SBIR Phase II: Magnetic Quantum Dots for Cell Separation and Characterization

该SBIR二期项目的更广泛/商业影响将开发磁性和荧光试剂，用于分离和分析储存在生物库中的组织样本（包括组织、细胞、血液、血清和尿液等生物样本的中央存储库）及其相关患者数据中的细胞，用于研究确定诊断和治疗疾病的新方法；该行业目前的年复合增长率为4.5%，达到520亿美元。生物库样本通常由病理正常和病变细胞和组织的混合物组成，需要分离。因此，样本质量对生物银行至关重要。细胞分离市场（39亿美元/年，复合年增长率4.1%）由流式细胞术领域（31亿美元/年，复合年增长率3.6%）主导；而磁细胞分离是一个不断增长的市场（到2020年复合年增长率为6.4%，每年5.82亿美元）。大多数生物银行目前提供未纯化的样品或仅通过流式细胞术纯化的样品，在纯度为~20%的情况下，典型的回收率为10%。这项研究将开发和商业化结合磁分离和流式细胞术分析的试剂，用于从生物库组织样本中纯化靶细胞。在公司对悬浮细胞的一期研究中，这些试剂回收率为~100%，纯度为75%。这项研究将把这项技术扩展到组织匀浆中，增加不均匀性和粘度，提高生物库样品的纯度，增强研究人员开发突破性医疗技术的能力。本研究将开发磁荧光纳米粒子试剂，用于生物库中异质临床组织样本的细胞分离和随后的流式细胞术分析。鉴于生物标本的异质性，可能含有比患病成分更多的正常成分，纯化技术对生物库的操作至关重要。根据第一阶段的研究，该公司预计该技术可以将回收率和纯度提高到75%（目前为10-20%）。这将通过以下方式实现：(1)开发一种开放式磁分离系统，以减少与现有商用磁珠填充柱相比的阻塞；(2)优化试剂磁性和荧光含量，最大限度地提高细胞分离和分析信号，同时最大限度地减少大小和信号与组织自身荧光的重叠；(3)制定组织匀浆分析的方案，使用常用的机械和酶组织消化方法。这项研究将产生从复杂、粘性环境中分离细胞的新工具，这将大大提高生物库样本在研究和临床研究中的效用。这些改进也将有利于不太严格的工程设计要求的应用，包括生物/制药分离、生物处理（例如，CAR - t细胞治疗）和临床诊断（例如，循环肿瘤细胞恢复）。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader/commercial impact of this SBIR phase II project will develop magnetic and fluorescent reagents for the separation and analysis of cells from tissue samples stored in biobanks, central repositories for biological samples, including tissues, cells, blood, sera, and urine, and their associated patient data, used in research to identify new methods to diagnose and treat disease; this industry is currently $52 B/yr with a compound annual growth rate (CAGR) of 4.5%. Biobank samples typically consist of a mixture of pathologically normal and diseased cells and tissues, requiring separation. Sample quality is thus critical for biobanks. The cell separation market ($3.9 B/yr, CAGR 4.1%) is dominated by the flow cytometry segment ($3.1 B/yr, CAGR 3.6%); whereas magnetic cell separation is a growing market ($582 M/yr by 2020 CAGR of 6.4%). Most biobanks currently provide unpurified samples or samples purified via flow cytometry alone, with typical recoveries of 10% at purities of ~20%. This research will develop and commercialize reagents that combine magnetic separation with flow cytometry analysis for target cell purification from biobank tissue samples. In the company's Phase I research of cells in suspension, these reagents yielded ~100% recovery with purities 75%. This research will expand this technology to tissue homogenates with increased heterogeneity and viscosity, increasing purity of biobank samples and enhancing researcher ability to develop breakthrough medical technologies.This research will develop magnetic-fluorescent nanoparticle reagents for cell separation and subsequent flow cytometry analysis of heterogeneous clinical tissue samples from biobanks. Given the heterogeneous nature of biological specimens that may contain more normal than diseased components, purification technologies are critical to biobank operation. Based on the Phase I research, the company anticipates that this technology could increase recovery and purity to 75% (vs. 10-20% currently). This will be accomplished by: (1) developing an open magnetic separation system to reduce obstruction compared to existing commercial magnetic bead-packed columns; (2) optimization of reagent magnetic and fluorescent content to maximize cell separation and analysis signal while minimizing size and signal overlap with tissue autofluorescence; (3) protocol development for tissue homogenate analysis using commonly employed mechanical and enzymatic tissue digestion methods. This research will yield new tools for cell isolation from complex, viscous environments that will greatly enhance the utility of biobank samples in research and clinical investigations. Such improvements will also benefit applications with less rigorous engineering design requirements, including bio/pharma separations, bioprocessing (e.g., CAR T-cell therapy), and clinical diagnostics (e.g., circulating tumor cell recovery).This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.