Chromatofocusing nonporous reversed-phase high-performance liquid chromatography/electrospray ionization time-of-flight mass spectrometry of proteins from human breast cancer whole cell lysates: a novel two-dimensional liquid chromatography/mass spectrometry method
dc.contributor.author | Chong, Bathsheba E. | en_US |
dc.contributor.author | Yan, Fang | en_US |
dc.contributor.author | Lubman, David M. | en_US |
dc.contributor.author | Miller, Fred R. | en_US |
dc.date.accessioned | 2006-04-19T14:08:13Z | |
dc.date.available | 2006-04-19T14:08:13Z | |
dc.date.issued | 2001-02-28 | en_US |
dc.identifier.citation | Chong, Bathsheba E.; Yan, Fang; Lubman, David M.; Miller, Fred R. (2001)."Chromatofocusing nonporous reversed-phase high-performance liquid chromatography/electrospray ionization time-of-flight mass spectrometry of proteins from human breast cancer whole cell lysates: a novel two-dimensional liquid chromatography/mass spectrometry method." Rapid Communications in Mass Spectrometry 15(4): 291-296. <http://hdl.handle.net/2027.42/35081> | en_US |
dc.identifier.issn | 0951-4198 | en_US |
dc.identifier.issn | 1097-0231 | en_US |
dc.identifier.uri | https://hdl.handle.net/2027.42/35081 | |
dc.identifier.uri | http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=11223961&dopt=citation | en_US |
dc.description.abstract | A novel two-dimensional two-column liquid chromatography/mass spectrometry (LC/MS) technique is described in this work, where chromatofocusing (CF) has been coupled to nonporous reversed-phase (NPS-RP) HPLC to separate proteins from human breast epithelial whole cell lysates. The liquid fractions from NPS-RP-HPLC are readily amenable to direct on-line analysis using electrospray ionization orthogonal acceleration time-of-flight mass spectrometry (ESI-TOFMS). A key advantage of this technique is that proteins can be ‘peeled off’ in the liquid phase from the CF column according to their isoelectric points ( pI ) in the first chromatographic separation dimension. The NPS-RP-HPLC column further separates these pI -focused fractions based upon protein hydrophobicity as the second chromatographic dimension. The third dimension involves on-line molecular weight determination using ESI-TOFMS. As a result, this method has the potential to be fully automated. In addition, a 2-D protein map of pI versus molecular weight is generated, which is analogous to a 2-D gel image. Thus, this technique may provide a means to study differential expression of proteins from whole cell lysates. Copyright © 2001 John Wiley & Sons, Ltd. | en_US |
dc.format.extent | 4252581 bytes | |
dc.format.extent | 3118 bytes | |
dc.format.mimetype | application/pdf | |
dc.format.mimetype | text/plain | |
dc.language.iso | en_US | |
dc.publisher | John Wiley & Sons, Ltd. | en_US |
dc.subject.other | Chemistry | en_US |
dc.subject.other | Analytical Chemistry and Spectroscopy | en_US |
dc.title | Chromatofocusing nonporous reversed-phase high-performance liquid chromatography/electrospray ionization time-of-flight mass spectrometry of proteins from human breast cancer whole cell lysates: a novel two-dimensional liquid chromatography/mass spectrometry method | en_US |
dc.type | Article | en_US |
dc.rights.robots | IndexNoFollow | en_US |
dc.subject.hlbtoplevel | Science | en_US |
dc.description.peerreviewed | Peer Reviewed | en_US |
dc.contributor.affiliationum | Department of Chemistry, The University of Michigan, Ann Arbor, MI 48109-1055, USA | en_US |
dc.contributor.affiliationum | Department of Chemistry, The University of Michigan, Ann Arbor, MI 48109-1055, USA | en_US |
dc.contributor.affiliationum | Department of Chemistry, The University of Michigan, Ann Arbor, MI 48109-1055, USA ; Department of Chemistry, The University of Michigan, Ann Arbor, MI 48109-1055, USA | en_US |
dc.contributor.affiliationother | Karmanos Cancer Institute, Wayne State University School of Medicine, Detroit, MI 48201, USA | en_US |
dc.identifier.pmid | 11223961 | en_US |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/35081/1/227_ftp.pdf | en_US |
dc.identifier.doi | http://dx.doi.org/10.1002/rcm.227 | en_US |
dc.identifier.source | Rapid Communications in Mass Spectrometry | en_US |
dc.owningcollname | Interdisciplinary and Peer-Reviewed |
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