A non-invasive nanotechnology-based skin patch for multiplexed diagnostics

基于纳米技术的非侵入性皮肤贴片，用于多重诊断

• 批准号: 8092239
• 负责人: Lance Allen Liotta
• 金额: $ 16.37万
• 依托单位: GEORGE MASON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8092239
• 关键词: Adhesives; Affinity; Albumins; Animal Model; Artificial nanoparticles; Binding; Biological; Biological Assay; Biological Markers; Biological Preservation; Calcitonin; Cavia; Chemicals; Chemistry; Clinical; Collection; Cystic Fibrosis; Detection; Devices; Diagnostic; Diagnostic tests; Disease; Drug Delivery Systems; Dyes; Electrolytes; Estradiol; Evaluation; Exclusion; FDA approved; Feasibility Studies; Female; Foundations; Grant; Harvest; Hour; Human; Human Volunteers; Hydrogels; IL8 gene; Immunoassay; Incubated; Institutional Review Boards; Interleukin-6; Interleukins; Iontophoresis; Liquid substance; Mails; Mass Spectrum Analysis; Measurement; Measures; Membrane; Modeling; Molecular Weight; Nanotechnology; New Zealand; One-Step dentin bonding system; Oryctolagus cuniculus; Particle Size; Patch Tests; Patients; Peptide Hydrolases; Peptides; Physiological; Process; Progesterone; Proteins; Running; Sampling; Screening procedure; Serum; Skin; Solutions; Source; Surface; Sweat; Sweating; TNF gene; Technology; Temperature; Testing; Testosterone; Textiles; Thick; Time; Toxic effect; Transudate; base; chemical release; disease diagnosis; eccrine; healthy volunteer; interest; male; nanoparticle; nervous system disorder; new technology; novel; particle; poly-N-isopropylacrylamide; prevent; protein metabolite; skin irritation; skin patch; uptake; volunteer

DESCRIPTION (provided by applicant): Sweat is an unexploited biological fluid that can provide a wealth of diagnostic information. We propose a novel technology to be used at the bedside or in the field: a diagnostic skin patch which harvests, concentrates, and stabilizes a panel of biomarkers derived from skin transudate or sweat. While drug delivery patches are routinely used, the technology proposed here has exactly the opposite function: the harvesting of diagnostic markers. Using mass spectrometry we have identified 228 proteins and peptides that were not previously known to exist in human sweat. In order to exploit this new class of analytes we propose to create novel affinity bait nanoparticles, bound within an adhesive skin patch. The proposed technology is transformative because it overcomes all the major physiological barriers that have prevented the use of this biologic fluid for diagnostic testing. Sweat disease biomarkers a) are subject to rapid degradation due to proteases present in sweat and normal skin bacterial flora, and b) exist in extremely low abundance, far below the detection sensitivity of standard analysis platforms. Harvesting hydrogel nanoparticles are engineered with chemical high affinity baits so that they sequester the low abundance target analytes, and protect them from degradation indefinitely. We propose to integrate the nanoparticles into the fabric of an adhesive skin patch. Once applied to the skin, the nanoparticles in the patch harvest minute by minute, and protect from degradation, all candidate analytes in the sweat underneath the patch. The core shell bait nanoparticles are a completely novel technology that can amplify the sensitivity of biomarker detection by 100 fold. No other technology exists that has a similar yield, concentration ability, and stabilization function. Once the collection is complete, the patch can simply be mailed to the diagnostic lab at room temperature. Upon receipt, the nanoparticle-captured analytes of interest can be eluted from the patch for routine measurement using any platform. Our feasibility studies demonstrate virtually 100 percent capture and 100 percent elution yield of low abundance interleukins in model sweat solutions. We will engineer the nanoparticles, and construct test patch devices. The test patches will be evaluated in animal models to verify lack of skin irritation. We will collect sweat from healthy volunteers under IRB approval using an FDA approved iontophoresis sampling device used for electrolyte measurement. We will apply the collected sweat to the novel nanoparticle patch ex vivo at the point of collection. Mass spectrometry will be used to discover novel sweat biomarkers that have been concentrated and preserved in the patch. Low abundance labile sweat biomarkers harvested from the nanoparticles will be measured by clinical immunoassays to verify sensitivity and precision. The derived list of eccrine sweat proteins will be an important deliverable as a foundation for the general field of sweat biomarker testing. The technology is especially suited to the evaluation of neurological disorders as it is non invasive and would be fully acceptable as a routine screening procedure. PUBLIC HEALTH RELEVANCE: The low abundance and low molecular weight proteins and metabolites present in human sweat provide great promise as a source of new biomarkers for neurological disease diagnosis. The nanotechnology proposed in this grant will greatly reduce the preanalytical variability associated with collection and storage of biological fluids for clinical analysis and at the same time allow for detection of low abundance and labile analytes otherwise not possible.

描述(申请人提供)：汗液是一种未开发的生物流体，可以提供丰富的诊断信息。我们提出了一种用于床边或现场的新技术：一种诊断性皮肤贴片，它收集、浓缩和稳定一组来自皮肤渗出液或汗液的生物标记物。虽然药物输送贴片是常规使用的，但这里提出的技术具有完全相反的功能：获取诊断标记。利用质谱学，我们已经鉴定出228种蛋白质和多肽，这些蛋白质和多肽以前不存在于人类汗液中。为了开发这类新的分析物，我们建议创造新的亲和诱饵纳米颗粒，结合在粘性皮肤贴片中。这项拟议的技术具有变革性，因为它克服了阻碍使用这种生物液体进行诊断测试的所有主要生理障碍。汗病生物标志物a)由于汗液和正常皮肤细菌菌群中存在的蛋白酶而迅速降解，b)以极低的丰度存在，远远低于标准分析平台的检测灵敏度。收集水凝胶纳米颗粒是用化学高亲和力诱饵设计的，因此它们隔离了低丰度的目标分析物，并无限期地保护它们免受降解。我们建议将纳米颗粒集成到粘性皮肤贴片的织物中。一旦涂在皮肤上，贴片中的纳米颗粒就会一分钟一分钟地收集起来，防止贴片下面汗液中的所有候选分析物降解。核壳诱饵纳米颗粒是一项全新的技术，可以将生物标志物检测的灵敏度放大100倍。没有其他技术具有类似的产量、浓缩能力和稳定功能。一旦收集完成，补丁只需在室温下邮寄到诊断实验室即可。一旦收到，感兴趣的纳米颗粒捕获的分析物可以从贴片中洗脱出来，用于使用任何平台进行常规测量。我们的可行性研究表明，在模型汗液中，几乎100%捕获和100%洗脱低丰度白细胞介素得率。我们将设计纳米粒子，并建造测试贴片设备。测试贴片将在动物模型中进行评估，以验证是否没有皮肤刺激性。我们将使用FDA批准的用于电解质测量的离子导入采样设备，从IRB批准的健康志愿者身上收集汗液。我们将收集的汗液应用于新型纳米颗粒贴片的体外采集点。质谱仪将用于发现新的汗液生物标记物，这些标记物已被浓缩并保存在贴片中。从纳米颗粒中获取的低丰度不稳定汗液生物标记物将通过临床免疫分析进行测量，以验证敏感性和精密度。作为汗液生物标记物测试一般领域的基础，汗液分泌蛋白质的衍生清单将是一项重要的成果。这项技术特别适用于神经疾病的评估，因为它是非侵入性的，完全可以作为常规筛查程序接受。 与公共健康相关：人类汗液中存在的低丰度和低分子质量的蛋白质和代谢物为神经疾病诊断的新生物标志物提供了巨大的希望。这项赠款中提议的纳米技术将大大降低与收集和储存用于临床分析的生物流体相关的分析前可变性，同时允许检测低丰度和不稳定的分析物，否则是不可能的。