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

为了提供一个适合于提纯肿瘤来源的小泡并将其与生物液中的其他组织来源的小泡分离的平台，我们提出了一种两步法：1)Aethlon Medical的血液净化器，用于从血浆等生物体液中分离出总的外周体群。这种亲和生物过滤技术采用中空纤维血液滤芯(平均孔径200 nm)，内含凝集素雪花甘露凝集素(GNA)作为亲和剂，已被用于捕获大小相似的广泛范围的病毒和高甘露糖糖蛋白表面的外体。这种临床使用的设备已经缩小到能够过滤大量复杂生物液的台式版本，从而克服了当前外体分离技术的局限性。2)用识别肿瘤外切体上表达的硫酸软骨素蛋白多糖4表位的单抗225.28磁性免疫捕获肿瘤外切体。共同研究人员怀特赛德博士和费龙博士已经证明了使用单抗225.28对黑色素瘤外显体进行磁分离的能力。通过利用专门用于快速处理生物液的设备平台，然后用单抗225.28免疫捕获，拟议的工作旨在标准化一种从复杂生物液中分离外体的方法，该方法将适用于研究和临床护理的自动化工作流程。

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.