糖糖蛋白表面的广谱病毒。这种临床使用的装置已经缩小到能够过滤大量复杂生物流体的台式版本，从而克服了当前外泌体分离技术的局限性。2)使用识别在肿瘤外泌体上表达的硫酸软骨素蛋白聚糖4的表位的mAb 225.28的肿瘤外泌体的磁性免疫捕获。共同研究者Whiteside和Ferrone博士已经证明了使用mAb 225.28进行黑色素瘤外泌体磁性分离的能力。通过利用专门用于快速处理生物流体的设备平台，然后用mAb 225.28进行免疫捕获，拟议的工作旨在标准化复杂生物流体中的外泌体分离方法，该方法将适用于研究和临床护理中的自动化工作流程。