Nanotrap technology for one step preservation and amplification of cancer biomark

Nanotrap 技术可一步保存和扩增癌症生物标志物

基本信息

批准号：
8433072
负责人：
Lance Allen Liotta
金额：
$ 38.04万
依托单位：
GEORGE MASON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-20 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8433072
关键词：
Advanced Development Affinity Albumins Biological Biological Assay Biological Markers Biological Preservation Blood Body Fluids Categories Chemicals Chemistry Clinical Collection Cystic Fibrosis Databases Detection Devices Diagnosis Diagnostic Neoplasm Staging Freezing Grant Harvest Human Hydrogels Immunoassay International Label Legal patent Licensing Liquid substance Magnetism Malignant Neoplasms Masks Mass Spectrum Analysis Measurement Measures Nanotechnology One-Step dentin bonding system Paralysed Penetration Peptides Performance Plasma Plasma Proteins Process Protease Inhibitor Proteins Publications Qualifying Research Running Sampling Science Serum Serum Proteins Specimen Sweat Sweating Technology Temperature Time Tube Tumor stage Urine Validation blind improved innovation multiple reaction monitoring nanoparticle new technology novel particle prevent scale up technology validation

项目摘要

DESCRIPTION (provided by applicant): We propose advanced development and validation of novel technology that will maximize the quality and utility of biologic fluid specimens, and permit the measurement of previously invisible low abundance biomarkers. The technology is transformative and paradigm-shifting because it immediately and economically solves fundamental roadblocks paralyzing the body fluid cancer biomarker field. In a single step, in minutes, our technology overcomes the severe problems of biomarker preservation, low abundance, and masking by unwanted proteins. The technology is novel porous, buoyant, core-shell hydrogel nanoparticles containing high affinity reactive chemical baits that harvests biomarkers in body fluids. Our nanoparticles can be pre loaded into Vacutainer(R) blood collection tubes, or other body fluid collection vessels. Upon contact with the sample, the suspended nanoparticles immediately affinity-sequester target biomarkers inside the particles, exclude albumin, fully protect the biomarkers from degradation (even at elevated temperatures), and massively concentrate the sequestered biomarkers into a small volume. The technology can dramatically (demonstrated up to 10,000 fold) improve the lower limits of detection and the precision of: a) mass spectrometry (MS) biomarker discovery, b) quantitation by multiple reaction monitoring (MRM), or c) quantification by any clinical grade immunoassay. All of the Aims and Milestones of the predicate IMAT NCI R21 CA137706 grant have been exceeded. We have discovered more than a dozen novel chemical baits with preferential high affinities (KD <10-11 M) for specific low abundance protein analytes. We discovered a novel shell chemistry that selectively prevented unwanted entry of all size albumin-derived peptides without hindering the penetration of non-albumin small proteins and peptides. Labile body fluid biomarkers, that would otherwise rapidly degrade, are completely preserved at the time of collection, thus obviating the need for costly freezing or proteinase inhibitors. Low abundant proteins previously invisible to discovery by MS, and previously not measureable by MRM, or clinical immunoassays, can now be quantified with high precision within the linear range of the assay. Our technology is completely innovative, as documented by 3 allowed (2 issued) patents that have been commercially licensed, and 14 publications. No existing technology can solve all of the aforementioned roadblocks, and attain a capture/elution efficiency near 100%. The technology has recently permitted the MS identification of new serum and plasma proteins not listed in the international HUPO database. Under the proposed Aims we will scale up the technology to conduct blind validation of its performance precision, accuracy, improved detection sensitivity, and preservation capacity, in two large (n = 400 sera and n = 74 plasma) well-controlled clinical sera/plasma sample sets (64 analytes). We will extend the technology to urine and sweat to open up these biofluids as a new category for biomarker research, using innovative approaches to solve fundamental problems of volume, perishability, and low protein concentration for these biofluids. PUBLIC HEALTH RELEVANCE: We propose advanced development and validation of novel nanotechnology that will maximize the quality and utility of biologic fluid specimens, and permit the measurement of previously invisible low abundance biomarkers. The nanotechnology is transformative and paradigm-shifting because it immediately and economically solves fundamental roadblocks paralyzing the body fluid cancer biomarker field, permitting the measurement of biomarkers emanating from small (< 2 mm) early stage cancers with high precision and accuracy.

描述（由申请人提供）：我们提出了新技术的高级开发和验证，该技术将最大化生物液标本的质量和实用性，并允许先前看不见的低丰度生物标志物的测量。该技术具有变革性和范式转移，因为它立即和经济地解决了基本障碍，使体液癌症生物标志物领域瘫痪。在一步之内，我们的技术克服了生物标志物保存，低丰度和掩盖不良蛋白质的严重问题。该技术是新型的多孔，浮力，核壳水凝胶纳米颗粒，其中包含高亲和力反应性化学诱饵，可收获体液中的生物标志物。我们的纳米颗粒可以预先装入空泡（R）血液收集管或其他体液收集容器中。与样品接触后，悬浮的纳米颗粒立即亲和力 - 靶标生物标志物在颗粒内，排除白蛋白，完全保护生物标志物免受降解（即使在升高的温度下），并将隔离的生物标记物大量浓缩到很小的体积中。该技术可以极大地（最多证明10,000倍）改善检测的下限和精度：a）质谱（MS）生物标志物发现，b）通过多个反应监测（MRM）或C）通过任何临床级别的免疫测定法进行定量。谓词IMAT NCI R21 CA137706赠款的所有目标和里程碑均已超过。我们发现了针对特定低丰度蛋白分析物的十几种具有优先高亲和力（KD <10-11 m）的新型化学诱饵。我们发现了一种新型的壳化学，有选择地防止了所有大小的白蛋白衍生肽的不必要进入，而不会阻碍非藻蛋白小蛋白和肽的穿透。否则会迅速降解的不稳定体液生物标志物在收集时被完全保存，从而消除了对昂贵的冷冻或蛋白酶抑制剂的需求。现在，MRM或MRM或临床免疫测定无法测量的先前不可见的低丰富蛋白质现在可以在测定的线性范围内用高精度来量化。正如已获得商业许可的3项允许（2项已发行的）专利和14个出版物所记录的那样，我们的技术是完全创新的。没有现有的技术可以解决上述所有障碍，并达到接近100％的捕获/洗脱效率。该技术最近允许MS鉴定国际HUPO数据库中未列出的新血清和血浆蛋白。在拟议的目标下，我们将扩大技术的规模，以对其性能精度，准确性，提高检测灵敏度和保存能力进行盲目验证，这是两个大的（n = 400种血清和n = 74个血浆）控制良好的临床血清/血浆样品集（64个分析）。我们将使用创新的方法将这些技术扩展到尿液和汗水，以开放这些生物流体，作为生物标志物研究的新类别，以解决这些生物流体的基本问题，易腐性和低蛋白质浓度的基本问题。公共卫生相关性：我们提出了新型纳米技术的高级开发和验证，这将最大程度地提高生物液标本的质量和实用性，并允许测量以前不可见的低丰度生物标志物。纳米技术具有变革性和范式转移，因为它立即和经济地解决了基本的障碍，使体液癌症生物标志物领域瘫痪，允许测量来自小的（<2 mm）早期阶段的生物标志物，具有高精度和准确性。