Using HPTLC/DESI-MS for peptide identification in 1D separations of tryptic protein digests

Sofie P. Pasilis; Vilmos Kertesz; Gary J. Van Berkel; Michael Schulz; Susanne Schorcht

doi:10.1007/s00216-008-1874-6

Using HPTLC/DESI-MS for peptide identification in 1D separations of tryptic protein digests

Sofie P. Pasilis, Vilmos Kertesz, Gary J. Van Berkel, Michael Schulz, Susanne Schorcht

Bioanalytical Mass Spectrometry

Research output: Contribution to journal › Article › peer-review

48 Scopus citations

Abstract

Desorption electrospray ionization mass spectrometry (DESI-MS) was investigated as a method to detect and identify peptides from tryptic digests of cytochrome c and myoglobin separated on ProteoChrom® HPTLC Silica gel 60 F_254s plates and ProteoChrom® HPTLC Cellulose sheets. Full-scan mass spectra and data-dependent tandem mass spectra were acquired in separate plate scans and used to identify peptide ions. Peptide distributions along the development lane were mapped for each separated protein digest. Signal levels ranged over several orders of magnitude. In general, highest signal levels were obtained for the peptides with the highest R _f values on a plate, while peptides with very low R _f values were often not detected. Sequence coverages for cytochrome c were 58% for the digest separated on the silica gel plate and 72% for the separation on the cellulose sheet; myoglobin sequence coverages were 62% and 68% on silica gel and cellulose, respectively. Weak correlations between peptide hydrophilicity and R _f values on the silica gel and cellulose plates were found, with the more hydrophilic peptides having lower R _f values.

Original language	English
Pages (from-to)	317-324
Number of pages	8
Journal	Analytical and Bioanalytical Chemistry
Volume	391
Issue number	1
DOIs	https://doi.org/10.1007/s00216-008-1874-6
State	Published - May 2008

Funding

Acknowledgements S.P.P. acknowledges an Oak Ridge National Laboratory (ORNL) appointment through the ORNL Postdoctoral Research Associates Program. Dr. Julian Philips (Thermo Fisher Scientific) is thanked for the loan of the LCQ DECA mass spectrometer. The MicroIonSpray head used to fabricate the DESI emitter was provided through a cooperative research and development agreement (CRADA) with MDS Sciex (ORNL02-0662). This research was supported by the Battelle Memorial Institute Technology Maturation Fund. The surface scanning platform and associated control software used in this study was developed with support from ORNL Technology Transfer and Economic Development (TTED) Royalty Funds. ORNL is managed and operated by UT-Battelle, LLC, for the United States Department of Energy under contract DE-AC05-00OR22725.

Funders	Funder number
Battelle Memorial Institute Technology Maturation Fund
Oak Ridge National Laboratory

Keywords

HPTLC/DESI-MS
Mass spectrometry
Peptides

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abstract = "Desorption electrospray ionization mass spectrometry (DESI-MS) was investigated as a method to detect and identify peptides from tryptic digests of cytochrome c and myoglobin separated on ProteoChrom{\textregistered} HPTLC Silica gel 60 F254s plates and ProteoChrom{\textregistered} HPTLC Cellulose sheets. Full-scan mass spectra and data-dependent tandem mass spectra were acquired in separate plate scans and used to identify peptide ions. Peptide distributions along the development lane were mapped for each separated protein digest. Signal levels ranged over several orders of magnitude. In general, highest signal levels were obtained for the peptides with the highest R f values on a plate, while peptides with very low R f values were often not detected. Sequence coverages for cytochrome c were 58% for the digest separated on the silica gel plate and 72% for the separation on the cellulose sheet; myoglobin sequence coverages were 62% and 68% on silica gel and cellulose, respectively. Weak correlations between peptide hydrophilicity and R f values on the silica gel and cellulose plates were found, with the more hydrophilic peptides having lower R f values.",

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AU - Kertesz, Vilmos

AU - Van Berkel, Gary J.

AU - Schulz, Michael

AU - Schorcht, Susanne

PY - 2008/5

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Using HPTLC/DESI-MS for peptide identification in 1D separations of tryptic protein digests

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