Analysis of peptides using <i>N</i>‐methylpolyvinylpyridium as silica surface modifier for CE‐ESI‐MS

Elhamili, Anisa; Wetterhall, Magnus; Sjödin, Marcus; Sebastiano, Roberto; Bergquist, Jonas

doi:10.1002/elps.200900536

Cited by 17 publications

(9 citation statements)

References 30 publications

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“…To overcome this disadvantage, a bare‐fused silica capillary and 10% (v/v) acetic acid (pH 2.3) was used as BGE for further separation studies. The lower pH of this BGE minimizes both, possible interactions of peptides with the capillary surface and the magnitude of the EOF, which is pH dependent and rises rapidly above pH = 4 . When using a bare‐fused silica capillary the capillary surface is negatively charged and the resulting EOF directed to the cathode.…”

Section: Resultsmentioning

confidence: 99%

Investigating capillary electrophoresis‐mass spectrometry for the analysis of common post‐translational modifications

et al. 2018

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Capillary electrophoresis coupled to mass spectrometry is a very efficient analytical method for the analysis of post‐translational modifications because of its high separation efficiency and high detection sensitivity. Here we applied CE‐MS using three differently coated separation capillaries for in‐depth analysis of a set of 70 synthetic post‐translationally modified peptides (including phosphorylation, acetylation, methylation, and nitration). We evaluated the results in terms of peptide detection and separation characteristics and found that the use of a neutrally coated capillary resulted in highest overall signal intensity of singly modified peptides. In contrast, the use of a bare‐fused silica capillary was superior in the identification of multi‐phosphorylated peptides (12 out of 15 were identified). Fast separations of approximately 12 min could be achieved using a positively coated capillary, however, at the cost of separation efficiency. A comparison to nanoLC‐MS revealed that multi‐phosphorylated peptides interact with the RP material very poorly so that these peptides were either washed out or elute as very broad peaks from the nano column which results in a reduced peptide identification rate (7 out of 15). Moreover, the methods applied were found to be very well suited for the analysis of the acetylated, nitrated and methylated peptides. All 36 synthetic peptides, which exhibit one of those modifications, could be identified regardless of the method applied. As a final step in this study and as a proof of principle, the phosphoproteome enriched from PC‐12 pheochromocytoma cells was analyzed by CE‐MS resulting in 5686 identified and 4088 quantified phosphopeptides. We compared the characterized analytes to those identified by a nanoLC‐MS proteomics study and found that less than one third of the phosphopeptides were identical, which demonstrates the benefit by combining different approaches quite impressively.

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Section: Resultsmentioning

confidence: 99%

Investigating capillary electrophoresis‐mass spectrometry for the analysis of common post‐translational modifications

et al. 2018

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“…An N ‐methylpolyvinylpyridinium cationic polymer‐based FS capillary coating, combined with an isoelectric, aspartic acid BGE, provided rapid, high‐efficient and high‐repeatable CE separations of both acidic and basic peptides and proteins with UV and/or ESI‐MS detection 87, 88. The importance of hydrophilic coating for CE analyses of peptides and proteins is demonstrated by their distorted/ruined separations when instead of N ‐methylpolyvinylpyridinium N ‐ethyl or N ‐octyl derivatives are used, respectively.…”

Section: Suppression Of Adsorption and Control Of Eofmentioning

confidence: 99%

Recent developments in CE and CEC of peptides (2009–2011)

Kašička

2011

Electrophoresis

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The review brings a comprehensive survey of the recent developments of high-performance electroseparation methods in capillary and microchip formats: zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography and electrochromatography. Applications of these techniques to analysis, isolation, purification and physicochemical and biochemical characterization of peptides are described. Advances in the investigation of electromigration properties of peptides, and in the methodology of their analysis, such as sample preparation, adsorption suppression, EOF control and detection, are presented. New developments, in particular, CE and CEC modes are reported and several types of their applications to peptide analysis are described: conventional qualitative and quantitative analysis, determination in complex (bio)matrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid, sequence and chiral analysis, and peptide mapping of proteins. Some micropreparative peptide separations are shown and capabilities of CE and CEC techniques to provide relevant physicochemical characteristics of peptides are demonstrated.

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“…Several types of internal capillary coating are described to reduce interaction of proteins and peptides with the capillary wall, as well as the electroosmotic flow. Those types of coatings and their potential advantages were described in detail in recent manuscripts and reviews 6–12. However, both phenomena appear to be of little or no consideration at the very acidic pH of 2–2.5 that is typically used in peptide separation.…”

Section: Technical Aspects Of Ce‐msmentioning

confidence: 99%

CE‐MS in biomarker discovery, validation, and clinical application

Mischak

Schanstra

2010

Proteomics Clinical Apps

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CE coupled MS (CE-MS) has become an increasingly employed technology in proteome analysis with focus on the identification of biomarker peptides in clinical proteomics. In this review, we will cover technical aspects of CE-MS coupling and highlight the improvements made in the last few years. We examine CE-MS from an application point of view, and evaluate its merits and vices for biomarker discovery and clinical applications. We discuss the principal theoretical and practical obstacles encountered when employing CE-MS (and most other proteomic technologies) for the analysis of body fluids for biomarker discovery. We will present several examples of a successful application of CE-MS for biomarker discovery, implications for disease diagnosis, prognosis, and therapy evaluation, and will discuss current challenges and possible future improvements.

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Analysis of peptides using N‐methylpolyvinylpyridium as silica surface modifier for CE‐ESI‐MS

Cited by 17 publications

References 30 publications

Investigating capillary electrophoresis‐mass spectrometry for the analysis of common post‐translational modifications

Investigating capillary electrophoresis‐mass spectrometry for the analysis of common post‐translational modifications

Recent developments in CE and CEC of peptides (2009–2011)

CE‐MS in biomarker discovery, validation, and clinical application

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