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

• 批准号: 8723137
• 负责人: Lance Allen Liotta
• 金额: $ 35.76万
• 依托单位: GEORGE MASON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-20 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8723137
• 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

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.

描述(由申请人提供)：我们建议进一步开发和验证新技术，以最大限度地提高生物体液样本的质量和效用，并允许 对以前不可见的低丰度生物标志物的测量。这项技术是变革性的和范式转换的，因为它立即和经济地解决了使体液癌生物标志物领域陷入瘫痪的根本障碍。在几分钟的时间里，我们的技术只需一个步骤，就可以克服生物标记物保存、丰度低和被不需要的蛋白质掩盖的严重问题。这项技术是一种新型的多孔、浮力、核壳水凝胶纳米颗粒，含有高亲和力的活性化学诱饵，可获取体液中的生物标记物。我们的纳米颗粒可以预装到Vacutainer(R)血液采集管或其他体液采集器中。一旦与样品接触，悬浮的纳米颗粒立即亲和隔离颗粒内的目标生物标记物，排除白蛋白，完全保护生物标记物不被降解(即使在高温下)，并将隔离的生物标记物大量浓缩到小体积中。该技术可以极大地提高检测下限和精确度：a)发现质谱学(MS)生物标记物，b)通过多反应监测(MRM)进行定量，或c)通过任何临床级别的免疫分析进行定量。已经超过了谓词IMAT NCI R21 CA137706授予的所有目标和里程碑。我们已经发现了十几种具有优先高亲和力(Kd&lt；10-11M)的新型化学诱饵，用于特定的低丰度蛋白质分析物。我们发现了一种新的壳化学，它可以选择性地阻止所有大小的白蛋白衍生多肽的不必要进入，而不会阻碍非白蛋白小蛋白和多肽的渗透。不稳定的体液生物标记物在收集时被完全保存下来，否则会迅速降解，因此不需要昂贵的冷冻或蛋白水解酶抑制剂。低丰度的蛋白质以前不能被MS发现，以前不能用MRM或临床免疫分析方法测量，现在可以在分析的线性范围内以高精度定量。我们的技术是完全创新的，已获得商业许可的3项(已颁发的)专利和14项出版物证明了这一点。没有一种现有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。这项技术最近允许对国际HuPo数据库中未列出的新血清和血浆蛋白质进行MS鉴定。在建议的目标下，我们将扩大该技术的规模，在两个控制良好的临床血清/血浆样本集(分析物)上对其性能精密度、准确性、改进的检测灵敏度和保存能力进行盲法验证。我们将把这项技术扩展到尿液和汗液，以开辟这些生物液作为生物标记物研究的新类别，使用创新的方法来解决这些生物液的体积、易腐烂和低蛋白质浓度等基本问题。