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

基本信息

批准号：
7238062
负责人：
Arfaan Rampersaud
金额：
$ 16.7万
依托单位：
COLUMBUS NANOWORKS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-09-01 至 2009-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7238062
关键词：
Antibodies Binding Biological Biotin Blood Blood Cells Bone Marrow Buffers Cancer Diagnostics Cancer Patient Carbon Cell Line Cell Separation Cell Survival Cell physiology Cells Chemicals Clinic Clinical Complex Complex Mixtures Controlled Study Coupling Custom Data Detection Development Dextrans Diagnosis Diagnostic Neoplasm Staging Disease Electron Microscopy Endotoxins Ensure Environment Equilibrium Evaluation Exclusion Fatty Acids Flow Cytometry Foundations Goals Gold Government Hydrazine Hydrazines Hydrocarbons Instruction Iron Label Laboratories Laboratory Technicians Ligands Link Magnetism Magnetometries Measurement Measures Methods Molecular Analysis Monitor National Cancer Institute Ohio Oleate Particle Size Pathology Persons Phase Polymerase Chain Reaction Population Procedures Process Production Property Protein Chemistry Protocols documentation Purpose Quality Control Range Rate Reaction Time Reagent Recovery Recurrence Research Research Ethics Committees Research Personnel Resolution Sampling Silicon Dioxide Solutions Sorting - Cell Movement Source Specificity Squid Standards of Weights and Measures Sterility Streptavidin Surface Suspension substance Suspensions System Techniques Testing Tissues Training Trypan Blue Universities Variant Week aqueous base cancer cell cancer diagnosis carbonyl group clinical application commercialization density design desire dextran functional group head and neck cancer patient human tissue innovation instrument interest light scattering magnetic cell separation magnetic field malignant breast neoplasm manufacturing process nanoparticle nanopowder neoplastic cell novel particle peripheral blood programs prototype scale up self assembly size tissue processing tumor

项目摘要

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 NanoWorks (CNW) 提出了一项研究，旨在将定制磁性纳米颗粒试剂商业化，用于检测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，他们从复杂的细胞群中分离出稀有细胞，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技术，以提供高通量和精致的灵敏度；因此，细胞分离技术的进步必须使研究人员能够找到低百分比的感兴趣细胞，并纯化它们以准确评估生物学相关性。在第一阶段研究中，CNW 展示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，并引发免疫磁性标记细胞的磁泳迁移率一个数量级大于市售磁性试剂。上述第一阶段应用中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，加标肿瘤样品的平均回收率为 74.1%，并且流中的污染细胞平均去除 3.21 log10 -直通式磁力分选机，称为四极磁力分选机 (QMS)。因此，第二阶段研究需要设计和商业化专门针对涉及癌细胞富集的 QMS 应用的磁性试剂盒。 CNW 将 (1) 扩大加工力度，使其比当前加工技术提高 50 倍； (2) 建立确保纳米颗粒尺寸和磁性能一致控制的程序和实践； (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； (4)使用乳腺癌患者和头颈癌患者对方案进行β测试。拟议的第二阶段最终项目是专门针对 QMS 系统的定制磁性试剂盒，可以从骨髓和外周血等各种血液来源富集癌细胞。颗粒的规格将提供超过 70% 的所需细胞回收率，以及 4 log10 污染细胞的去除率。在 QMS 系统中使用 CNW 纳米粒子所得的富集级分将为进一步的下游处理和后续分子分析提供优质纯化的产品。 CNW 定制试剂的目标是成为当前病理学体系中金标准测试不可或缺的一部分。 Columbus NanoWorks 是一家磁性纳米颗粒试剂公司，提供用于磁性细胞分离的试剂。在此应用中，我们将开发磁性试剂套件作为四极磁分选机 (QMS) 系统的商业试剂。过去，QMS 系统中使用的商业磁性纳米粒子在所需细胞的纯度和回收率方面提供了低于标准的结果。 Columbus NanoWorks 在其第一阶段应用中证明，为 QMS 定制设计的试剂能够实现平均 3.21 log10 的污染细胞去除率，以及从对照样品中平均回收 74.1% 的肿瘤细胞。我们相信 CNW 的 QMS 系统定制试剂盒将为样品提供更可预测、可重复的结果。