Commercialization of Magnetic Nanobeads for Cancer Cell Enrichment

描述（由申请人提供）： 哥伦布纳米工程（CNW）提出的研究导致商业化的定制磁性纳米粒子试剂的检测癌细胞从各种来源，如骨髓和外周血。细胞分离对于从复杂细胞群中分离稀有细胞的研究人员和临床医生来说越来越普遍，以了解细胞在不同环境中的功能或诊断疾病的发生，复发或进展。复杂的混合物和寻找细胞样品中的微小变化需要新的技术，提供高通量的灵敏度;因此，细胞分离技术的进步必须允许研究人员找到低百分比的感兴趣细胞，并纯化它们以准确评估生物相关性。在第一阶段研究中，CNW证明了磁性纳米颗粒的制造，其尺寸均匀，在50至70 nm范围内，配体密度为2至5 ug/ml Fe颗粒，并引起免疫磁性标记细胞的磁泳迁移率比市售磁性试剂大一个数量级。在I期申请中实现的上述定义磁性颗粒的标准允许在107个总细胞中富集1个肿瘤细胞，平均回收率为74.1%的加标肿瘤样品，以及在称为四极磁分选仪（QMS）的流通磁分选仪中平均3.21 log 10的污染细胞消耗。因此，第二阶段的研究要求设计和商业化的磁性试剂盒，具体用于QMS应用涉及癌细胞富集。CNW将（1）扩大处理工作，以获得比当前处理技术增加50倍的效果;（2）建立程序和实践，确保对纳米颗粒大小和磁性的一致控制;（3）开发用于商业磁流分选机QMS的CNW细胞富集试剂盒的方案;以及（4）使用乳腺癌患者和头颈癌患者进行方案的beta测试。拟定的第II阶段最终产品是专门针对QMS系统的定制磁性试剂盒，可从各种血液来源（如骨髓和外周血）中富集癌细胞。颗粒的质量标准将提供大于70%的所需细胞回收率，以及4 log 10的污染细胞消除。在QMS系统中使用CNW纳米颗粒的所得富集级分将提供质量纯化的产物用于进一步的下游处理和随后的分子分析。我们的目标是使CNW定制试剂成为当前病理学库中金标准检测的组成部分。哥伦布NanoWorks是一家磁性纳米颗粒试剂公司，提供磁性细胞分离试剂。在本申请中，我们将开发磁性试剂盒作为四极磁分选（QMS）系统的商业试剂。在过去，QMS系统中使用的商业磁性纳米颗粒在所需细胞的纯度和回收方面提供了低于标准的结果。哥伦布NanoWorks在其I期申请中证明，为QMS定制设计的试剂能够实现平均3.21 log 10的污染细胞消除，以及从对照样品中平均回收74.1%的肿瘤细胞。我们相信，CNW为QMS系统定制的试剂盒将提供更可预测的、可重复的结果。

DESCRIPTION (provided by applicant): Columbus NanoWorks (CNW) proposes research leading to the commercialization of custom magnetic nanoparticle reagents for the detection of cancer cells from various sources such as bone marrow and peripheral blood. Cell separation is becoming increasingly commonplace for researchers and clinicians who isolate rare cells from complex cell populations to understand how cells function in different environments or to diagnose disease occurrence, recurrence or progression. Complex mixtures and searches for small variations in cell samples require new techniques that provide exquisite sensitivity with high throughput; therefore, advances in cell separation technology must allow the researcher to find low percentage cells of interest, as well as purify them to allow for accurate assessment of biological relevance. In Phase I research CNW demonstrated the manufacture of magnetic nanoparticles that were uniform in size within a range of 50 to 70nm, had a ligand density of 2 to 5 ug/ml Fe particles and elicited a magnetophoretic mobility of immunomagnetically labeled cells an order of magnitude greater than commercially available magnetic reagents. The aforementioned criteria defining magnetic particles achieved in the Phase I application, allowed the enrichment of 1 tumor cell in 107 total cells with an average recovery of 74.1% of spiked tumor samples, as well as an average 3.21 log10 depletion of contaminating cells in the flow-through magnetic sorter known as the Quadrupole Magnetic Sorter (QMS). Phase II research thus calls for the design and commercialization of magnetic reagent kits specific for QMS applications involving cancer cell enrichment. CNW will (1) scale up processing efforts to obtain a 50 fold increase over current process techniques; (2) establish procedures and practices which assure consistent control of nanoparticle size and magnetic properties; (3) develop protocols for CNW cell enrichment kits for use in the commercial magnetic flow sorter, QMS; and (4) beta testing of protocols using breast cancer patients and head and neck cancer patients. The proposed Phase II end items are customized magnetic reagent kits specific for the QMS system that will allow the enrichment of cancer cells from various blood sources such as bone marrow and peripheral blood. Specifications of the particles will provide a greater than 70% recovery of desired cells, as well as a 4 log10depletion of contaminating cells. The resultant enriched fraction using CNW nanoparticles in the QMS system will provide a quality-purified product for further downstream processing and subsequent molecular analysis. The goal is for CNW customized reagents to become an integral part of gold standard testing in the current pathology repertoire. Columbus NanoWorks is a magnetic nanoparticle reagent company that provides reagents for magnetic cell separation. In this application we will develop magnetic reagent kits as commercial reagents for the Quadrupole Magnetic Sorter (QMS) system. In the past, commercial magnetic nanoparticles used in the QMS system have provided sub-par results with respect to purity and recovery of desired cells. Columbus NanoWorks demonstrated in their Phase I application that reagents custom designed for the QMS were able to achieve an average 3.21 log10 depletion of contaminating cells as well as an average recovery of 74.1% of tumor cells from control samples. We believe that CNW's custom reagent kits for the QMS system will provide more predictable, reproducible results from sample to sample.