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: 7583976
• 负责人: Arfaan Rampersaud
• 金额: $ 30.7万
• 依托单位: COLUMBUS NANOWORKS, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2011-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7583976
• 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)： 哥伦布纳米工厂(CNW)建议进行研究，使定制的磁性纳米颗粒试剂商业化，以检测来自各种来源的癌细胞，如骨髓和外周血液。对于从复杂细胞群中分离稀有细胞以了解细胞在不同环境中的功能或诊断疾病发生、复发或进展的研究人员和临床医生来说，细胞分离正变得越来越常见。复杂的混合物和对细胞样本中微小变异的搜索需要新的技术来提供精致的灵敏度和高通量；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，并对它们进行纯化，以便准确评估生物学相关性。在第一阶段的研究中，CNW证明了磁性纳米颗粒的制造，这种纳米颗粒的尺寸在50到70纳米范围内是均匀的，具有2到5微克/毫升的铁颗粒的配体密度，并引起免疫磁性标记细胞的磁电泳性，比商业上可用的磁性试剂大一个数量级。上述定义在第一阶段应用中获得的磁性颗粒的标准允许在107个总细胞中浓缩1个肿瘤细胞，平均回收率为74.1%的添加的肿瘤样本，以及在称为四极磁分选器(QMS)的流通式磁性分选机中污染细胞的平均3.21log10耗竭。因此，第二阶段的研究需要设计专门用于涉及癌细胞浓缩的QMS应用的磁性试剂盒并将其商业化。CNW公司将(1)扩大加工努力，以使目前的加工技术增加50倍；(2)建立程序和实践，确保对纳米颗粒尺寸和磁性的一致控制；(3)开发用于商业磁流分选机QMS的CNW细胞浓缩试剂盒；以及(4)对乳腺癌患者和头颈癌患者进行协议的Beta测试。拟议的第二阶段成品是专用于QMS系统的定制磁性试剂盒，将允许从各种血液来源(如骨髓和外周血液)浓缩癌细胞。颗粒的规格将提供超过70%的所需细胞回收率，以及4log10污染细胞的耗尽率。在QMS系统中使用CNW纳米颗粒得到的富集组分将为后续加工和后续的分子分析提供高质量的纯化产品。我们的目标是让CNW定制试剂成为当前病理剧目中金标准测试的组成部分。Columbus NanWorks是一家提供磁性细胞分离试剂的磁性纳米颗粒试剂公司。在这项应用中，我们将开发磁性试剂盒，作为四极磁分选器(QMS)系统的商业试剂。过去，在QMS系统中使用的商用磁性纳米颗粒在所需细胞的纯度和回收率方面提供了低于平均水平的结果。Columbus NanWorks在他们的第一阶段应用中证明，为QMS定制的试剂能够实现污染细胞的平均3.21log10消耗，以及从对照样本中平均回收74.1%的肿瘤细胞。我们相信，CNW用于QMS系统的定制试剂盒将在不同样本之间提供更可预测、更具重复性的结果。