Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips (2012–2014)

Breadmore, Michael C.; Tubaon, Ria Marni; Shallan, Aliaa I.; Phung, Sui Ching; Keyon, Aemi Syazwani Abdul; Gstoettenmayr, Daniel; Prapatpong, Pornpan; Alhusban, Ala A.; Ranjbar, Leila; See, Hong Heng; Dawod, Mohamed; Quirino, Joselito P.

doi:10.1002/elps.201400420

Cited by 142 publications

(105 citation statements)

References 247 publications

(267 reference statements)

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“…To assess proteins at trace levels, various CE approaches can help enrich molecules on-column and separate them in increased peak capacity (64). New-generation interfaces that minimize/ eliminate sample dilution between CE and ESI may be used to ionize peptides more efficiently (see reviews in Ref (33,(65)(66)(67)); electrokinetically pumped sheath-flow (32,38) and sheathless (68,69) interfaces are promising designs in this direction. Based on recent successes in proteome coverage and post-translational analysis by CE (35,37,69), we expect these innovative solutions combined with new-generation FIG.…”

Section: Label-free Quantification Of Single Embryonic Cellsmentioning

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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Section: Label-free Quantification Of Single Embryonic Cellsmentioning

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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“…Although recent reviews by El Deeb et al [66] and Breadmore et al [67] show that extensive efforts have been made to improve the sensitivity of CE, CE-MS remains mostly applied in proteomics, metabolomics and enzymatic studies. The quantification of pharmaceutical compounds is less common; however, due to its selectivity CE-MS can be used to differentiate and quantify enantiomers.…”

Section: (I) State-of-the-art Chromatographymentioning

confidence: 99%

Quantitative mass spectrometry methods for pharmaceutical analysis

Loos¹,

Schepdael²,

Cabooter³

2016

Phil. Trans. R. Soc. A.

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Quantitative pharmaceutical analysis is nowadays frequently executed using mass spectrometry. Electrospray ionization coupled to a (hybrid) triple quadrupole mass spectrometer is generally used in combination with solid-phase extraction and liquid chromatography. Furthermore, isotopically labelled standards are often used to correct for ion suppression. The challenges in producing sensitive but reliable quantitative data depend on the instrumentation, sample preparation and hyphenated techniques. In this contribution, different approaches to enhance the ionization efficiencies using modified source geometries and improved ion guidance are provided. Furthermore, possibilities to minimize, assess and correct for matrix interferences caused by co-eluting substances are described. With the focus on pharmaceuticals in the environment and bioanalysis, different separation techniques, trends in liquid chromatography and sample preparation methods to minimize matrix effects and increase sensitivity are discussed. Although highly sensitive methods are generally aimed for to provide automated multi-residue analysis, (less sensitive) miniaturized set-ups have a great potential due to their ability for in-field usage. This article is part of the themed issue ‘Quantitative mass spectrometry’.

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“…This boundary can be created by changes in conductivity (as in field-amplified sample stacking and isotachophoresis), or changes in pH (as in dynamic pH junction), or inclusion of a pseudostationary phase (as in sweeping and micelle collapse) [22][23][24][25]. A series of biannual reviews, first published in Electrophoresis in 2007 [26] and continued in 2009 [27], 2011 [28], 2013 [29], and 2015 [30] To the best of our knowledge, some reviews and book chapters on IL-based surfactants cover the use of ILs in CE [32,33], but in the present work, for the first time, we described the applications resulted from implementing ILs in enhancing the sensitivity of CE. Figure 2 shows a chart summarizing the different modes based on ILs in enhancing the sensitivity of CE.…”

Section: Introductionmentioning

confidence: 99%

Ionic liquids in enhancing the sensitivity of capillary electrophoresis: Off‐line and on‐line sample preconcentration techniques

2016

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The popularity of ionic liquids (ILs) has grown during the last decade in enhancing the sensitivity of CE through different off-line or on-line sample preconcentration techniques. Water-insoluble ILs were commonly used in IL-based liquid phase microextraction, in all its variants, as off-line sample preconcentration techniques combined with CE. Water-soluble ILs were rarely used in IL-based aqueous two phase system (IL-ATPS) as an off-line sample preconcentration approach combined with CE in spite of IL-ATPS predicted features such as more compatibility with CE sample injection due to its relatively low viscosity and more compatibility with CE running buffers avoid, in some cases, anion exchange precipitation. Therefore, the attentions for the key parameters affecting the performance of IL-ATPSs were generally presented and discussed. On-line CE preconcentration techniques containing IL-based surfactants at nonmicellar or micellar concentrations have become another interesting area to improve CE sensitivity and it is likely to remain a focus of the field in the endeavor because of their numerous to create rapid, simple and sensitive systems. In this article, significant contributions of ILs in enhancing the sensitivity of CE are described, and a specific overview of the relevant examples of their applications is also given.

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Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips (2012–2014)

Cited by 142 publications

References 247 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)

Quantitative mass spectrometry methods for pharmaceutical analysis

Ionic liquids in enhancing the sensitivity of capillary electrophoresis: Off‐line and on‐line sample preconcentration techniques

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