Ultrabright Fluorescent Nanoconstruct Enabling Widely Accessible, High Performance Multiplexed Protein Assays

超亮荧光纳米结构实现广泛可及的高性能多重蛋白质检测

• 批准号: 10080340
• 负责人: Scott L Crick
• 金额: $ 77.63万
• 依托单位: AURAGENT BIOSCIENCE, LLC
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-17 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10080340
• 关键词: Adoption; Aging; Antibodies; Biochemical; Biological; Biological Assay; Biological Markers; Biomedical Research; Capital; Clinical Research; Complex; Computer software; Data Analyses; Detection; Disease; Elements; Ensure; Enzyme-Linked Immunosorbent Assay; Equipment; Fluorescence; Freeze Drying; Glass; Goals; Health; Image; Immunoassay; Label; Laboratory Research; Lasers; Life; Light; Liquid substance; Malignant Neoplasms; Measurement; Measures; Methods; Microarray Analysis; Neurodegenerative Disorders; Performance; Price; Procedures; Protein Analysis; Proteins; Proteome; Proteomics; Protocols documentation; Pyroxylin; Reader; Reading; Reporter; Research; Research Personnel; Role; Sampling; Sapphire; Signal Transduction; Source; Streptavidin; System; Testing; Therapeutic Intervention; Tissues; Translations; Western Blotting; Work; base; clinical diagnostics; complex biological systems; cost; cytokine; design; fluorophore; improved; instrumentation; plasmonics; programs; prototype; research and development; response; tool; user-friendly

Abstract Detection and quantification of multiple proteins in biological fluids and tissues is of fundamental importance in biomedical research and clinical diagnostics because it is impossible to understand complex, non-linear, biochemical systems without being able to accurately interrogate the components. The need to interrogate multiple proteins simultaneously is ubiquitous across all domains of biomedical research, and it is a major barrier to fully understanding health, ageing, disease, and response to therapeutic interventions. Though the need may be ubiquitous, there is not a widely accessible solution for researchers to make multiplexed protein measurement with high sensitivity and a large dynamic range. Antibody microarrays have a straightforward, ELISA-like workflow and can be used to measure up to thousands of proteins simultaneously. Unfortunately, they have rather poor sensitivity, and require a specialized and expensive (>$75k) reader to be used. These are significant barriers to adoption for the majority of biomedical researchers. We have developed an ultrabright fluorescent nanoconstruct we call the Plasmonic Fluor, which is >5,000X brighter than the standard fluorescent reporter used in microarrays today. Simply substituting the Plasmonic Fluor for the existing reporter significantly increases the sensitivity of antibody microarrays without requiring any change to the workflow. Importantly, the Plasmonic Fluor is so bright that it allows fluorescent microarrays to be read using more widely available and versatile readers such as a fluorescent Western blot reader. Additionally, it relaxes the requirements on the reader instrumentation significantly, which has enabled us to create an inexpensive reader that any research lab can afford. To allow even easier adoption of the Plasmonic Fluor enhanced microarrays, we have also created software for microarray data analysis. In this project, we aim to: increase the manufacturing scale of our Plasmonic Fluor to an early commercial scale; more extensively validate enhancement of a variety of popular antibody microarrays; finalize the design of our inexpensive reader; and more fully develop our user- friendly microarray analysis software. We believe the Plasmonic Fluor will become the standard fluorescent reporter molecule for all microarrays, and Plasmonic Fluor-enhanced microarrays will become a widely used and powerful tool for elucidating the role of protein networks in health and disease.

抽象的 生物体液和组织中多种蛋白质的检测和定量至关重要 在生物医学研究和临床诊断中的重要性，因为它是不可能理解的 复杂的、非线性的生化系统，而无法准确地询问 成分。同时询问多种蛋白质的需求在所有领域中都普遍存在。 生物医学研究领域，它是充分了解健康、衰老、 疾病以及对治疗干预的反应。尽管这种需求可能无处不在， 对于研究人员来说，这并不是一种广泛可用的制备多重蛋白质的解决方案 测量灵敏度高，动态范围大。 抗体微阵列具有类似 ELISA 的简单工作流程，可用于测量 同时处理多达数千个蛋白质。不幸的是，它们的灵敏度相当差，并且 需要使用专门且昂贵的（> 75,000 美元）阅读器。这些都是重大障碍 为大多数生物医学研究人员所采用。我们开发了一种超亮荧光灯 我们称之为等离子荧光体的纳米结构，其亮度比标准值高 5,000 倍以上 如今用于微阵列的荧光报告基因。只需用等离激元荧光代替 现有的报告器显着提高了抗体微阵列的灵敏度，而不需要 对工作流程的任何更改。重要的是，等离子体荧光非常明亮，可以发出荧光 使用更广泛使用和多功能的阅读器（例如荧光阅读器）来阅读微阵列 蛋白质印迹阅读器。此外，它还放宽了对读卡器仪器的要求 显着，这使我们能够创造出任何研究实验室都可以使用的廉价阅读器 买得起。为了更容易地采用等离子荧光增强微阵列，我们有 还创建了微阵列数据分析软件。 在这个项目中，我们的目标是： 将等离子荧光剂的制造规模扩大到早期 商业规模；更广泛地验证多种流行抗体的增强作用 微阵列；完成我们廉价阅读器的设计；并更充分地开发我们的用户 友好的微阵列分析软件。 我们相信等离子荧光将成为所有标准的荧光报告分子 微阵列和等离子体荧光增强微阵列将成为一种广泛使用且强大的技术 阐明蛋白质网络在健康和疾病中的作用的工具。