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Issue 9, 2016
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Continuous on-chip fluorescence labelling, free-flow isoelectric focusing and marker-free isoelectric point determination of proteins and peptides

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Abstract

We present a microfluidic platform that contains a micro flow reactor for on-chip biomolecule labelling that is directly followed by a separation bed for continuous free-flow electrophoresis and has an integrated hydrogel-based near-infrared fluorescent pH sensor layer. Using this assembly, labelling of protein and peptide mixtures, their separation via free-flow isoelectric focusing and the determination of the isoelectric point (pI) of the separated products via the integrated sensor layer could be carried out within typically around 5 minutes. Spatially-resolved immobilization of fluidic and sensing structures was carried out via multistep photolithography. The assembly was characterized and optimized with respect to their fluidic and pH sensing properties and applied in the IEF of model proteins, peptides and a tryptic digest from physalaemine. We have therefore realized continuous sample preparation and preparative separation, analyte detection, process observation and analyte assignment capability based on pI on a single platform the size of a microscope slide.

Graphical abstract: Continuous on-chip fluorescence labelling, free-flow isoelectric focusing and marker-free isoelectric point determination of proteins and peptides

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Publication details

The article was received on 13 Jan 2016, accepted on 23 Mar 2016 and first published on 24 Mar 2016


Article type: Communication
DOI: 10.1039/C6LC00055J
Citation: Lab Chip, 2016,16, 1565-1572
  • Open access: Creative Commons BY-NC license
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    Continuous on-chip fluorescence labelling, free-flow isoelectric focusing and marker-free isoelectric point determination of proteins and peptides

    C. Herzog, E. Poehler, A. J. Peretzki, S. M. Borisov, D. Aigner, T. Mayr and S. Nagl, Lab Chip, 2016, 16, 1565
    DOI: 10.1039/C6LC00055J

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