Developments in Interfacing Designs for CE–MS: Towards Enabling Tools for Proteomics and Metabolomics

Lindenburg, Petrus W.; Haselberg, Rob; Rozing, Gerard P.; Ramautar, Rawi

doi:10.1007/s10337-014-2795-5

Cited by 67 publications

(42 citation statements)

References 77 publications

(121 reference statements)

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“…A microfluidic setting extended these electrophoretic studies to higher throughput, 12 erythrocytes/min (24). Continuous developments in CE separation (29,30) and lategeneration CE nano-flow ESI interfaces (31,32) (see reviews (33,34)) accomplished high-sensitivity detection of proteomes (35,36) and labile PTMs, such as phosphorylation (37). These developments enabled femtogram (zeptomole) limit of detection (38) for protein digests from cell populations.…”

mentioning

confidence: 99%

Label-free Quantification of Proteins in Single Embryonic Cells with Neural Fate in the Cleavage-Stage Frog (Xenopus laevis) Embryo using Capillary Electrophoresis Electrospray Ionization High-Resolution Mass Spectrometry (CE-ESI-HRMS)

Lombard‐Banek¹,

Reddy²,

Moody

et al. 2016

Molecular & Cellular Proteomics

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Quantification of protein expression in single cells promises to advance a systems-level understanding of normal development. Using a bottom-up proteomic workflow and multiplexing quantification by tandem mass tags, we recently demonstrated relative quantification between single embryonic cells (blastomeres) in the frog (Xenopus laevis) embryo. In this study, we minimize derivatization steps to enhance analytical sensitivity and use label-free quantification (LFQ) for single Xenopus cells. The technology builds on a custom-designed capillary electrophoresis microflow-electrospray ionization high-resolution mass spectrometry platform and LFQ by MaxLFQ (MaxQuant). By judiciously tailoring performance to peptide separation, ionization, and data-dependent acquisition, we demonstrate an ϳ75-amol (ϳ11 nM) lower limit of detection and quantification for proteins in complex cell digests. The platform enabled the identification of 438 nonredundant protein groups by measuring 16 ng of protein digest, or <0.2% of the total protein contained in a blastomere in the 16-cell embryo. LFQ intensity was validated as a quantitative proxy for protein abundance. Correlation analysis was performed to compare protein quantities between the embryo and n ‫؍‬ 3 different single D11 blastomeres, which are fated to develop into the nervous system. A total of 335 nonredundant protein groups were quantified in union between the single D11 cells spanning a 4 log-order concentration range. LFQ and correlation analysis detected expected proteomic differences between the whole embryo and blastomeres, and also found translational differences between individual D11 cells. LFQ on single cells raises exciting possibilities to study gene expression in other cells and models to help better understand cell processes on a systems biology level. Molecular & Cellular

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mentioning

confidence: 99%

Label-free Quantification of Proteins in Single Embryonic Cells with Neural Fate in the Cleavage-Stage Frog (Xenopus laevis) Embryo using Capillary Electrophoresis Electrospray Ionization High-Resolution Mass Spectrometry (CE-ESI-HRMS)

Lombard‐Banek¹,

Reddy²,

Moody

et al. 2016

Molecular & Cellular Proteomics

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“…There are currently two different interfaces commercially available, namely, the sheath-liquid and the porous tip sheathless interface [84]. The sheath-liquid interface is typically considered more stable, repeatable and cost effective but suffers from a lower sensitivity due to dilution of the CE effluent with the sheath liquid.…”

Section: Capillary Electrophoresismentioning

confidence: 99%

Analytical Pitfalls and Challenges in Clinical Metabolomics

et al. 2016

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Metabolomics-based strategies have become an integral part of modern clinical research, allowing for a better understanding of pathophysiological conditions and disease mechanisms, as well as providing innovative tools for more adequate diagnostic and prognosis approaches. Metabolomics is considered an essential tool in precision medicine, which aims for personalized prevention and tailor-made treatments. Nevertheless, multiple pitfalls may be encountered in clinical metabolomics during the entire workflow, hampering the quality of the data and, thus, the biological interpretation. This review describes the challenges underlying metabolomics-based experiments, discussing step by step the potential pitfalls of the analytical process, including study design, sample collection, storage, as well as preparation, chromatographic and electrophoretic separation, detection and data analysis. Moreover, it offers practical solutions and strategies to tackle these challenges, ensuring the generation of high-quality data.

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“…Recent reviews highlight the use of CZE and CE-ESI-MS for bottom-up proteomics, 18, 19, 143, 144 and CE and MCE methods for top-down proteomics including investigation of post-translational modifications (PTMs). 18 These reviews include advances that have been made for CE interfacing to ESI-MS, electrospray ionization-tandem mass spectrometry (ESI-MS/MS), and matrix-assisted laser desorption/ionization mass spectrometry (MADLI-MS).…”

Section: Applications Of Ce and Mcementioning

confidence: 99%

Recent advances in protein analysis by capillary and microchip electrophoresis

Dawod

Arvin

Kennedy

2017

Analyst

120

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This review article describes the significant recent advances in analysis of proteins by capillary and microchip electrophoresis during the period mid 2014 to early 2017. The review highlights the progressions, new methodologies, innovative instrumental modifications, and challenges for efficient protein analysis in human specimens, animal tissues, and plant samples. Protein analysis fields covered in this review include analysis of native, reduced, and denatured proteins in addition to Western blotting, protein therapeutics and proteomics.

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Developments in Interfacing Designs for CE–MS: Towards Enabling Tools for Proteomics and Metabolomics

Abstract: by representative examples in the fields of proteomics, glycomics and metabolomics. Finally, general conclusions and perspectives are provided.

Cited by 67 publications

References 77 publications

Label-free Quantification of Proteins in Single Embryonic Cells with Neural Fate in the Cleavage-Stage Frog (Xenopus laevis) Embryo using Capillary Electrophoresis Electrospray Ionization High-Resolution Mass Spectrometry (CE-ESI-HRMS)

Label-free Quantification of Proteins in Single Embryonic Cells with Neural Fate in the Cleavage-Stage Frog (Xenopus laevis) Embryo using Capillary Electrophoresis Electrospray Ionization High-Resolution Mass Spectrometry (CE-ESI-HRMS)

Analytical Pitfalls and Challenges in Clinical Metabolomics

Recent advances in protein analysis by capillary and microchip electrophoresis

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