Ultra high sensitivity mix and measure immunoassays in blood

超高灵敏度混合并测量血液中的免疫测定

• 批准号: 7482524
• 负责人: Mark S. DiIorio
• 金额: $ 16.11万
• 依托单位: MAGNESENSORS, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7482524
• 关键词: Aliquot; Antibodies; Antigens; Area; Automation; Basic Science; Binding; Biological Assay; Biological Models; Biotechnology; Blood; Blood Cells; Blood Tests; Blood specimen; Buffers; Centrifugation; Clinical; Complex; Contracts; Detection; Development; Devices; Diagnostic; Early Diagnosis; Figs - dietary; Fingers; Foundations; Generations; Goals; Head; High temperature of physical object; Immunoassay; In Vitro; Incubated; Inflammatory Response; Institutes; Interleukin-6; Label; Left; Legal patent; Liquid substance; Magnetism; Measurement; Measures; Melanocytic nevus; Microspheres; Mole the mammal; Monitor; Monoclonal Antibodies; Nature; Noise; Personal Satisfaction; Phase; Preparation; Principal Investigator; Publications; Range; Reagent; Research; Sampling; Sensitivity and Specificity; Serum; Signal Transduction; Silicon Dioxide; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Specificity; System; Techniques; Technology; Testing; Time; United States National Aeronautics and Space Administration; United States National Institutes of Health; Whole Blood; Work; Writing; antibody conjugate; authority; base; commercial application; concept; cost; cytokine; density; design; drug discovery; experience; fiberglass; high throughput screening; improved; in vivo; innovation; instrument; instrumentation; nanoparticle; next generation; novel; programs; prototype; research and development; sensor; size; superconducting quantum interference device; technology development

DESCRIPTION (provided by applicant): MagneSensors' Phase I SBIR uses magnetic sensors and magnetic nanoparticle labels in new ultra-sensitive sandwich immunoassays. The focus is on measuring very low levels of antigens in blood using the highly desirable mix and measure format. This combination is extremely difficult to achieve and is currently not available in competing bioassay platforms. Magnetic detection offers unique advantages, such as the quantitative measurement of binding without requiring the usual separation of unbound labels, as well as the inherent lack of magnetic background in blood and serum. This magnetic detection platform uses high temperature superconducting quantum interference devices (SQUIDs) to detect the specific binding to antigens of antibodies labeled with magnetic nanoparticles. Importantly, the ultra-high levels of sensitivity for sandwich immunoassays will take advantage of our new, next-generation magnetic detection instrument that will be brought on line shortly under a separate NIH program. This new bench top instrument uses a small cryorefrigerator and offers 50-200X better signal-to-noise than our existing instrument along with much higher throughput. High sensitivity offers advantages of early detection and measurements on small volumes of valuable reagents or scarce samples. The mix and measure format significantly reduces assay preparation time and simplifies automation (no wash steps), thereby enabling rapid, high throughput assays. The ability to work in blood and serum brings these benefits to a much broader range of important immunoassays. The Phase I aim is to demonstrate ultra high sensitivity magnetic sandwich immunoassays on an interleukin-6 (IL-6) model system with the following goals: 1) whole blood, 2) <0.4 pg/ml, 3) no separation of unbound magnetic nanoparticle labels, 4) <60 minutes total assay time including preparation and measurement. Phase I focuses on detection of an antigen in blood using a sandwich magnetic immunoassay format. The detection goal of 0.4 pg/ml is at the lower end of endogenous levels of IL-6 in blood. It will be achieved in Phase I using a novel technique we have developed that uses silica microbeads and a density gradient media to remove the captured antigen from blood cells prior to magnetic detection. Note that the Phase I effort does not develop a true mix and measure immunoassay in blood, as blood cells are separated out from captured antigen in Phase I. This more ambitious goal, currently unachievable at ultra-high sensitivity with other detection platforms, is left for Phase II.

描述（由申请人提供）：Magnesensors的I期SBIR在新的超敏感三明治免疫测定中使用磁性传感器和磁性纳米颗粒标签。重点是使用高度理想的混合和测量格式测量血液中非常低的抗原。这种组合非常困难，目前在竞争的生物测定平台中不可用。磁检测具有独特的优势，例如结合的定量测量，而无需通常的未结合标签，以及血液和血清中固有的磁背景固有的缺乏。该磁性检测平台使用高温超导量子干扰装置（鱿鱼）来检测与磁性纳米颗粒标记的抗体的特异性结合。重要的是，三明治免疫测定的超高敏感性水平将利用我们的新的下一代磁性检测仪器，该工具将在不久的将单独的NIH程序下进行线条。这种新的台式仪器使用小型冷冻冰箱，比我们现有的仪器和更高的吞吐量提供50-200倍的信噪比。高灵敏度提供了早期检测和测量的优势，并在少量有价值的试剂或稀缺样本上进行了测量。混合和测量格式大大减少了测定时间的准备时间并简化自动化（无需洗涤步骤），从而实现快速，高吞吐量测定。在血液和血清中工作的能力为这些好处带来了更广泛的重要免疫测定。第一阶段的目的是在白介素-6（IL-6）模型系统上证明具有以下目标的超高灵敏度磁性夹心免疫测定：1）全血，2）<0.4 pg/ml，3）无抗磁性纳米颗粒标签的分离，4）<60分钟<60分钟<60分钟<60分钟<60分钟<60分钟的总测定时间。第一阶段的重点是使用三明治磁性免疫测定格式检测血液中的抗原。 0.4 pg/mL的检测目标位于血液中IL-6内源水平的下端。它将在第一阶段使用我们开发的新技术来实现，该技术在磁性检测之前使用二氧化硅微粒和密度梯度培养基从血细胞中去除捕获的抗原。请注意，第一阶段的努力不会产生真正的混合物，并测量血液中的免疫测定法，因为在第一阶段的血细胞与捕获的抗原分开。这个更雄心勃勃的目标目前在具有其他检测平台的超高敏感性下无法实现。