Commercialization of Magnetic Nanobeads for Cancer Cell Enrichment

用于富集癌细胞的磁性纳米珠的商业化

• 批准号: 7941580
• 负责人: Arfaan Rampersaud
• 金额: $ 20万
• 依托单位: COLUMBUS NANOWORKS, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7941580
• 关键词: Benchmarking; Binding; Biological; Biological Assay; Biotin; Blood; Blood Cells; Bone Marrow; Buffers; Cancer Patient; Carbon; Cell Separation; Cell Survival; Cell physiology; Cells; Chemicals; Clinic; Clinical; Clinical Pathology; Complex; Complex Mixtures; Controlled Study; Coupling; Custom; Data; Detection; Development; Dextrans; Diagnosis; Diagnostic Neoplasm Staging; Dose; Electron Microscopy; Endotoxins; Environment; Equilibrium; Evaluation; Exclusion; Expert Opinion; Fatty Acids; Flow Cytometry; Foundations; Goals; Gold; Government; Hydrazine; Hydrazines; Hydrocarbons; Immunohistochemistry; Iron; Label; Laboratories; Ligands; Link; Magnetism; Magnetometries; Measurement; Measures; Methods; Micromanipulation; Molecular Analysis; Monitor; Normal Cell; Ohio; Particle Size; Pathologist; Pathology; Persons; Phase; Phase-Contrast Microscopy; Polymerase Chain Reaction; Population; Procedures; Process; Production; Property; Protein Chemistry; Protocols documentation; Quality Control; Reaction Time; Reagent; Recovery; Recurrence; Reproducibility; Research; Research Ethics Committees; Research Personnel; Resolution; Running; Sampling; Solutions; Sorting - Cell Movement; Source; Squid; Staining method; Stains; Sterility; Streptavidin; Surface; Suspension substance; Suspensions; System; Techniques; Testing; Tissue Procurements; Tissues; Trypan Blue; Universities; Validation; Variant; aqueous; base; cancer cell; carbonyl group; commercialization; density; design; dextran; disease diagnosis; functional group; head and neck cancer patient; innovation; instrument; interest; light scattering; magnetic cell separation; magnetic field; magnetic sorter; malignant breast neoplasm; manufacturing process; meetings; nanoparticle; nanopowder; neoplastic cell; novel; particle; peripheral blood; programs; prototype; quadrupole magnetic sorter; response; scale up; self assembly; tumor

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.

描述（由申请人提供）： 哥伦布纳米柯克斯（CNW）提出的研究导致定制磁性纳米颗粒试剂的商业化，以检测来自骨髓和外周血等各种来源的癌细胞。对于研究人员和临床医生来说，细胞分离变得越来越普遍，他们将稀有细胞与复杂细胞群体分离，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。复杂的混合物和细胞样品中的小变化需要新技术，这些技术具有高通量的精致灵敏度。因此，细胞分离技术的进步必须使研究人员能够找到感兴趣的低百分比细胞，并净化它们以准确评估生物学相关性。在第一阶段研究中，CNW证明了磁性纳米颗粒的生产，其大小均匀范围在50至70nm的范围内，其配体密度为2至5 ug/ml的Fe颗粒，并引起了免疫磁性标记的细胞的磁性迁移率，比商业上可用的磁性磁性大的含量更大。上述标准定义了在I期应用中获得的磁性颗粒的标准，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％的尖峰肿瘤样品，以及平均3.21 log10 log10耗竭的污染细胞的污染物耗尽，在流动性磁力剂中已知的Quadroupole磁性磁磁带（QMMS）。因此，第二阶段的研究要求对涉及癌细胞富集的QMS应用的磁试剂试剂盒的设计和商业化。 CNW将（1）扩展处理工作，以获得比当前过程技术增加50倍的增长； （2）建立程序和实践，以确保对纳米颗粒大小和磁性特性的一致控制； （3）为CNW细胞富集试剂盒开发用于商用磁流程QMS的CNW细胞富集套件； （4）使用乳腺癌患者以及头颈癌患者对方案进行测试。拟议的II期末端项目是针对QMS系统的定制磁试剂盒，该试剂盒将允许从各种血液源（例如骨髓和外周血）中富集癌细胞。颗粒的规格将提供超过70％的所需细胞恢复，以及4 log10污染细胞的排放。 QMS系统中使用CNW纳米颗粒的富集分数将提供质量纯化的产品，以进一步下游加工和随后的分子分析。目的是使CNW定制试剂成为当前病理库中黄金标准测试不可或缺的一部分。哥伦布纳米柯克斯是一家磁性纳米颗粒试剂公司，为磁细胞分离提供试剂。在此应用中，我们将开发磁试剂盒作为四极磁分落器（QMS）系统的商业试剂。过去，QMS系统中使用的商业磁性纳米颗粒在纯度和所需细胞的恢复方面提供了低于PAR的结果。在其第一阶段应用中证明的哥伦布纳米官能表明，为QMS设计的试剂能够达到平均污染细胞的平均3.21 log10耗竭，以及从对照样品中平均回收74.1％的肿瘤细胞。我们认为，CNW针对QMS系统的自定义试剂套件将为样本到样品提供更可预测的可再现结果。