Buoyancy based bioseparations

摘要 对高纯度分离稀有金属的高成本高效技术的需求尚未得到满足 血液中的循环肿瘤细胞(CTCs)。理想的隔离技术应该能够 满足以下标准：(A)处理时间最短；(B)最终样本量少；(C) 从大量血液中分离；(D)白细胞和血液的低残留 组件；(E)高细胞活力；(F)与下游工艺的兼容性，例如 聚合酶链式反应或组织培养；(G)成本低，入门级人员易于操作。 虽然有许多分离CTC的方法，但没有一种方法结合 简单、低成本、高效率、高速度。在此之前，我们开发了一种新的工艺 浮力微泡辅助从血液和棕黄色大衣中分离罕见的肿瘤细胞。 MBS高效(80%回收率)和快速(少于20分钟)分离稀有肿瘤细胞 从7毫升的血液或22毫升的巴菲大衣。对于第一阶段，诊断，有限责任公司已与 加州大学圣地亚哥分校为了开发和测试一种 直观、用户友好的分离/收获设备，与MBS相结合将 用于从复杂的生物液(如血液)中获取罕见的肿瘤细胞。这个 建议的设置将使洗涤、免疫染色、遗传分析或培养变得容易 孤立的CTC。MB方法与 磁珠将高速、样品纯度、简单性、灵活性、终止性降至最低 样品量大，成本低。这项技术的价值主张是定性的 以及生物医学和临床CTC分离和分析的定量改进 研究和潜在的临床诊断学。

Abstract There is an unmet need for cost-efficient technologies for high purity isolation of rare circulating tumor cells (CTCs) from blood. An ideal isolation technique should be able to meet the following criteria: (a) minimal processing time; (b) low final sample volume; (c) isolation from large blood volume; (d) low carryover of leukocytes and blood components; (e) high cell viability; (f) compatibility with downstream processes such as PCR or tissue culturing; (g) low cost and ease of operation by entry-level personnel. While there are numerous approaches for CTC isolation, none of them combines simplicity, low cost, efficiency and speed. Previously, we developed a novel process of buoyant microbubble-assisted isolation of rare tumor cells from blood and buffy coats. MBs efficiently (>80% recovery) and fast (less than 20 min) isolated rare tumor cells from 7 ml blood or 22 ml buffy coats. For Phase I, Diagnologix, LLC has partnered with the University of California San Diego in order to develop and test a prototype of an intuitive and user-friendly separation/harvesting device that in combination with MBs will be used for harvesting rare tumor cells from complex biological fluids (e.g., blood). The proposed setup will enable easy washing, immunostaining, genetic analysis or culturing of isolated CTCs. The most important competitive advantages of MB approach vs. magnetic beads would be high speed, sample purity, simplicity, flexibility, minimal final sample volume and low cost. The value proposition of the technology is in the qualitative and quantitative improvement in CTC isolation and analysis for biomedical and clinical research and potentially for clinical diagnostics.