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: 10194377
• 负责人: Scott L Crick
• 金额: $ 80.77万
• 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/10194377
• 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; 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; detection method; detection sensitivity; fluorophore; improved; instrumentation; multiplex assay; 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 在生物流体和组织中的多种蛋白质的检测和定量是基本的 在生物医学研究和临床诊断中的重要性，因为无法理解 复杂的，非线性的生化系统，无法准确审问 成分。在所有人的情况下，需要同时询问多种蛋白质的需求是无处不在的 生物医学研究领域，这是完全了解健康，衰老， 疾病以及对治疗干预措施的反应。尽管需求可能无处不在，但 对于研究人员制造多重蛋白的解决方案不是广泛的解决方案 具有高灵敏度和较大动态范围的测量。 抗体微阵列具有直接的，类似ELISA的工作流程，可用于测量 同时，多达数千种蛋白质。不幸的是，它们的敏感性相当差，并且 需要使用专业且昂贵（> $ 75K）的读者。这些是重大的障碍 大多数生物医学研究人员的收养。我们已经开发了一个超操荧光 纳米构造我们称之为等离荧光，比标准更明亮5,000倍 今天在微阵列中使用的荧光记者。只需取代等离子体荧光代替 现有的记者显着提高了抗体微阵列的灵敏度，而无需 工作流程的任何更改。重要的是，等离子体荧光是如此明亮，以至于荧光灯 使用更广泛的读取器（例如荧光）来读取的微阵列 Western印迹阅读器。此外，它放松了读取器仪器的要求 值得注意的是，这使我们能够创建一个廉价的读者，任何研究实验室都可以 买得起。为了更轻松地采用等离子荧光增强的微阵列，我们有 还创建了用于微阵列数据分析的软件。 在这个项目中，我们的目标是：将我们的等离子体荧光的制造量表提早 商业规模；更广泛地验证各种流行抗体的增强 微阵列；最终确定廉价读者的设计；更充分地开发我们的用户 - 友好的微阵列分析软件。 我们认为，等离荧光将成为所有人的标准荧光报告分子 微阵列和等离氟增强的微阵列将成为广泛使用且功能强大的 阐明蛋白质网络在健康和疾病中的作用的工具。

项目成果

Plasmonic Fluor-Enhanced Antigen Arrays for High-Throughput, Serological Studies of SARS-CoV-2.

• DOI: 10.1021/acsinfecdis.2c00086
• 发表时间: 2022-08-12
• 期刊: ACS INFECTIOUS DISEASES
• 影响因子: 5.3
• 作者: Qavi, Abraham J.;Wu, Chao;Lloyd, Matthew;Zaman, Mohammad Mahabub-Uz;Luan, Jingyi;Ballman, Claire;Leung, Daisy W.;Crick, Scott L.;Farnsworth, Christopher W.;Amarasinghe, Gaya K.
• 通讯作者: Amarasinghe, Gaya K.