Show simple item record

dc.contributor.author Vaidyanathan, Seetharaman
dc.contributor.author Kell, Douglas B.
dc.contributor.author Goodacre, Royston
dc.contributor.author Winder, Catherine L.
dc.contributor.author Wade, Steve C.
dc.date.accessioned 2009-12-21T10:23:40Z
dc.date.available 2009-12-21T10:23:40Z
dc.date.issued 2002-07-15
dc.identifier.citation Vaidyanathan , S , Kell , D B , Goodacre , R , Winder , C L & Wade , S C 2002 , ' Sample preparation in matrix-assisted laser desorption/ionization mass spectrometry of whole bacterial cells and the detection of high mass ( >20 kDa) proteins ' Rapid Communications in Mass Spectrometry , vol 16 , no. 13 , pp. 1276-1286 . , 10.1002/rcm.713 en
dc.identifier.issn 0951-4198
dc.identifier.other PURE: 133511
dc.identifier.other dspace: 2160/3873
dc.identifier.uri http://hdl.handle.net/2160/3873
dc.description Vaidyanathan, S., Winder, C. L., Wade, S. C., Kell, D. B., Goodacre, R. (2002). Sample preparation in matrix-assisted laser desorption/ionization mass spectrometry of whole bacterial cells and the detection of high mass (>20 kDa) proteins. Rapid Communications in Mass Spectrometry, 16, (13), 1276-1286 Sponsorship: BBSRC en
dc.description.abstract Three sample preparation strategies commonly employed in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOFMS) of whole bacterial cells were investigated for the detection of high mass signals; these included the dried droplet, the seed-layer/two-layer, and the bottom-layer methods. Different sample preparation approaches favoured the detection of high- or low-mass proteins. The low-mass peaks were best detected using the bottom-layer method. By contrast, the dried droplet method using a solvent with higher water content, and hence effecting a slower crystallization process, gave the best results for the detection of high-mass signals. Signals up to m/z 158 000 could be detected with this methodology for Bacillus sphaericus. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the same extracts used for MALDI-TOFMS showed bands in the molecular weight range in which high-mass peaks were observed in MALDI-MS, suggesting that the high-mass signals are not polymeric adducts of low-mass protein monomers. In addition, one of the high molecular weight proteins (126 kDa) was putatively identified as an S-layer protein by an in-gel tryptic digest. The bacterial samples spotted on the target wells for MALDI-TOFMS, using the different sample preparation strategies, were examined under a scanning electron microscope and differences were observed between the different strategies, suggesting that the nature of the crystals and the distribution of the analytes amidst the crystals could influence the spectral pattern observed in MALDI-TOFMS of whole bacterial cells. Finally, evidence is presented to indicate that, although the determinands are intact cells, cell lysis occurs both before and during the MALDI process. en
dc.format.extent 11 en
dc.language.iso eng
dc.relation.ispartof Rapid Communications in Mass Spectrometry en
dc.title Sample preparation in matrix-assisted laser desorption/ionization mass spectrometry of whole bacterial cells and the detection of high mass (>20 kDa) proteins en
dc.type Text en
dc.type.publicationtype Article (Journal) en
dc.identifier.doi http://dx.doi.org/10.1002/rcm.713
dc.contributor.institution Aberystwyth University en
dc.contributor.institution Institute of Biological, Environmental and Rural Sciences en
dc.description.status Peer reviewed en


Files in this item

Files Size Format View

There are no files associated with this item.

This item appears in the following Collection(s)

Show simple item record

Search Cadair


Advanced Search

Browse

Statistics