Nanotechnology based magnetic detection for rare cell assays

描述（由申请人提供）：MagneSensors的合作计划集中在一个新的基于纳米技术的平台上，该平台使用超灵敏的磁性传感器和磁性标签来执行强大的稀有细胞检测。该磁性检测平台使用高温超导量子干涉装置（SQUID）来检测用磁性纳米颗粒标记的抗体与细胞表面受体的结合。我们将应用该平台评估慢性淋巴细胞白血病（CLL）的微小残留病（MRD），以便更好地监测和指导治疗。一个主要的计划重点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。该仪器将用于证明能够快速检测CLL细胞的磁性测定。具体目标是：1）改进磁性纳米颗粒标记及其与抗体的缀合（dlOOnm尺寸，尺寸可变性<15%，缀合后聚集减少10倍以检测Ab，血清中的低聚集）2）构建用于稀有细胞检测的新仪器（灵敏度提高10倍，20倍更高的通量）3）用新仪器和试剂展示在简单CLL模型系统中对稀有细胞的超灵敏检测，与流式细胞术比较（在107个细胞的背景中检测50个细胞<45分钟）4）证明快速，灵敏磁细胞法检测慢性淋巴细胞白血病微小残留病（在<45分钟内检测107个白细胞中少至50个CLL细胞）磁性平台的高灵敏度使得能够检测稀有细胞（1/104-106）或表达少量受体的细胞。混合和测量格式显著缩短了测定制备时间并简化了自动化（无洗涤步骤）。这种在很少或没有样品制备的情况下工作的能力促进了对临床样品的快速、高通量测试。跨学科的努力联合了Moores UCSD癌症中心和纳米肿瘤卓越中心的癌症纳米技术，UCSD的白血病应用，Immunicon公司的定制磁性纳米颗粒标签和罕见细胞检测专业知识，SoluLink公司。用于将抗体连接到磁性纳米颗粒的生物缀合化学，以及MagneSensors，Inc.慢性淋巴细胞白血病（CLL）是北美和欧洲最常见的白血病，占所有白血病的30%以上。CLL仍然无法用目前可用的治疗方法治愈，患者的预后差异很大。现有的技术，如流式细胞术和聚合酶链反应（PCR），可用于评估微小残留病变，以优化治疗，是不足以常规临床使用。能够快速，超灵敏，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，并通过使医生能够比目前更快地调整治疗方法来改善患者预后。

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.