Recent applications of on-line sample preconcentration techniques in capillary electrophoresis

Kitagawa, Fumihiko; Otsuka, Koji

doi:10.1016/j.chroma.2013.10.066

Cited by 176 publications

(93 citation statements)

References 212 publications

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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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“…While this flow-induced particle accumulation has potentially adverse effects, such as in the clogging of filtration membranes, this accumulation can also be exploited for practical applications. For example, because this effect can locally concentrate colloidal particles almost to their maximum packing limit, it could be used for preconcentrating or separating particles in a short period of time for various analytical purposes [21,27].…”

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confidence: 99%

Accumulation of Colloidal Particles in Flow Junctions Induced by Fluid Flow and Diffusiophoresis

et al. 2017

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The flow of solutions containing solutes and colloidal particles in porous media is widely found in systems including underground aquifers, hydraulic fractures, estuarine or coastal habitats, water filtration systems, etc. In such systems, solute gradients occur when there is a local change in the solute concentration. While the effects of solute gradients have been found to be important for many applications, we observe an unexpected colloidal behavior in porous media driven by the combination of solute gradients and the fluid flow. When two flows with different solute concentrations are in contact near a junction, a sharp solute gradient is formed at the interface, which may allow strong diffusiophoresis of the particles directed against the flow. Consequently, the particles accumulate near the pore entrance, rapidly approaching the packing limit. These colloidal dynamics have important implications for the clogging of a porous medium, where particles that are orders of magnitude smaller than the pore width can accumulate and block the pores within a short period of time. We also show that this effect can be exploited as a useful tool for preconcentrating biomolecules for rapid bioassays.

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“…However, as injecting a large volume of sample will reduce the capillary length available for the separation, such injection will need to be followed by an in-capillary sample concentration step. Several stacking approaches [30,31], based on hydrodynamic or electrokinetic injection should be considered, but one has first to ensure that no disruption of the non-covalent aggregates occurs due to the additional physico-chemical stress applied to the sample during such stacking steps. Performing an immuno-extraction to concentrate the sample before its injection into the separation capillary [32,33] could be another way to analyze a larger volume of sample and therefore to improve the repeatability.…”

Section: Discussionmentioning

confidence: 99%

Advances and Pitfalls in the Capillary Electrophoresis Analysis of Aggregates of Beta Amyloid Peptides

Denoroy

Parrot

2017

Separations

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Alzheimer's disease is characterized by the accumulation of brain amyloid plaques composed of aggregates of amyloid β (Aβ) peptides. The present paper describes a novel and easy-to-run capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) method for the specific analysis of fibrillar forms of Aβ aggregates obtained after in vitro incubation of Aβ 1-40 monomer. For that purpose, an affinity CE-LIF approach in which the ligand thioflavine T was added to the running buffer has been used, leading to the separation and detection of various fibrillar aggregates which migrated as spikes. The procedure has been optimized to get spikes only corresponding to Aβ aggregates, through the careful elimination of interfering factors and the electrophoretic validation of the link between spikes and particulate material. This method exhibited semi-quantification capabilities, led to the separation of Aβ fibrillar aggregates of different sizes and showed that highly concentrated solutions of Aβ peptides led to the formation of aggregates of larger size than lower-concentrated solution did. Advances brought by this method as well as future development needed to overcome its present limitations are discussed.

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Recent applications of on-line sample preconcentration techniques in capillary electrophoresis

Cited by 176 publications

References 212 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)

Accumulation of Colloidal Particles in Flow Junctions Induced by Fluid Flow and Diffusiophoresis

Advances and Pitfalls in the Capillary Electrophoresis Analysis of Aggregates of Beta Amyloid Peptides

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