Performance of five different electrospray ionisation sources in conjunction with rapid monolithic column liquid chromatography and fast MS/MS scanning

Burges, K.E.V., Lainson, A., Imrie, L., Fraser-Pitt, D., Yaga, R., Smith, D.G.E., Swart, R., Pitt, A.R. and Inglis, N.F. (2009) Performance of five different electrospray ionisation sources in conjunction with rapid monolithic column liquid chromatography and fast MS/MS scanning. Proteomics, 9(6), pp. 1720-1726. (doi:10.1002/pmic.200800200)

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Publisher's URL: http://dx.doi.org/10.1002/pmic.200800200

Abstract

The performances of five different ESI sources coupled to a polystyrene-divinylbenzene monolithic column were compared in a series of LC-ESI-MS/MS analyses of Escherichia coli outer membrane proteins. The sources selected for comparison included two different modifications of the standard electrospray source, a commercial low-flow sprayer, a stainless steel nanospray needle and a coated glass Picotip. Respective performances were judged on sensitivity and the number and reproducibility of significant protein identifications obtained through the analysis of multiple identical samples. Data quality varied between that of a ground silica capillary, with 160 total protein identifications, the lowest number of high quality peptide hits obtained (3012), and generally peaks of lower intensity; and a stainless steel nanospray needle, which resulted in increased precursor ion abundance, the highest-quality peptide fragmentation spectra (5414) and greatest number of total protein identifications (259) exhibiting the highest MASCOT scores (average increase in score of 27.5% per identified protein). The data presented show that, despite increased variability in comparative ion intensity, the stainless steel nanospray needle provides the highest overall sensitivity. However, the resulting data were less reproducible in terms of proteins identified in complex mixtures - arguably due to an increased number of high intensity precursor ion candidates.

Item Type:Articles
Keywords:Electrospray ionisation • Liquid chromatography-tandem mass spectrometry, Membrane proteins, Objective evaluation
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Pitt, Dr Andrew and Smith, Professor David
Authors: Burges, K.E.V., Lainson, A., Imrie, L., Fraser-Pitt, D., Yaga, R., Smith, D.G.E., Swart, R., Pitt, A.R., and Inglis, N.F.
Subjects:Q Science > QH Natural history > QH345 Biochemistry
Q Science > QH Natural history > QH301 Biology
College/School:College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Proteomics
ISSN:1615-9853

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