Coupling Droplet Microfluidics with Mass Spectrometry for Ultrahigh-Throughput Analysis of Complex Mixtures up to and above 30 Hz.
Coupling Droplet Microfluidics with Mass Spectrometry for Ultrahigh-Throughput Analysis of Complex Mixtures up to and above 30 Hz.
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DOI：
10.1021/acs.analchem.0c02632
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发表时间：
2020-09-15
影响因子：
7.4
通讯作者：
Barran PE
中科院分区：
文献类型：
--
作者：
Kempa EE;Smith CA;Li X;Bellina B;Richardson K;Pringle S;Galman JL;Turner NJ;Barran PE
从环境监测到药物发现和工业生物技术，许多应用都需要高通量和超高通量的无标记样品分析。HTS方法主要基于有针对性的工作流程，这可能会限制其范围。质谱法很容易提供复杂混合物的化学特性和丰度，在这里，我们使用微滴生成微流体技术以高达33 Hz的速率提供皮升等分试样进行分析。这是证明了小分子，肽和蛋白质高达66 kDa的三个市售的质谱仪从盐溶液模拟细胞环境。基于芯片的接口，允许这种耦合的设计，这些接口的优点和挑战进行了讨论。在Orbitrap平台上，6 Hz的液滴注入速率用于细胞色素c的分析，在DTIMS Q-TOF上获得类似的速率，并且在TWIMS Q-TOF上利用IM-MS软件，证明了高达33 Hz的速率。这种方法的潜力被证明与概念实验的证明，包括鸡蛋白色，未纯化的重组蛋白，和生物转化上清液的粗混合物。
High- and ultrahigh-throughput label-free sample analysis is required by many applications, extending from environmental monitoring to drug discovery and industrial biotechnology. HTS methods predominantly are based on a targeted workflow, which can limit their scope. Mass spectrometry readily provides chemical identity and abundance for complex mixtures, and here, we use microdroplet generation microfluidics to supply picoliter aliquots for analysis at rates up to and including 33 Hz. This is demonstrated for small molecules, peptides, and proteins up to 66 kDa on three commercially available mass spectrometers from salty solutions to mimic cellular environments. Designs for chip-based interfaces that permit this coupling are presented, and the merits and challenges of these interfaces are discussed. On an Orbitrap platform droplet infusion rates of 6 Hz are used for analysis of cytochrome c, on a DTIMS Q-TOF similar rates were obtained, and on a TWIMS Q-TOF utilizing IM-MS software rates up to 33 Hz are demonstrated. The potential of this approach is demonstrated with proof of concept experiments on crude mixtures including egg white, unpurified recombinant protein, and a biotransformation supernatant.
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影响因子：
16.6
作者：
Colin PY;Kintses B;Gielen F;Miton CM;Fischer G;Mohamed MF;Hyvönen M;Morgavi DP;Janssen DB;Hollfelder F
通讯作者：
Hollfelder F
影响因子：
--
作者：
Hodder, P;Mull, R;Strulovici, B
通讯作者：
Strulovici, B
影响因子：
7.4
作者：
He, MY;Edgar, JS;Chiu, DT
通讯作者：
Chiu, DT
DOI：
10.1146/annurev.anchem.1.031207.112945
发表时间：
2008-01-01
影响因子：
8
作者：
Marshall, Alan G.;Hendrickson, Christopher. L.
通讯作者：
Hendrickson, Christopher. L.
影响因子：
16.6
作者：
Hoffmann, Peter;Haeusig, Ulrich;Belder, Detlev
通讯作者：
Belder, Detlev