Noncontact 3D Bioprinting of Proteinaceous Microarrays for Highly Sensitive Immunofluorescence Detection within Clinical Samples.

IF 15.8 1区 材料科学 Q1 CHEMISTRY, MULTIDISCIPLINARY ACS Nano Pub Date : 2024-11-12 Epub Date: 2024-10-29 DOI:10.1021/acsnano.4c12460
Amid Shakeri, Lubna Najm, Shadman Khan, Lei Tian, Liane Ladouceur, Hareet Sidhu, Nadine Al-Jabouri, Zeinab Hosseinidoust, Tohid F Didar
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Abstract

Immunofluorescence assays are extensively used for the detection of disease-associated biomarkers within patient samples for direct diagnosis. Unfortunately, these 2D microarrays suffer from low repeatability and fail to attain the low limits of detection (LODs) required to accurately discern disease progression for clinical monitoring. While three-dimensional microarrays with increased biorecognition molecule density stand to circumvent these limitations, their viscous component materials are not compatible with current microarray fabrication protocols. Herein, we introduce a platform for 3D microarray bioprinting, wherein a two-step printing approach enables the high-throughput fabrication of immunosorbent hydrogels. The hydrogels are composed entirely of cross-linked proteins decorated with clinically relevant capture antibodies. Compared to two-dimensional microarrays, these proteinaceous microarrays offer 3-fold increases in signal intensity. When tested with clinically relevant biomarkers, ultrasensitive single-plex and multiplex detection of interleukin-6 (LOD 0.3 pg/mL) and tumor necrosis factor receptor 1 (LOD 1 pg/mL) is observed. When challenged with clinical samples, these hydrogel microarrays consistently discern elevated levels of interleukin-6 in blood plasma derived from patients with systemic blood infections. Given their easy-to-implement, high-throughput fabrication, and ultrasensitive detection, these three-dimensional microarrays will enable better clinical monitoring of disease progression, yielding improved patient outcomes.

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用于临床样本高灵敏度免疫荧光检测的蛋白质微阵列非接触式三维生物打印技术。
免疫荧光测定法被广泛用于检测患者样本中与疾病相关的生物标记物,以直接进行诊断。遗憾的是,这些二维微阵列的可重复性较低,无法达到临床监测所需的低检测限(LOD),无法准确判别疾病进展。虽然提高了生物识别分子密度的三维微阵列可以规避这些限制,但其粘性成分材料与目前的微阵列制造协议不兼容。在此,我们介绍一种三维微阵列生物打印平台,通过两步打印法实现免疫吸附水凝胶的高通量制造。这些水凝胶完全由交联蛋白质组成,并饰有临床相关的捕获抗体。与二维微阵列相比,这些蛋白质微阵列的信号强度提高了 3 倍。用临床相关生物标记物进行测试时,可观察到对白细胞介素-6(LOD 0.3 pg/mL)和肿瘤坏死因子受体 1(LOD 1 pg/mL)的超灵敏单倍和多重检测。在检测临床样本时,这些水凝胶微阵列能持续发现全身性血液感染患者血浆中白细胞介素-6 水平的升高。鉴于这些三维微阵列易于实施、高通量制造和超灵敏检测,它们将能更好地对疾病进展进行临床监测,从而改善患者的预后。
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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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