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 journalArticle

  • 5 Citations

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
LanguageEnglish
Pages9190-9198
Number of pages9
JournalAnalytical chemistry
Volume88
Issue number18
DOIs
StatePublished - 20 Sep 2016

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Diamond
Boron
Microfluidics
Peptides
Electrodes
Muramidase
Proteins
Mass spectrometry
Electrochemical cells
Liquid chromatography
Tryptophan
Conversion efficiency
Tyrosine
Insulin
Adsorption
Geometry

Keywords

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

Cite this

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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",
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author = "{van den Brink}, {Floris Teunis Gerardus} and Tao Zhang and Liwei Ma and Bomer, {Johan G.} and Mathieu Odijk and Wouter Olthuis and Permentier, {Hjalmar P.} and Rainer Bischoff and {van den Berg}, Albert",
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doi = "10.1021/acs.analchem.6b02413",
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Electrochemical protein cleavage in a microfluidic cell with integrated boron doped diamond electrodes. / van den Brink, Floris Teunis Gerardus; Zhang, Tao; Ma, Liwei; Bomer, Johan G.; Odijk, Mathieu; Olthuis, Wouter; Permentier, Hjalmar P.; Bischoff, Rainer; van den Berg, Albert.

In: Analytical chemistry, Vol. 88, No. 18, 20.09.2016, p. 9190-9198.

Research output: Contribution to journalArticle

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

N1 - eemcs-eprint-27326

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

KW - EWI-27326

KW - IR-101825

KW - METIS-318560

U2 - 10.1021/acs.analchem.6b02413

DO - 10.1021/acs.analchem.6b02413

M3 - Article

VL - 88

SP - 9190

EP - 9198

JO - Analytical chemistry

T2 - Analytical chemistry

JF - Analytical chemistry

SN - 0003-2700

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ER -