多重数字平台可检测阿托摩尔级蛋白质水平,交叉反应最小

IF 16 1区 材料科学 Q1 CHEMISTRY, MULTIDISCIPLINARY
Stephanie J. Zhang, Connie Wu and David R. Walt*, 
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引用次数: 0

摘要

基于蛋白质的生物标记物对疾病诊断至关重要,但它们在生物流体中的低丰度往往给传统的酶联免疫吸附测定(ELISA)技术带来巨大的检测挑战。虽然数字酶联免疫吸附分析等各种超灵敏方法提高了灵敏度,但多重检测仍存在相当大的交叉反应,从而影响检测结果的准确性。为了应对这一挑战,我们开发了条形码分子珠上信号放大个体计数法(barcoded MOSAIC),这是一种多重数字 ELISA 技术,通过将条形码检测抗体与特定类型的珠子配对,显著降低了交叉反应。这种方法能从不到 9 μL 的血液中同时检测八种分析物,灵敏度从中位数摩尔到低位数摩尔不等,单次多重检测中多种分析物的总动态范围超过七对数。此外,条形码 MOSAIC 与标准的免疫测定试剂和工作流程兼容,利用快速、自动的流式细胞仪读数进行定量,这使它成为一个非常容易使用的台式平台,研究和临床实验室均可采用,为将来转化为护理点应用奠定了基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

A Multiplexed Digital Platform Enables Detection of Attomolar Protein Levels with Minimal Cross-Reactivity

A Multiplexed Digital Platform Enables Detection of Attomolar Protein Levels with Minimal Cross-Reactivity

Protein-based biomarkers are essential for disease diagnostics, yet their low abundance in biofluids often presents significant detection challenges for traditional enzyme-linked immunosorbent assay (ELISA) techniques. While various ultrasensitive methods such as digital ELISA have improved sensitivity, multiplex assays still suffer from considerable cross-reactivities that can compromise result accuracies. To address this challenge, we have developed barcoded Molecular On-bead Signal Amplification for Individual Counting (barcoded MOSAIC), a multiplexed digital ELISA technology that markedly reduces cross-reactivity by pairing barcoded detection antibodies with specific bead types. This approach enables the simultaneous detection of eight analytes from less than 9 μL of blood, with sensitivities ranging from midpicomolar to low-attomolar levels and a collective dynamic range exceeding seven logs across multiple analytes within a single multiplex assay. Additionally, barcoded MOSAIC is compatible with standard immunoassay reagents and workflows, utilizing a rapid, automatable flow cytometric readout for quantification, which makes it a highly accessible benchtop platform that is readily adoptable by both research and clinical laboratories, setting the stage for future translation into point-of-care applications.

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来源期刊
ACS Nano
ACS Nano 工程技术-材料科学:综合
CiteScore
26.00
自引率
4.10%
发文量
1627
审稿时长
1.7 months
期刊介绍: ACS Nano, published monthly, serves as an international forum for comprehensive articles on nanoscience and nanotechnology research at the intersections of chemistry, biology, materials science, physics, and engineering. The journal fosters communication among scientists in these communities, facilitating collaboration, new research opportunities, and advancements through discoveries. ACS Nano covers synthesis, assembly, characterization, theory, and simulation of nanostructures, nanobiotechnology, nanofabrication, methods and tools for nanoscience and nanotechnology, and self- and directed-assembly. Alongside original research articles, it offers thorough reviews, perspectives on cutting-edge research, and discussions envisioning the future of nanoscience and nanotechnology.
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