Engineered self-assembling monolayers for label free detection of influenza nucleoprotein.
Engineered self-assembling monolayers for label free detection of influenza nucleoprotein.
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DOI：
10.1007/s10544-015-9951-z
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发表时间：
2015
影响因子：
2.8
通讯作者：
Lakey, Jeremy H.
中科院分区：
文献类型：
--
作者：
Le Brun, Anton P.;Soliakov, Andrei;Shah, Deepan S. H.;Holt, Stephen A.;McGill, Alison;Lakey, Jeremy H.
关键词：
将纳米技术集成到可用的设备中需要结合自下而上和自上而下的方法。通常，测量和控制这些不同成分的技术是完全不同的，因此能够原位分析纳米级成分的方法变得越来越重要。在这里，我们描述了一种利用自组装单层工程蛋白嵌合体在微电子设备上显示定向抗体（IgG）阵列的策略，用于无标签检测流感核蛋白。通过一系列物理技术，包括表面等离子体共振、石英晶体微平衡和中子反射法，表征了生物界面的结构和功能特性。这种方法的组合揭示了一个13.5 nm厚的工程单层，它(i)在金表面自组装，（ii）在确定的方向上以高亲和力捕获IgG， （iii）特异性识别甲型流感核蛋白。此外，我们还表明，这种非共价自组装结构可以通过原位化学交联使结合的IgG解离不可逆，而不影响IgG的功能。因此，这些方法可以详细描述从具有纳米尺寸的可溶性工程分子到展示工作流感传感器原理的阵列的转变。本文的在线版本（doi:10.1007/s10544-015-9951-z）包含补充材料，仅供授权用户使用。
Integrating nanotechnology into useable devices requires a combination of bottom up and top down methodology. Often the techniques to measure and control these different components are entirely different, so methods that can analyse the nanoscale component in situ are of increasing importance. Here we describe a strategy that employs a self-assembling monolayer of engineered protein chimeras to display an array of oriented antibodies (IgG) on a microelectronic device for the label free detection of influenza nucleoprotein. The structural and functional properties of the bio-interface were characterised by a range of physical techniques including surface plasmon resonance, quartz-crystal microbalance and neutron reflectometry. This combination of methods reveals a 13.5 nm thick engineered-monolayer that (i) self-assembles on gold surfaces, (ii) captures IgG with high affinity in a defined orientation and (iii) specifically recognises the influenza A nucleoprotein. Furthermore we also show that this non-covalent self-assembled structure can render the dissociation of bound IgG irreversible by chemical crosslinking in situ without affecting the IgG function. The methods can thus describe in detail the transition from soluble engineered molecules with nanometre dimensions to an array that demonstrates the principles of a working influenza sensor. The online version of this article (doi:10.1007/s10544-015-9951-z) contains supplementary material, which is available to authorized users.
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影响因子：
64.8
作者：
Cornell, BA;BraachMaksvytis, VLB;Pace, RJ
通讯作者：
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影响因子：
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通讯作者：
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