title: Electrochemical Protein Cleavage in a Microfluidic Cell with Integrated Boron Doped Diamond Electrodes.
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.
authors: BIOS-Lab on a Chip Group, MESA+ Institute for Nanotechnology and MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente , 7500 AE Enschede, The Netherlands.
authors: Floris T G
authors: FT
authors: van den Brink
authors: Analytical Biochemistry and Interfaculty Mass Spectrometry Center, Department of Pharmacy, University of Groningen , 9713 AV Groningen, The Netherlands.
authors: Tao
authors: T
authors: Zhang
authors: BIOS-Lab on a Chip Group, MESA+ Institute for Nanotechnology and MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente , 7500 AE Enschede, The Netherlands.
authors: Liwei
authors: L
authors: Ma
authors: BIOS-Lab on a Chip Group, MESA+ Institute for Nanotechnology and MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente , 7500 AE Enschede, The Netherlands.
authors: Johan
authors: J
authors: Bomer
authors: BIOS-Lab on a Chip Group, MESA+ Institute for Nanotechnology and MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente , 7500 AE Enschede, The Netherlands.
authors: Mathieu
authors: M
authors: Odijk
authors: BIOS-Lab on a Chip Group, MESA+ Institute for Nanotechnology and MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente , 7500 AE Enschede, The Netherlands.
authors: Wouter
authors: W
authors: Olthuis
authors: Analytical Biochemistry and Interfaculty Mass Spectrometry Center, Department of Pharmacy, University of Groningen , 9713 AV Groningen, The Netherlands.
authors: Hjalmar P
authors: HP
authors: Permentier
authors: Analytical Biochemistry and Interfaculty Mass Spectrometry Center, Department of Pharmacy, University of Groningen , 9713 AV Groningen, The Netherlands.
authors: Rainer
authors: R
authors: Bischoff
authors: BIOS-Lab on a Chip Group, MESA+ Institute for Nanotechnology and MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente , 7500 AE Enschede, The Netherlands.
authors: Albert
authors: A
authors: van den Berg