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Proteomics, Protein Chemistry

Proteins/Peptides, Biopharmaceuticals, etc.

In proteomics and protein chemistry EC/MS is used for specific protein/peptide cleavage, reduction of disulfide bridges for the assignment of disulfide bonds in biopharmaceuticals, removal of sodium / potassium adducts also referred to as monovalent cation adduct removal (de-clustering) and for adduct formation such as drug-protein binding, etc.

The EC/MS method is generally faster, cleaner and without the need of any enzymes (e.g., enzyme free cleavage of peptide bond). Furthermore no harsh reducing agents are required causing ion suppression in electrospray ionization (ESI). The resulting products are analyzed either directly by infusion EC/MS or by on-line EC/LC/MS.

Recently the gentle electrochemical reduction of the S-S bonds has been successfully applied for the unfolding of larger proteins, replacing the harsh chemical reduction that often jeopardized the H/D exchange.

For more information on electrochemically assisted reactions in Protein Chemistry, read the following applications:


Application Notes

Application Notebook

Posters

  • 221_070_01 - Online EC MS A Powerful Tool for Fast Prediction of Phase I and II Drug Metabolism
  • 221_076_01 - Expanding Role of Electrochemistry Mass Spectrometry in Life Sciences
  • 221_077_01 - Novel method for efficient reduction of disulfide bonds in peptides and proteins prior MS detection
  • 221_078_01 - Identification of electrochemically synthesized metabolites
  • 221_079_01 - Insights into Nucleic Acids Oxidation by ElectrochemistryLiquid ChromatographyMass Spectrometry
  • 221_090_01 - Degradation of Xenobiotics and Binding to Organic Matter by OnLine EC MS
  • 221_105_01 - Controlled Reduction of Disulfide Bonds in Biopharmaceuticals Using an Electrochemical Reactor Cell online with LCMS Rolduc 2014
  • 221_106_01 - ElectrochemistryMS a Powerful Tool in Drug Metabolism Rolduc 2014
  • 221_107_01 - ElectrochemistryMS a Powerful Tool in Drug Metabolism ASMS 2014
  • 221_108_01 - Controlled Reduction of Disulfide Bonds in Proteins and Peptides Using an Electrochemical Reactor Cell in online LCECMS ASMS 2014
  • 221_115_01 - Controlled Reduction of Disulfide Bonds in Biopharmaceuticals ElCheMS2015
  • 221_116_01 - Enhanced Pharmaceutical Stability ElCheMS2015
  • 221_117_01 - Mimicking Drug Metabolism ElCheMS2015
  • 221_118_01 - Electrochem Reduction of Disulfide Bonds ASMS2015
  • 221_120_01 - ECHDX for the Analysis of Protein Therapeutics HOS2015
  • 221_226_01 - The Application of ElectrochemistryMS to Pharmaceutical Stability Testing and Degradant Synthesis


24251601 - Electrochemical reduction of disulfide-containing proteins for hydrogen/deuterium exchange monitored by mass spectrometry.
Mysling S, Salbo R, Ploug M, Jørgensen TJ.; Anal Chem. 2013 Nov 19. [Epub ahead of print]

24077854 - A novel electrochemical method for efficient reduction of disulfide bonds in peptides and proteins prior to MS detection.
Kraj A, Brouwer HJ, Reinhoud N, Chervet JP.; Anal Bioanal Chem. 2013 Sep 29. [Epub ahead of print]

24018861 - On-Line Electrochemical Reduction of Disulfide Bonds: Improved FTICR-CID and -ETD Coverage of Oxytocin and Hepcidin.
Nicolardi S, Giera M, Kooijman P, Kraj A, Chervet JP, Deelder AM, van der Burgt YE.; J Am Soc Mass Spectrom. 2013 Sep 10. [Epub ahead of print]

23763302 - Boron-doped diamond electrodes for the electrochemical oxidation and cleavage of peptides.
Roeser J, Alting NF, Permentier HP, Bruins AP, Bischoff R.; Anal Chem. 2013 Jul 16;85(14):6626-32. doi: 10.1021/ac303795c. Epub 2013 Jun 25.

23286982 - In chemico evaluation of skin metabolism: Investigation of eugenol and isoeugenol by electrochemistry coupled to liquid chromatography and mass spectrometry.
Melles D, Vielhaber T, Baumann A, Zazzeroni R, Karst U.; J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Jan 15;913-914:106-12. doi: 10.1016/j.jchromb.2012.12.004. Epub 2012 Dec 11.

22592985 - Electrochemistry/liquid chromatography/mass spectrometry to demonstrate irreversible binding of the skin allergen p-phenylenediamine to proteins.
Jahn S, Faber H, Zazzeroni R, Karst U.; Rapid Commun Mass Spectrom. 2012 Jun 30;26(12):1415-25. doi: 10.1002/rcm.6247.

22448817 - Electrochemistry-assisted top-down characterization of disulfide-containing proteins.
Zhang Y, Cui W, Zhang H, Dewald HD, Chen H.; Anal Chem. 2012 Apr 17;84(8):3838-42. doi: 10.1021/ac300106y. Epub 2012 Apr 4.

22237914 - Investigation of some biologically relevant redox reactions using electrochemical mass spectrometry interfaced by desorption electrospray ionization.
Lu M, Wolff C, Cui W, Chen H.; Anal Bioanal Chem. 2012 Apr;403(2):355-65. doi: 10.1007/s00216-011-5679-7. Epub 2012 Jan 12.

22227746 - Electrochemical oxidation and protein adduct formation of aniline: a liquid chromatography/mass spectrometry study.
Melles D, Vielhaber T, Baumann A, Zazzeroni R, Karst U.; Anal Bioanal Chem. 2012 Apr;403(2):377-84. doi: 10.1007/s00216-011-5673-0. Epub 2012 Jan 8.

21359310 - Coupling of liquid chromatography with mass spectrometry by desorption electrospray ionization (DESI).
Zhang Y, Yuan Z, Dewald HD, Chen H.; Chem Commun (Camb). 2011 Apr 14;47(14):4171-3. doi: 10.1039/c0cc05736c. Epub 2011 Feb 28.

21197958 - Online mass spectrometric analysis of proteins/peptides following electrolytic cleavage of disulfide bonds.
Zhang Y, Dewald HD, Chen H.; J Proteome Res. 2011 Mar 4;10(3):1293-304. doi: 10.1021/pr101053q. Epub 2011 Feb 14.

24300002 - Online Electrochemical Reduction of the Disulfide Bond(s) in Oxytocin and Hepcidin Results in Different CID and ETD Fragmentation Spectra
Martin A. Giera; Simone Nicolardi; Pieter Kooijman; Agnieszka Kraj; Jean-Pierre Chervet; Andre M. Deelder; Yuri E.M. van der Burgt; ASMS 2013 poster

See our support pages for a full list of recent literature