Electrochemical protein cleavage in a microfluidic cell with integrated boron doped diamond electrodes

Floris Teunis Gerardus van den Brink, Tao Zhang, Liwei Ma, Johan G. Bomer, Mathieu Odijk, Wouter Olthuis, Hjalmar P. Permentier, Rainer Bischoff, Albert van den Berg

    Research output: Contribution to journalArticleAcademicpeer-review

    9 Citations (Scopus)

    Abstract

    Specific electrochemical cleavage of peptide bonds at the C-terminal side of tyrosine and tryptophan generates peptides amenable to liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis for protein identification. To this end we developed a microfluidic electrochemical cell of 160 nL volume that combines a cell geometry optimized for a high electrochemical conversion efficiency (>95%) with an integrated boron doped diamond (BDD) working electrode offering a wide potential window in aqueous solution and reduced adsorption of peptides and proteins. Efficient cleavage of the proteins bovine insulin and chicken egg white lysozyme was observed at 4 out of 4 and 7 out of 9 of the predicted cleavage sites, respectively. Chicken egg white lysozyme was identified based on 5 electrochemically generated peptides using a proteomics database searching algorithm. These results show that electrochemical peptide bond cleavage in a microfluidic cell is a novel, fully instrumental approach toward protein analysis and eventually proteomics studies in conjunction with mass spectrometry
    Original languageEnglish
    Pages (from-to)9190-9198
    Number of pages9
    JournalAnalytical chemistry
    Volume88
    Issue number18
    DOIs
    Publication statusPublished - 20 Sep 2016

    Keywords

    • EWI-27326
    • IR-101825
    • METIS-318560

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