Evaluation of adsorption preconcentration/capillary zone electrophoresis/nanoelectrospray mass spectrometry for peptide and glycoprotein analyses

Bateman, Kevin P.; White, Robert L.; Thibault, Pierre

doi:10.1002/(sici)1096-9888(199811)33:11<1109::aid-jms728>3.0.co;2-1

Cited by 52 publications

(19 citation statements)

References 6 publications

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“…This is presumably due to incomplete desorption of analytes using a relatively low volume of organic buffer (25 nL) to minimize loss of separation performance [36]. Adsorptive preconcentration favored the enrichment of hydrophobic peptides whereas sample stacking provided significant enhancement of sensitivity for hydrophilic peptides of high electrophoretic mobilities.…”

Section: Discussionmentioning

confidence: 99%

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Rapid and sensitive separation of trace level protein digests using microfabricated devices coupled to a quadrupole - time-of-flight mass spectrometer

et al. 2000

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The application of microfabricated devices coupled to a quadrupole time‐of‐flight mass spectrometer (Qq‐TOF‐MS) is presented for the analysis of trace level digests of gel‐isolated proteins. In order to enhance the sample loading for proteomics analyses, two different on‐chip sample preconcentration techniques were evaluated. First, a sample stacking procedure that used polarity switching to remove the sample buffer prior to zone electrophoresis was easily integrated on the microfabricated devices. With the present chip design, this preconcentration technique provided up to 70 nL sample injection with sub‐nM detection limits for most peptide standards. For applications requiring larger sample loading, a disposable adsorption preconcentrator using a C18 membrane is incorporated outside the chip. This preconcentration method yielded lower peptide recoveries than that obtainable with sample stacking, and provided a convenient means of injecting several μL of sample with detection limits of typically 2.5 nM for hydrophobic peptides. The analytical merits of both sample enrichment approaches are described for the identification of bands isolated from two‐dimensional (2‐D) gel separation of protein extracts from Haemophilus influenzae. Accurate molecular mass measurements (< 5 ppm) in peptide mapping experiments is obtained by introducing an internal standard via a post‐separation channel. Rapid identification of trace level peptides is also demonstrated using on‐line tandem mass spectrometry and database searching with peptide sequence tags.

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Section: Discussionmentioning

confidence: 99%

“…Previous investigations have described the application of SPE to selectively enrich proteolytic fragments prior to CE/ nES-MS analyses [33,36]. Previous investigations have described the application of SPE to selectively enrich proteolytic fragments prior to CE/ nES-MS analyses [33,36].…”

Section: Spe Coupled To the Chip Ce/nes-msmentioning

confidence: 99%

Rapid and sensitive separation of trace level protein digests using microfabricated devices coupled to a quadrupole - time-of-flight mass spectrometer

et al. 2000

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“…Different approaches of on-line SPE preconcentration in the CE capillary, e.g., [63] have been developed. The SPE-CE-MS approach has especially been applied in the context of peptide analysis [33,[64][65][66]. The usefulness of online SPE-CE-MS for quantitative measurements has been shown by Waterval et al [67].…”

Section: Coating and Preconcentrationmentioning

confidence: 99%

Quantitation in capillary electrophoresis-mass spectrometry

2005

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Quantitation in capillary electrophoresis-mass spectrometryCE-MS has evolved into a strong alternative to LC-MS. Most of CE-MS applications deal with characterization and identification. However, quantitative aspects have gained importance in, e.g., pharmaceutical and biotechnological applications. Here we summarize and evaluate various methodological aspects in order to achieve sensitive and reproducible results. Similar to LC-MS, aspects of matrix influence on the electrospray process need to be carefully addressed when quantitative results are intended by CE-MS. Due to a more complicated coupling special emphasis needs to be put on the CE-MS interface. Generally linearity over more than three orders of magnitude can be achieved by CE-ESI-MS. Furthermore, a literature survey has been performed in order to give an overview over quantitative measurements performed by CE-MS. The precision can be doubled when changing from a structural related to an isotopically labeled internal standard. Thus a level of precision better than 5% RSD can be achieved.

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“…To improve sensitivity, more or less simple on-line enrichment can be applied [15][16][17][18]. Not only off-line but also on-capillary preconcentration by a stationary phase like C18-material on particles or absorbed on a membrane can be performed since CZE is very tolerant with respect to organic solvents [21][22][23]. Several groups (e.g., [19,20]) used transient-ITP to preconcentrate peptides at least by a factor of 10.…”

Section: Introductionmentioning

confidence: 99%

A robust approach for the analysis of peptides in the low femtomole range by capillary electrophoresis-tandem mass spectrometry

2002

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A capillary electrophoresis-tandem mass spectrometry (CE-MS/MS) approach has been developed for routine application in proteomic studies. Robustness of the coupling is achieved by using a standard coaxial sheath-flow sprayer. Thereby, greater stability than nanoelectrospray ionization-mass spectrometry coupling of sheathless capillary electrophoresis or nanoliquid chromatography (nano-LC) is achieved, resulting in stable operation for several weeks and unattended overnight sequences. The applied sheath flow is reduced to 1-2 microL/min in order to increase sensitivity. Standard peptides and those of digests of standard proteins and gel-separated proteins can be detected in the low femtomole range (full scan and MS/MS). Detection limits are found to be as low as 500 amol. Low femtomole amounts are required for unequivocal identification by MS/MS experiments in the ion trap and subsequent database search. By applying a simple pH-mediated stacking the concentration sensitivity can be lowered to some tens of fmol/microL (nM), depending on capillary size. This sensitivity is close to published values for sheathless CE-MS and nano-LC-MS, respectively (a comparison to reference values is presented). Moreover, with capillaries of about 50 cm in length separations in less than 10 min are possible resulting in a throughput of up to four analyses per hour. This is a factor of 4-12 times faster than nano-LC separation, being the state-of-the-art techniques for proteomic studies.

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Evaluation of adsorption preconcentration/capillary zone electrophoresis/nanoelectrospray mass spectrometry for peptide and glycoprotein analyses

Cited by 52 publications

References 6 publications

Rapid and sensitive separation of trace level protein digests using microfabricated devices coupled to a quadrupole - time-of-flight mass spectrometer

Rapid and sensitive separation of trace level protein digests using microfabricated devices coupled to a quadrupole - time-of-flight mass spectrometer

Quantitation in capillary electrophoresis-mass spectrometry

A robust approach for the analysis of peptides in the low femtomole range by capillary electrophoresis-tandem mass spectrometry

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