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使用基于微悬臂梁的打印机生成微型平面重组抗体阵列。

Generation of miniaturized planar ecombinant antibody arrays using a microcantilever-based printer.

机构信息

Department of Immunotechnology, Lund University, Medicon Village, SE-22381 Lund, Sweden. CREATE Health, Lund University, Medicon Village, SE-22381 Lund, Sweden.

出版信息

Nanotechnology. 2014 Jul 11;25(27):275104. doi: 10.1088/0957-4484/25/27/275104. Epub 2014 Jun 24.

Abstract

Miniaturized (Ø 10 μm), multiplexed (>5-plex), and high-density (>100 000 spots cm(-2)) antibody arrays will play a key role in generating protein expression profiles in health and disease. However, producing such antibody arrays is challenging, and it is the type and range of available spotters which set the stage. This pilot study explored the use of a novel microspotting tool, Bioplume(TM)-consisting of an array of micromachined silicon cantilevers with integrated microfluidic channels-to produce miniaturized, multiplexed, and high-density planar recombinant antibody arrays for protein expression profiling which targets crude, directly labelled serum. The results demonstrated that 16-plex recombinant antibody arrays could be produced-based on miniaturized spot features (78.5 um(2), Ø 10 μm) at a 7-125-times increased spot density (250 000 spots cm(-2)), interfaced with a fluorescent-based read-out. This prototype platform was found to display adequate reproducibility (spot-to-spot) and an assay sensitivity in the pM range. The feasibility of the array platform for serum protein profiling was outlined.

摘要

微型化(Ø10μm)、多重化(>5 重)和高密度(>100000 个斑点/cm²)的抗体微阵列将在健康和疾病中的蛋白质表达谱生成方面发挥关键作用。然而,产生这样的抗体微阵列是具有挑战性的,可用的点样器的类型和范围决定了其性能。本初步研究探索了一种新型微点样工具的应用,即 Bioplume(TM)-由带有集成微流道的微加工硅悬臂阵列组成-用于生成针对粗制、直接标记血清的蛋白质表达谱分析的微型化、多重化和高密度平面重组抗体微阵列。结果表明,可以基于微型化的斑点特征(78.5μm²,Ø10μm)生产 16 重重组抗体微阵列,其斑点密度增加了 7-125 倍(250000 个斑点/cm²),并与基于荧光的读取方式相连接。该原型平台显示出足够的重现性(斑点间)和在 pM 范围内的检测灵敏度。概述了该阵列平台用于血清蛋白质分析的可行性。

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