Free-flow
electrophoresis is a tool for the continuous fractionation
of electrically charged analytes. In this study, we introduce a novel
method to couple microchip-based free-flow electrophoresis with mass
spectrometry. The successive connection of multiple microchip outlets
to the electrospray ionization source of a mass spectrometer is automated
using a multiposition valve. With this novel setup, it is possible
to continuously fractionate and collect compounds while simultaneously
monitoring the process online with mass spectrometry. The functionality
of the method is demonstrated by the successful separation and identification
of the biomolecules AMP, ATP, and CoA, which are fundamental for numerous
biochemical processes in every organism.
Free-flow electrophoresis is
a tool for the continuous fractionation of electrically charged
analytes. In this study we introduce a novel method to couple
microchip-based free-flow electrophoresis with mass spectrometry. The
successive connection of multiple microchip outlets to the electrospray
ionization source of a mass spectrometer is automated using a
multiposition valve. With this novel setup it is possible to
continuously fractionate and collect compounds while simultaneously
monitoring the process online with mass spectrometry. The functionality
of the method is demonstrated by the successful separation and
identification of the biomolecules AMP, ATP and CoA, which are
fundamental for numerous biochemical processes in every organism.
Cell-free biosynthesis is emerging as a very attractive alternative for the production of market-relevant molecules. The free combination of enzymes, regardless of where they are isolated from, raises the possibility to build more efficient synthetic routes but at the same time leads to higher complexity regarding the analysis of the different enzymatic steps. Here we present an analytical method for the real-time analysis of acyl-CoA blocks forming and consuming during multi-step catalyses. We focused on malonyl-Coenzyme A and acetyl-CoA, which are the most used acyl-CoA units for carbon chain elongations. By employing capillary electrophoresis, we could detect the decrease of educts and the formation of products in a time-resolved fashion.
Free-flow electrophoresis is
a tool for the continuous fractionation of electrically charged
analytes. In this study we introduce a novel method to couple
microchip-based free-flow electrophoresis with mass spectrometry. The
successive connection of multiple microchip outlets to the electrospray
ionization source of a mass spectrometer is automated using a
multiposition valve. With this novel setup it is possible to
continuously fractionate and collect compounds while simultaneously
monitoring the process online with mass spectrometry. The functionality
of the method is demonstrated by the successful separation and
identification of the biomolecules AMP, ATP and CoA, which are
fundamental for numerous biochemical processes in every organism.
Free-flow electrophoresis is
a tool for the continuous fractionation of electrically charged
analytes. In this study we introduce a novel method to couple
microchip-based free-flow electrophoresis with mass spectrometry. The
successive connection of multiple microchip outlets to the electrospray
ionization source of a mass spectrometer is automated using a
multiposition valve. With this novel setup it is possible to
continuously fractionate and collect compounds while simultaneously
monitoring the process online with mass spectrometry. The functionality
of the method is demonstrated by the successful separation and
identification of the biomolecules AMP, ATP and CoA, which are
fundamental for numerous biochemical processes in every organism.
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