Continuous exosome and oncosome separations using a modified SPLITT system

胞外体和胞外体是分别由正常细胞和癌细胞产生的胞外囊泡，存在于大多数体液中。由于肿瘤体与癌症转移密切相关，因此在临床和科学上具有越来越重要的意义。为了满足临床液体的高通量和大容量处理的挑战，提出了一种设备，通过从临床液体中连续分离内体/外体，然后从外体中分离内体和外体。该设备基于外体/内含体的物理化学差异(如大小、密度和电荷)而不是亲和力技术进行分离。场流分离将显示可行性，并发现黑色素瘤细胞和黑素细胞培养细胞的胞外体和胞外体之间存在的物理性质差异。血清将与几种分流侧向输送薄层分离(Splitt)仪器一起使用。Splitt将被用来分离外体和内体，并与标准的外体精制技术进行比较。在成功完成这些技术目标后，将使用详细的成本/时间分析来证明第二阶段资金中对Beta原型的开发、额外的临床样本和癌症类型的评估是合理的。

Both exosomes and oncosomes are extracellular vesicles produced by normal and cancerous cells respectively, and are found in most bodily fluids. Oncosomes are of increasing clinical and scientific importance because they are involved in cancer metastasis. A device is presented to meet the challenge of high throughput and high-volume processing of clinical fluids, by continuously separating oncosomes/exosomes from the clinical fluid, and then separating the oncosomes from the exosomes. The device separates based upon physicochemical differences (such as size, density, and charge) of the exosomes/oncosomes rather than upon affinity techniques. Field flow fractionation will show feasibility and discover the physical property differences that exist between oncosome and exosome from cultured cells of melanoma and melanocytes respectively. Serum will be used with several split-flow lateral-transport thin separation (SPLITT) instruments. SPLITT will be used to separate exosome from oncosome and compared against standard techniques for exosome refinement. Upon successful completion of these technical objectives, a detailed cost/time analysis will be used to justify the evaluation of development of beta prototype, additional clinical samples, and cancer types in Phase II funding.