Performance of a sheathless porous tip sprayer for capillary electrophoresis–electrospray ionization-mass spectrometry of intact proteins

Haselberg, Rob; Ratnayake, Chitra; Jong, Gerhardus J. de; Somsen, Govert W.

doi:10.1016/j.chroma.2010.10.006

Cited by 94 publications

(88 citation statements)

References 48 publications

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“…Considering the results discussed above, we demonstrate, as in the case of CE-UV, limits of detection in the low nanomolar range by FESI-CE-MALDI-MS, which are comparable with detection limits of proteins achieved by state-of-art sheathless CE-ESI-MS [37]. …”

Section: Hyphenation Of Fesi-ce To Maldi-mssupporting

confidence: 74%

High‐sensitive protein analysis by FESI‐CE‐MALDI‐MS

2011

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Capillary zone electrophoresis (CZE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) are two techniques highly suitable for the separation and detection of intact proteins. Herein, based on the use of a recently introduced iontophoretic fraction collection interface for the coupling of CE and MALDI-MS, the potential of the combination of both techniques for the analysis of intact proteins is assessed. To further provide a bioanalytical platform with high-sensitivity capabilities, field-enhanced sample injection is integrated as on online preconcentration strategy upstream from the electrokinetic separation. Under optimized conditions, more than 3200-and 4800-fold improvement, respectively in terms of peak height and peak area, as well as LODs ranging from 5 to 10 nM, has been achieved.

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Section: Hyphenation Of Fesi-ce To Maldi-mssupporting

confidence: 74%

High‐sensitive protein analysis by FESI‐CE‐MALDI‐MS

2011

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“…39 -41. Applications such as the analysis of intact proteins (42), protein-protein and protein-metal complexes (43), and ribosomal protein digests from E. coli (44) have been published. Method-inherent advantages of CESI-MS are highly efficient separations, low flow rates leading to reduced ion suppression, and greater sensitivity (40).…”

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confidence: 99%

Comparing and Combining Capillary Electrophoresis Electrospray Ionization Mass Spectrometry and Nano–Liquid Chromatography Electrospray Ionization Mass Spectrometry for the Characterization of Post-translationally Modified Histones

Sarg

Faserl

Kremser

et al. 2013

Molecular & Cellular Proteomics

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We present the first comprehensive capillary electrophoresis electrospray ionization mass spectrometry (CESI-MS) analysis of post-translational modifications derived from H1 and core histones. Using a capillary electrophoresis system equipped with a sheathless high-sensitivity porous sprayer and nano-liquid chromatography electrospray ionization mass spectrometry (nano-LC-ESI-MS) as two complementary techniques, we characterized H1 histones isolated from rat testis. Without any pre-separation of the perchloric acid extraction, a total of 70 different modified peptides, including 50 phosphopeptides, were identified in the rat linker histones H1.0, H1a-H1e, and H1t. Out of the 70 modified H1 histone peptides, 27 peptides could be identified with CESI-MS only, and 11 solely with LC-ESI-MS. Immobilized metal-affinity chromatography enrichment prior to MS analysis yielded a total of 55 phosphopeptides; 22 of these peptides could be identified only by CESI-MS, and 19 only by LC-ESI-MS, showing the complementarity of the two techniques. We mapped 42 H1 modification sites, including 31 phosphorylation sites, of which 8 were novel sites. For the analysis of core histones, we chose a different strategy. In a first step, the sulfuric-acid-extracted core histones were pre-separated using reverse-phase high-performance liquid chromatography. Individual rat testis core histone fractions obtained in this way were digested and analyzed via bottom-up CESI-MS. This approach yielded the identification of 42 different modification sites including acetylation (lysine and N ␣ -terminal); mono-, di-, and trimethylation; and phosphorylation. When we applied CESI-MS for the analysis of intact core histone subtypes from butyrate-treated mouse tumor cells, we were able to rapidly detect their degree of modification, and we found this method very useful for the separation of isobaric trimethyl and acetyl modifications. Taken together, our results highlight the need for additional techniques for the comprehensive analysis of post-translational modifications. CESI-MS is a promising new proteomics tool as demonstrated by this, the first comprehensive analysis of histone modifications, using rat testis as an example. Molecular & Cellular

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“…A commercially available sheathless CESI source combines high resolution separation of CE with high resolution mass spectrometry. Because there is no sample dilution, as occurs in sheath-flow systems, the sheathless interface is more sensitive [235][236][237]. We [238] and others [229,239] have demonstrated the potential of high resolution CZE coupled to high resolution MS to analyze intact complex glycoproteins.…”

Section: Introductionmentioning

confidence: 99%

“…An alternative high resolution technique is capillary zone electrophoresis (CZE), which separates analytes based on differences in charge and hydrodynamic volume. CZE has been demonstrated to be a high efficiency separation method for the analysis of proteins [25,26,274,275]. In contrast to LC (except low resolution SEC), CZE does not involve interaction of the analyte with the stationary phase, thus representing a complementary separation approach.…”

Section: Introductionmentioning

confidence: 99%

“…However, since only the porous end of the separation capillary is housed in the reservoir, the passage of ions from the reservoir through the pores in the separation capillary is achieved without dilution of the BGE. A number of groups have utilized this interface in proteomics experiments [1, [66][67][68][69][70]. The interface is sold commercially via SCIEX (Toronto, Canada) and marketed for CZE-MS applications as a part of SCIEX's CESI platform.…”

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confidence: 99%

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Top-down proteomic analysis of protein pharmaceuticals, mixtures of protein complexes, and ribosomal protein extracts by capillary zone electrophoresis-mass spectrometry

Belov¹

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Performance of a sheathless porous tip sprayer for capillary electrophoresis–electrospray ionization-mass spectrometry of intact proteins

Cited by 94 publications

References 48 publications

High‐sensitive protein analysis by FESI‐CE‐MALDI‐MS

High‐sensitive protein analysis by FESI‐CE‐MALDI‐MS

Comparing and Combining Capillary Electrophoresis Electrospray Ionization Mass Spectrometry and Nano–Liquid Chromatography Electrospray Ionization Mass Spectrometry for the Characterization of Post-translationally Modified Histones

Top-down proteomic analysis of protein pharmaceuticals, mixtures of protein complexes, and ribosomal protein extracts by capillary zone electrophoresis-mass spectrometry

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