Nanotechnology based magnetic detection for rare cell assays

描述（由申请人提供）：MagneSensors的合作项目以一种新型纳米技术为基础的平台为中心，该平台使用超灵敏的磁传感器和磁标签来执行强大的稀有细胞分析。该磁检测平台利用高温超导量子干涉器件（squid）检测磁性纳米颗粒标记的抗体与细胞表面受体的结合。我们将把该平台应用于慢性淋巴细胞白血病（CLL）的微小残留病（MRD）评估，以便更好地监测和指导治疗。一个主要项目的推力是开发改进的磁传感器和磁性纳米粒子标签，并将它们纳入下一代仪器。该仪器将用于演示能够快速检测CLL细胞的磁分析。具体目标是：1)改进磁性纳米颗粒标记及其与抗体的偶联（d100 nm尺寸，尺寸变异性<15%，偶联后抗体聚集减少10倍，血清聚集低）2)建立新的罕见细胞检测仪器（灵敏度提高10倍，吞吐量提高20倍）3)用新的仪器和试剂在简单CLL模型系统中实现罕见细胞的超灵敏检测。与流式细胞术相比（在107个细胞的背景中检测50个细胞<45分钟）4)证明快速、灵敏的磁细胞法用于临床样本中慢性淋巴细胞白血病的最小残留疾病评估（在<45分钟内检测107个白细胞中少至50个CLL细胞）磁平台的高灵敏度可以检测到罕见细胞（104-106个中有1个）或表达少量受体的细胞。混合和测量格式显著减少了分析准备时间，简化了自动化（无洗涤步骤）。这种工作能力很少或没有样品制备，有利于快速，高通量的临床样品检测。跨学科的努力联合了Moores UCSD癌症中心和癌症纳米技术纳米肿瘤卓越中心，UCSD用于白血病应用，Immunicon公司用于定制磁性纳米颗粒标签和稀有细胞检测专业知识，SoluLink公司用于将抗体附着在磁性纳米颗粒上的生物偶联化学，以及MagneSensors公司用于磁性传感器，仪器和磁性分析专业知识。慢性淋巴细胞白血病（CLL）是北美和欧洲最常见的白血病形式，占所有白血病的30%以上。CLL仍然无法治愈，目前可用的治疗方法和患者的预后差异很大。现有的用于评估最小残留疾病以优化治疗的技术，如流式细胞术和聚合酶链反应（PCR），不足以用于常规临床应用。一种能够快速、超灵敏地检测罕见白血病细胞的磁性平台可以评估CLL中最小的可检测疾病，并且通过使医生能够比目前更快地调整治疗方法，具有改善患者预后的临床效用。

DESCRIPTION (provided by applicant): MagneSensors' collaborative program is centered on a novel nanotechnology based platform that uses ultra-sensitive magnetic sensors and magnetic labels to perform powerful rare cell assays. This magnetic detection platform uses high temperature superconducting quantum interference devices (SQUIDs) to detect the binding to cell surface receptors of antibodies labeled with magnetic nanoparticles. We will apply the platform to the assessment of minimal residual disease (MRD) in chronic lymphocytic leukemia (CLL) in order to better monitor and guide therapies. A major program thrust is to develop improved magnetic sensors and magnetic nanoparticle labels and incorporate them into next generation instrumentation. This instrumentation will be used to demonstrate magnetic assays capable of rapidly detecting CLL cells. The specific aims are: 1) Improve magnetic nanoparticle labels and their conjugation to antibodies (d100 nm size with <15% size variability, 10X reduced aggregation after conjugation to detect Ab, low aggregation in serum) 2) Build new instrument for rare cell detection (10X improved sensitivity, 20X higher throughput) 3) Demonstrate ultra-sensitive detection of rare cells in simple CLL model system with new instrument and reagents, compare to flow cytometry (detect 50 cells in background of 107 cells <45 minutes) 4) Demonstrate rapid, sensitive magnetic cell assay for minimal residual disease assessment of chronic lymphocytic leukemia in clinical samples (detect as few as 50 CLL cells in 107 leukocytes in <45 min) The high sensitivity of magnetic platform enables the detection of rare cells (1 in 104-106), or cells expressing a low number of receptors. The mix and measure format significantly reduces assay preparation time and simplifies automation (no wash steps). This ability to work with little or no sample preparation facilitates rapid, high throughput testing on clinical samples. The interdisciplinary effort unites Moores UCSD Cancer Center and Nano Tumor Center of Excellence for Cancer Nanotechnology, UCSD for the leukemia application, Immunicon Corporation for the custom magnetic nanoparticle labels and rare cell detection expertise, SoluLink, Inc. for the bioconjugation chemistry for attaching antibodies to magnetic nanoparticles, and MagneSensors, Inc. for magnetic sensors, instrumentation, and magnetic assay expertise.Project Narrative Chronic lymphocytic leukemia (CLL) is the most common form of leukemia in North America and Europe and accounts for over 30% of all leukemias. CLL remains incurable with the currently available treatments and the prognosis of patients varies substantially. Existing techniques available to assess minimal residual disease to optimize treatment, such as flow cytometry and polymerase chain reaction (PCR), are inadequate for routine clinical use. A magnetic platform capable of rapid, ultra-sensitive, assays to detect rare leukemia cells could assess minimal detectable disease in CLL and have clinical utility for improving patient outcomes by enabling physicians to adjust therapies sooner than is currently possible.