Fluorinated liquid-enabled protein handling and surfactant-aided crystallization for fully in situ digital microfluidic MALDI-MS analysis

Lab on a Chip
Andrew P AijianRobin L Garrell

Abstract

A droplet (digital) microfluidic device has been developed that enables complete protein sample preparation for MALDI-MS analysis. Protein solution dispensing, disulfide bond reduction and alkylation, tryptic digestion, sample crystallization, and mass spectrometric analysis are all performed on a single device without the need for any ex situ sample purification. Fluorinated solvents are used as an alternative to surfactants to facilitate droplet movement and limit protein adsorption onto the device surface. The fluorinated solvent is removed by evaporation and so does not interfere with the MALDI-MS analysis. Adding a small amount of perfluorooctanoic acid to the MALDI matrix solution improves the yield, quality and consistency of the protein-matrix co-crystals, reducing the need for extensive 'sweet spot' searching and improving the spectral signal-to-noise ratio. These innovations are demonstrated in the complete processing and MALDI-MS analysis of lysozyme and cytochrome c. Because all of the sample processing steps and analysis can be performed on a single digital microfluidic device without the need for ex situ sample handling, higher throughput can be obtained in proteomics applications. More generally, the results pres...Continue Reading

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Citations

Apr 10, 2013·Analytical and Bioanalytical Chemistry·Yong Zeng, Tanyu Wang
Nov 13, 2012·Analytical Chemistry·Michelle L KovarikNancy L Allbritton
Sep 19, 2014·Current Opinion in Biotechnology·Amar OeditThomas Hankemeier
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Mar 4, 2014·Electrophoresis·Katrina N BattleSteven A Soper
Mar 31, 2016·Analytical Chemistry·Olena KudinaFrieder Mugele
Jan 9, 2014·Mass Spectrometry Reviews·Xiaojun FengXin Liu
Dec 18, 2013·Chemical Society Reviews·Josep Puigmartí-Luis
Jun 20, 2013·Analytical Chemistry·Andrea E Kirby, Aaron R Wheeler
Dec 17, 2014·Journal of Laboratory Automation·Andrew P Aijian, Robin L Garrell
Nov 27, 2020·Analytical Methods : Advancing Methods and Applications·Hee-Jin YooKun Cho

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