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Microfluidic platform with mass spectrometry detection for the analysis of phosphoproteins
Author(s) -
Dawoud Abdulilah A.,
Sarvaiya Hetal A.,
Lazar Iulia M.
Publication year - 2007
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200700355
Subject(s) - microfluidics , instrumentation (computer programming) , mass spectrometry , bioanalysis , chromatography , dephosphorylation , throughput , sample preparation , chemistry , nanotechnology , computer science , analytical chemistry (journal) , materials science , phosphorylation , telecommunications , biochemistry , phosphatase , wireless , operating system
Abstract The development of novel and reliable technologies for the analysis of proteins and their post‐translational modifications, in particular, has recently received much attention and interest. The implementation of a fully integrated microfluidic device interfaced with MS detection for the analysis of phosphoproteins is presented in this paper. The microfluidic platform (3′′×1.5′′) comprises two individual sample processing systems: one for performing direct sample infusion and one for performing microfluidic LC separations. Various MS detection strategies, specific for the study of post‐translational modifications, were conducted using α‐casein as a model protein. Neutral loss ion mapping, data‐dependent triple‐play and neutral loss analysis, and in situ dephosphorylation followed by LC separation and MS detection were performed. Consistent results in identifying phosphopeptides with conventional and microfluidic instrumentation have been obtained. Unlike with conventional instrumentation, however, the microfluidic device enabled the completion of each analysis from only a few microliters of sample, in ∼10–15 min, and on a bioanalytical platform that facilitates multiplexing and disposability, and thus high‐throughput, contamination‐free analysis.