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On-Chip Spyhole Nanoelectrospray Ionization Mass Spectrometry for Sensitive Biomarker Detection in Small Volumes
Journal of the American Society for Mass Spectrometry ( IF 3.2 ) Pub Date : 2018-03-20 , DOI: 10.1007/s13361-018-1937-7
Xiaoqin Zhong 1 , Liang Qiao 2 , Géraldine Stauffer 1 , Baohong Liu 2 , Hubert H. Girault 1
Affiliation  

A polyimide microfluidic chip with a microhole emitter (Ø 10–12 μm) created on top of a microchannel by scanning laser ablation has been designed for nanoelectrospray ionization (spyhole-nanoESI) to couple microfluidics with mass spectrometry. The spyhole-nanoESI showed higher sensitivity compared to standard ESI and microESI from the end of the microchannel. The limits of detection (LOD) for peptide with the spyhole-nanoESI MS reached 50 pM, which was 600 times lower than that with standard ESI. The present microchip emitter allows the analysis of small volumes of samples. As an example, a small cell lung cancer biomarker, neuron-specific enolase (NSE), was detected by monitoring the transition of its unique peptide with the spyhole-nanoESI MS/MS. NSE at 0.2 nM could be well identified with a signal to noise ratio (S/N) of 50, and thereby its LOD was estimated to be 12 pM. The potential application of the spyhole-nanoESI MS/MS in cancer diagnosis was further demonstrated with the successful detection of 2 nM NSE from 1 μL of human serum. Before the detection, the serum sample spiked with NSE was first depleted with immune spin column, then desalted by centrifugal filter device, and finally digested by trypsin, without any other complicated preparation steps. The concentration matched the real condition of clinical samples. In addition, the microchips can be disposable to avoid any cross contamination. The present technique provides a highly efficient way to couple microfluidics with MS, which brings additional values to various microfluidics and MS-based analysis.

中文翻译:

片上Spyhole纳米电喷雾电离质谱用于小体积敏感生物标志物检测

通过扫描激光烧蚀在微通道顶部形成具有微孔发射器(Ø10–12μm)的聚酰亚胺微流控芯片,已设计用于纳米电喷雾电离(spyhole-nanoESI),以将微流控技术与质谱联用。从标准微通道末端开始,与标准ESI和microESI相比,spehole-nanoESI显示出更高的灵敏度。带有spyhole-nanoESI MS的肽的检测限(LOD)达到50 pM,比标准ESI的检测限低600倍。本微芯片发射器允许分析少量样品。例如,通过用spyhole-nanoESI MS / MS监测其独特肽的转变,可以检测到一种小细胞肺癌生物标记物,即神经元特异性烯醇化酶(NSE)。信噪比(S / N)为50时,可以很好地识别NSE为0.2 nM,因此其LOD估计为12 pM。成功地从1μL人血清中检测到2 nM NSE,进一步证明了spyhole-nanoESI MS / MS在癌症诊断中的潜在应用。在检测之前,先将掺有NSE的血清样品用免疫旋转柱消耗,然后用离心过滤器装置脱盐,最后用胰蛋白酶消化,无需任何其他复杂的制备步骤。浓度符合临床样品的实际情况。另外,微芯片可以是一次性的,以避免任何交叉污染。本技术提供了一种将微流体与MS耦合的高效方法,这为各种微流体和基于MS的分析带来了额外的价值。成功地从1μL人血清中检测到2 nM NSE,进一步证明了spyhole-nanoESI MS / MS在癌症诊断中的潜在应用。在检测之前,先用免疫旋转柱去除加标有NSE的血清样品,然后通过离心过滤装置脱盐,最后用胰蛋白酶消化,无需任何其他复杂的制备步骤。浓度符合临床样品的实际情况。另外,微芯片可以是一次性的,以避免任何交叉污染。本技术提供了一种将微流体与MS耦合的高效方法,这为各种微流体和基于MS的分析带来了额外的价值。成功地从1μL人血清中检测到2 nM NSE,进一步证明了spyhole-nanoESI MS / MS在癌症诊断中的潜在应用。在检测之前,先用免疫旋转柱去除加标有NSE的血清样品,然后通过离心过滤装置脱盐,最后用胰蛋白酶消化,无需任何其他复杂的制备步骤。浓度符合临床样品的实际情况。另外,微芯片可以是一次性的,以避免任何交叉污染。本技术提供了一种将微流体与MS耦合的高效方法,这为各种微流体和基于MS的分析带来了额外的价值。用NSE加标的血清样品首先用免疫旋转柱消耗,然后用离心过滤装置脱盐,最后用胰蛋白酶消化,无需任何其他复杂的制备步骤。浓度符合临床样品的实际情况。另外,微芯片可以是一次性的,以避免任何交叉污染。本技术提供了一种将微流体与MS耦合的高效方法,这为各种微流体和基于MS的分析带来了额外的价值。用NSE加标的血清样品首先用免疫旋转柱消耗,然后用离心过滤装置脱盐,最后用胰蛋白酶消化,无需任何其他复杂的制备步骤。浓度符合临床样品的实际情况。另外,微芯片可以是一次性的,以避免任何交叉污染。本技术提供了一种将微流体与MS耦合的高效方法,这为各种微流体和基于MS的分析带来了额外的价值。
更新日期:2018-06-16
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