TY - JOUR
T1 - Electrochemical protein cleavage in a microfluidic cell with integrated boron doped diamond electrodes
AU - van den Brink, Floris Teunis Gerardus
AU - Zhang, Tao
AU - Ma, Liwei
AU - Bomer, Johan G.
AU - Odijk, Mathieu
AU - Olthuis, Wouter
AU - Permentier, Hjalmar P.
AU - Bischoff, Rainer
AU - van den Berg, Albert
PY - 2016/9/20
Y1 - 2016/9/20
N2 - 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
AB - 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
U2 - 10.1021/acs.analchem.6b02413
DO - 10.1021/acs.analchem.6b02413
M3 - Article
SN - 0003-2700
VL - 88
SP - 9190
EP - 9198
JO - Analytical chemistry
JF - Analytical chemistry
IS - 18
ER -