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在新的超灵敏夹心免疫测定中使用磁性传感器和磁性纳米颗粒标记。重点是使用非常理想的混合和测量格式测量血液中非常低的抗原水平。这种组合极难实现，目前在竞争性生物测定平台中不可用。磁性检测提供了独特的优势，例如结合的定量测量，而不需要通常分离未结合的标记，以及血液和血清中固有的磁性背景的缺乏。该磁性检测平台使用高温超导量子干涉装置（SQUID）来检测与磁性纳米颗粒标记的抗体的抗原的特异性结合。重要的是，夹心免疫测定的超高灵敏度将利用我们新的下一代磁性检测仪器，该仪器将在单独的NIH计划下很快上线。这款新型台式仪器使用小型低温制冷机，信噪比比我们现有的仪器高出沿着50- 200倍，通量也高得多。高灵敏度提供了早期检测和测量小体积有价值的试剂或稀缺样品的优势。混合和测量格式显著减少了测定制备时间并简化了自动化（无洗涤步骤），从而实现快速、高通量测定。在血液和血清中工作的能力为更广泛的重要免疫测定带来了这些益处。I期的目的是证明在白介素-6（IL-6）模型系统上的超高灵敏度磁性夹心免疫测定，具有以下目标：1）全血，2）<0.4pg/ml，3）未结合的磁性纳米颗粒标记物不分离，4）包括制备和测量的总测定时间<60分钟。第一阶段的重点是使用夹心磁性免疫测定格式检测血液中的抗原。0.4 pg/ml的检测目标是血液中内源性IL-6水平的下限。这将在第一阶段使用我们开发的一种新技术来实现，该技术使用二氧化硅微珠和密度梯度介质在磁检测之前从血细胞中去除捕获的抗原。请注意，第一阶段的努力并没有开发出真正的混合和测量血液中的免疫测定，因为在第一阶段中，血细胞是从捕获的抗原中分离出来的。这个更雄心勃勃的目标，目前无法实现超高灵敏度与其他检测平台，是留给第二阶段。