Sensitivity-improved blocking agent-free fluorescence polarization assay through surface modification using polyethylene glycol,Analyst

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Sensitivity-improved blocking agent-free fluorescence polarization assay through surface modification using polyethylene glycol
Analyst ( IF 3.6 ) Pub Date : 2024-09-03 , DOI: 10.1039/d4an00569d
Hao Liu _{1,

2} , Mao Fukuyama ₁ , Yu Ogura ₁ , Motohiro Kasuya ₃ , Sho Onose ₄ , Ayuko Imai ₄ , Koji Shigemura ₄ , Manabu Tokeshi ₅ , Akihide Hibara ₆

Affiliation

Fluorescence polarization (FP) assays are widely used to quantify biomolecules, and their combination with microfluidic devices has the potential for application in onsite analysis. However, the hydrophobic surface of polydimethylsiloxane (PDMS)-based microfluidic devices and the amphiphilicity of the blocking agents can cause the nonspecific adsorption of biomolecules, which in turn reduces the sensitivity of the FP assay. To address this, we demonstrated an FP assay with improved sensitivity in microfluidic devices using a polyethylene glycol-based surface modification to avoid the use of blocking agents. We evaluated the effectiveness of the modification in inhibiting nonspecific protein adsorption and demonstrated the improved sensitivity of the FP immunoassay (FPIA). Our study addressed the lack of sensitivity of FP assays in microfluidic devices, particularly for the quantification of low-abundance analytes.

中文翻译：

通过使用聚乙二醇进行表面改性，提高灵敏度的无封闭剂荧光偏振测定

荧光偏振（FP）分析广泛用于量化生物分子，它们与微流控设备的结合具有在现场分析中应用的潜力。然而，基于聚二甲基硅氧烷（PDMS）的微流体装置的疏水表面和封闭剂的两亲性会导致生物分子的非特异性吸附，进而降低 FP 测定的灵敏度。为了解决这个问题，我们展示了一种 FP 测定法，它使用基于聚乙二醇的表面改性在微流体装置中具有更高的灵敏度，以避免使用封闭剂。我们评估了修饰在抑制非特异性蛋白质吸附方面的有效性，并证明了 FP 免疫测定（FPIA）的敏感性提高。我们的研究解决了 FP 分析在微流体设备中缺乏灵敏度的问题，特别是对于低丰度分析物的定量。

更新日期：2024-09-03

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