期刊
JOURNAL OF MASS SPECTROMETRY
卷 43, 期 12, 页码 1627-1635出版社
WILEY
DOI: 10.1002/jms.1431
关键词
DESI; peptides; HPTLC; mass spectrometry; imaging
资金
- Cooperative Research and Development Agreement (CRADA) [ORNL02-0662]
- ORNL Technology Transfer and Economic Development (TTED)
- Division of Chemical Sciences, Geosciences and Biosciences, United States Department of Energy
- Office of Basic Energy Sciences
- Battelle Memorial Institute Technology Maturation Fund
- United States Department of Energy [DE-AC05-00OR22725]
Desorption electrospray ionization mass spectrometry (DESI-MS) was demonstrated as a method to detect and identify peptides from two-dimensional separations of cytochrome c and myoglobin tryptic digests on ProteoChrom HPTLC Cellulose sheets. Data-dependent tandem mass spectra were acquired during lane scans across the TLC plates. Peptides and the corresponding proteins were identified using a protein database search software. Two-dimensional distributions of identified peptides were mapped for each separated protein digest. Sequence coverages for cytochrome c and myoglobin were 81 and 74%, respectively. These compared well with those determined using the more standard HPLC/ESI-MS/MS approach (89 and 84%, respectively). Preliminary results show that use of more sensitive instrumentation has the potential for improved detection of peptides with low R-f values and improvement in sequence coverage. However, less multiple charging and more sodiation were seen in HPTLC/DESI-MS spectra relative to HPLC/ESI-MS spectra, which can affect peptide identification by MS/MS. Methods to increase multiple charging and reduce the extent of sodiation are currently under investigation. Published in 2008 by John Wiley & Sons, Ltd.
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