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用于蛋白质微阵列的等离子体纳米织构化 PMMA 表面:增加蛋白质结合和提高检测灵敏度。

Plasma nanotextured PMMA surfaces for protein arrays: increased protein binding and enhanced detection sensitivity.

机构信息

Institute of Microelectronics, NCSR Demokritos, P.O. Box 60228, Aghia Paraskevi, Attiki, 153 10 Greece.

出版信息

Langmuir. 2010 Sep 7;26(17):13883-91. doi: 10.1021/la101957w.

Abstract

Poly(methyl methacrylate) (PMMA) substrates were nanotextured through treatment in oxygen plasma to create substrates with increased surface area for protein microarray applications. Conditions of plasma treatment were found for maximum uniform protein adsorption on these nanotextured PMMA surfaces. Similar results were obtained using both a high-density plasma (HDP) and a low-density reactive ion etcher (RIE), suggesting independence from the plasma reactor type. The protein binding was evaluated by studying the adsorption of two model proteins, namely, biotinylated bovine serum albumin (b-BSA) and rabbit gamma-globulins (RgG). The immobilization of these proteins onto the surfaces was quantitatively determined through reaction with fluorescently labeled binding molecules. It was found that the adsorption of both proteins was increased up to 6-fold with plasma treatment compared to untreated surfaces and up to 4-fold compared to epoxy-coated glass slides. The sensitivity of detection was improved by 2 orders of magnitude. Moreover, highly homogeneous protein spots were created on optimized plasma-nanotextured surfaces through deposition with an automated microarray spotter, revealing the potential of plasma-nanotextured surfaces as protein microarray substrates.

摘要

聚甲基丙烯酸甲酯(PMMA)基底通过氧等离子体处理进行纳米纹理化,以在蛋白质微阵列应用中创建具有更大表面积的基底。找到了用于在这些纳米纹理化 PMMA 表面上实现最大均匀蛋白质吸附的等离子体处理条件。使用高密度等离子体(HDP)和低浓度反应离子蚀刻器(RIE)都获得了相似的结果,这表明与等离子体反应器类型无关。通过研究两种模型蛋白质,即生物素化牛血清白蛋白(b-BSA)和兔γ球蛋白(RgG)的吸附,评估了蛋白质结合。通过与荧光标记的结合分子反应,定量确定了这些蛋白质在表面上的固定化。与未处理的表面相比,等离子体处理将两种蛋白质的吸附增加了 6 倍,与环氧涂层玻璃载玻片相比增加了 4 倍。检测灵敏度提高了 2 个数量级。此外,通过自动化微阵列点样器在优化的等离子体纳米纹理化表面上沉积,创建了高度均匀的蛋白质斑点,显示出等离子体纳米纹理化表面作为蛋白质微阵列基底的潜力。

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