Using HPTLC/DESI-MS for Peptide Identification in 1D Separations of Tryptic Protein Digests

Journal Article · Tue Jan 01 04:00:00 EST 2008 · Analytical and Bioanalytical Chemistry

DOI:https://doi.org/10.1007/s00216-008-1874-6· OSTI ID:961556

Pasilis, Sofie P ^[1]; Kertesz, Vilmos ^[1]; Van Berkel, Gary J ^[1]; Schulz, Michael ^[2]; Schorcht, Susanne ^[2]

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1. ORNL
2. Merck Research Laboratories

Desorption electrospray ionization mass spectrometry 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 F254s plates and ProteoChrom HPTLC Cellulose sheets. Both 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 Rf values on a plate, while peptides with very low Rf 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 Rf values on the silica gel and cellulose plates were found, with more hydrophilic peptides having lower Rf values.

• Research Organization: Oak Ridge National Laboratory (ORNL)
• Sponsoring Organization: ORNL work for others
• DOE Contract Number: AC05-00OR22725
• OSTI ID: 961556
• Journal Information: Analytical and Bioanalytical Chemistry, Journal Name: Analytical and Bioanalytical Chemistry Journal Issue: 1 Vol. 391; ISSN 1618-2642
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CELLULOSE
CYTOCHROMES
DESORPTION
IONIZATION
MASS SPECTRA
MASS SPECTROSCOPY
MYOGLOBIN
PEPTIDES
PLATES
PROTEINS
SILICA GEL