Electrophoresis (ELECTROPHORESIS )

Publisher: Electrophoresis Society; International Electrophoresis Society, John Wiley and Sons


Attracting high-quality papers from the world's leading laboratories ELECTROPHORESIS has proved itself indispensable in the life sciences where electrophoresis is the most widely used method. ELECTROPHORESIS is the foremost journal for new analytical and preparative methods and for innovative applications on all aspects of electrophoresis. It has an international scope and publishes original papers short communications and review articles in the following areas: biochemistry molecular and cell biochemistry genetics immunology microbiology clinical chemistry forensics food science and many more. Special Issues Paper Symposium issues devoted to new developments prepared by an expert Guest Editor are published regularly with special emphasis on capillary electrophoresis (2-3 issues per year)and proteomics. The journal also publishes proceedings of international and national electrophoresis meetings.

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    Electrophoresis, Proteomics reviews
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John Wiley and Sons

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Publications in this journal

  • [Show abstract] [Hide abstract]
    ABSTRACT: To realise portable systems for routine measurements in point-of-care settings, microchip electrophoresis (ME) methods are required to be robust across many single-use chips. While it's well-known internal standards (ISTDs) improve run-to-run precision, a systematic investigation is necessary to determine the significance of chip-to-chip imprecision in ME and how ISTDs account for this. This paper addresses this question by exploring the reproducibility of Na quantification across six basic, in-house fabricated microchips. A dataset of 900 electrophoerograms was collected from analysing five concentrations of NaCl with two ISTDs (CsCl and LiCl). While both improved the peak area reproducibility, the Na/Cs ratio was superior to the Na/Li ratio (improving the RSD by a factor of 2 to 4, depending on the Na concentration). We attribute this to the significant variation in microchannel surface properties, which was accounted for by cesium but not lithium. Microchip dimension and detector variations were only a few percent, and could be improved through commercial fabrication over in-house made microchips. These results demonstrate that ISTDs not only correct for intra-chip imprecision, but are also a viable means to correct for chip-to-chip imprecision inherent in disposable, point-of-care ME devices. However, as expected, the internal standard must be carefully chosen. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Electrophoresis 12/2014;
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    ABSTRACT: For forensic and population genetic purposes, a total of 125 unrelated volunteers' blood samples were collected from Chinese Bai ethnic minority group to analyze sequence variation of two hypervariable segments HVS-I and HVS-II in the mitochondrial DNA control region. Comparing the HVS-I and HVS-II sequences of the 125 Chinese Bais to the Anderson reference sequence, we found 86 polymorphic loci in HVS-I and 40 in HVS-II in mitochondrial DNA sequences of the Chinese Bai ethnic minority group, which defined 93 and 53 different haplotypes, respectively. Haplotype diversity and the mean pairwise differences were 0.992 ± 0.003 and 6.553 in HVS-I, and 0.877 ± 0.027 and 2.407 in HVS-II, respectively. We defined 4 macrohaplogroups R, M, N and D with the proportions ranging from 9.6% to 40.0%. With the analysis of the hypervariable domain from nucleotide 16180 to 16193 in HVS-I, our study revealed new hyplotypes of sequence variations. In addition, the Fst metric, phylogenetic tree and principal component analysis demonstrated a close genetic relationship between the Bai group and Chinese Han populations from South China, Changsha and Guangdong. The results support that the Bai group is a multi-origin ethnic minority which has merged with the Chinese Han population. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Electrophoresis 12/2014;
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    ABSTRACT: Insulator-based dielectrophoresis (iDEP) employs insulating structures embedded in a microchannel to produce electric field gradients. This contribution presents a detailed analysis of the regions within an iDEP system where particles are likely to be retained due to dielectrophoretic trapping in a microchannel with an array of cylindrical insulating structures. The effects of particle size and shape on dielectrophoretic trapping were analyzed by employing 1 μm and 2 μm polystyrene particles and E. coli cells. This research aims to study the mechanism behind dielectrophoretic trapping and develop a deeper understanding of iDEP systems. Mathematical modeling with COMSOL Multiphysics was employed to assess electrokinetic and dielectrophoretic particle velocities. Experiments were carried out to determine the location of dielectrophoretic barriers that block particle motion within an iDEP microchannel; this supported the estimation of a correction factor to match experiments and simulations. Particle velocities were predicted with the model, demonstrating how the different forces acting on the particles are in equilibrium when particle trapping occurs. The results showed that particle size and shape have a significant effect on the magnitude, location and shape of the regions of dielectrophoretic trapping of particles, which are defined by DEP iso-velocity lines and EK iso-velocity lines. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Electrophoresis 12/2014;
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    ABSTRACT: Thymidine kinase-1 (TK1) is an important cancer biomarker whose serum levels are elevated in early cancer development. We developed a microchip electrophoresis immunoaffinity assay to measure recombinant purified TK1 (pTK1) using an antibody that binds to human TK1. We fabricated poly(methyl methacrylate) microfluidic devices to test the feasibility of detecting antibody (Ab)-pTK1 immune complexes as a step towards TK1 analysis in clinical serum samples. We were able to separate immune complexes from unbound antibodies using 0.5X phosphate buffer saline (pH 7.4) containing 0.01% Tween-20, with 1% w/v methylcellulose that acts as a dynamic surface coating and sieving matrix. Separation of the antibody and Ab-pTK1 complex was observed within a 5 mm effective separation length. This method of detecting pTK1 is easy to perform, requires only a 10 μL sample volume, and takes just 1 minute for separation. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Electrophoresis 12/2014;
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    ABSTRACT: The objective of this study was to characterize the differences in electrophoretic behavior between linear and branched PEG-conjugated proteins. Human growth hormone and alpha-lactalbumin modified by linear or branched PEGs with molecular weight of 10 kDa were analyzed by size-exclusion chromatography (SEC), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), SDS-PAGE, and microchip CGE (MCGE). Chromatographic and mass spectrometric differences between the linear and branched PEG-proteins on SEC and MALDI-TOF MS were small, but their electrophoretic behaviors on SDS-PAGE and MCGE were significantly different. In particular, MCGE showed significant differences in the peak width and the migration times of linear and branched PEG-proteins, in which the branched PEG-proteins exhibited a narrower peak and longer migration time than the linear PEG-proteins. This phenomenon may explain the longer circulation half-life for the branched PEG-proteins observed in previously reported in vivo studies. Consequently, this study indicates that MCGE may be a valuable tool for differentiating linear and branched PEG-proteins. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Electrophoresis 12/2014;
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    ABSTRACT: Pt-based anti-cancer drugs, such as cisplatin, are known to undergo several (bio-) chemical transformation steps after administration. Hydrolysis and adduct formation with small nucleophiles and larger proteins are their most relevant reactions on the way to the final reaction site (DNA), but there are still many open questions regarding the identity and pharmacological relevance of various proposed adducts and intermediates. Furthermore, the role of buffer components or additives, which are inevitably added to samples during any type of analytical measurement, has been frequently neglected in previous studies. Here, we report on adduct formation reactions of the fluorescent cisplatin analogue carboxyfluorescein diacetate-platinum (CFDA-Pt) in commonly used buffers and cell culture medium. Our results indicate that chelation reactions with non-innocent buffers (e.g. Tris) and components of the cell culture / cell lysis medium must be taken into account when interpreting results. Adduct formation kinetics was followed up to 60 hours at nM concentrations of CFDA-Pt by using CE-LIF. CE-MS enabled the on-line identification of such unexpected adducts down to the nanomolar concentration range. By using an optimized sample preparation strategy, unwanted adducts can be avoided and several fluorescent adducts of CFDA-Pt are detectable in sensitive and cisplatin-resistant cancer cell lines. By processing samples rapidly after incubation, we could even identify the initial, but transient, Pt-species in the cells as deacetylated CFDA-Pt with unaltered complexing environment at Pt. Overall, the proposed procedure enables a very sensitive and accurate analysis of low-molecular-mass Pt-species in cancer cells, involving a fast CE-LIF detection within five minutes. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Electrophoresis 12/2014;
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    ABSTRACT: In recent years, there are significant interests in the manipulation of bipolar Janus particles. In this article we investigate the transient behavior of the electro-orientation process and particle-particle interaction of ellipsoidal bipolar Janus particles in the presence and absence of a DC electric field. The bipolar particle dynamics is modeled with a body force term in the fluid flow equations based on the Maxwell stress tensor. This force is due to presence of bipolar surface charges on the particles as well as their interactions with an imposed field. An interface resolved numerical scheme that consider the finite size of the particle is adopted for computation of the electric and flow fields. Our numerical results show that in the absence of an electric field, particles can undergo self-orientation to reach an equilibrium position. The time taken to reach a stable orientation depends on the initial configuration and inter-particle separation distance. Bipolar particles experience forces only on their polar ends, a phenomena that is difficult to capture with non-interface resolved methods. When bipolar particles are exposed to an external electric field, they rotate to align along the external electric field direction. Depending upon the initial configuration, particles orient via clockwise or counter clockwise rotations to form head to tail chains. The time required to form particle assembly strongly depends on particle size and bipolar charge density. The present numerical algorithm can be applied to a wider class of dual-faced Janus particles. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Electrophoresis 12/2014;
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    ABSTRACT: Although numerous biomarkers or biomarker candidates have been discovered to detect levels of drinking and intervals of time after last drinking episode, only a few biomarkers have been applied to monitor abstinence in a longer interval (≥ 6 weeks) from alcohol abuse. Considering sample sources, sensitivity, and specificity, new biomarkers from blood with better accuracy are needed. To address this, serum proteomic profiles were compared between pre- and post- treatment samples from subjects seeking treatment for alcohol abuse and dependence in an intensive 6-week daily outpatient program using high-abundance plasma protein immunodepletion and LC-MS/MS techniques. Protein identification, quantification, candidate biomarker selection, and prioritization analyses were carried out. Among the 246 quantified serum proteins, abundance of 13 and 45 proteins in female and male subjects were significantly changed (p ≤ 0.05), respectively. Of these biomarker candidate proteins, 2 (female) and 8 (male) proteins were listed in category 1, with high area under the receiver operating characteristic (ROC) curve (AUC), sensitivity, specificity, and fold change. In summary, several new biomarker candidates have been identified to monitor abstinence from alcohol abuse. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Electrophoresis 12/2014;
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    ABSTRACT: For Gaussian peaks, the migration time of the analyte results as the position of the top of the peak and the zone variance is proportional to the peak width. Similar relations have not yet been derived for the Haarhoff-van der Linde (HVL) function, which appears as a fundamental peak shape function in electrophoresis. We derive the relations between the geometrical measures of the HVL-shaped peak, i.e. the position of its maximum, its width and a measure of its asymmetry, and the respective parameters, a1 , a2 and a3 , of the corresponding HVL function. Under the condition of the HVL-shaped peak, the a1 parameter reflects the true migration time of the analyte, which may differ from the peak top position significantly. Our procedure allows us to express the parameters without the need of any external data processing (nonlinear regression). We demonstrate our approach on simulated peaks and on experimental data integrated by the ChemStation software (delivered with the CE instrumentation by Agilent Technologies). A significant improvement is achieved reading the migration time of the experimental and simulated peaks, which draws the error of the HVL-shaped peak migration time evaluation down to the resolution of the data sampling rate. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Electrophoresis 12/2014;
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    ABSTRACT: In this manuscript, Langevin Dynamics simulations and Tension-Propagation theory are used to investigate the forced translocation of a polymer from a confining tube through a nanopore situated at one of the tube's ends. The diameter of the tube allows for a control over the polymer conformations: decreasing the tube diameter reduces the number of conformations available to the polymer chain both before and during translocation. As the tube diameter is decreased, the translocation time is observed to increase. Interestingly, while the width of the distribution of translocation times is reduced if the chain starts in a tube, it reaches a maximum for weakly confining tubes. A Tension-Propagation approach is developed for the tube-nanopore setup in the strongly-driven limit. Good agreement between the simulations and the theory allows for an exploration of the underlying physical mechanisms, including the calculation of an effective pore friction and the assessing of the impact of monomer crowding on the trans side. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Electrophoresis 12/2014;
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    ABSTRACT: To shed light on the multistep process of squamous cell carcinoma development and the underlying pathologic mechanisms, we performed comparative proteome analysis of keratinocytes, keratinocytes stimulated with Il-1beta and A431 epidermoid carcinoma cells. Fractionation of the cells into supernatant, nucleus and cytoplasm was followed by protein separation, proteolytic digest and nano-LC separation and fragmentation using an ion trap mass spectrometer. Specific bioinformatics tools were used to generate a list of keratinocyte-specific proteins. Ninety percent of these proteins were found to be upregulated in keratinocytes versus the A431 cells. Classification of the identified proteins by biologic function and GSEA revealed that keratinocytes produced more proteins involved in cell differentiation, cell adhesion, cell junction, calcium ion, calmodulin binding, cytoskeleton organization and cytokinesis, whereas A431 produced more proteins involved in cell cycle checkpoint, cell cycle process, RNA processing and transport, DNA damage and repair, RNA and DNA binding and chromatin remodeling. The protein signatures of A431 and normal keratinocytes treated with IL-1beta showed marked similarity, confirming that inflammation is an important step in malignant transformation in NMSC. Thus, proteome profiling and bioinformatic processing may support the understanding of the underlying mechanisms, with the potential to facilitate development of early biomarkers and patient-tailored therapy. This article is protected by copyright. All rights reserved.
    Electrophoresis 11/2014;
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    ABSTRACT: In this work, the suitability of a methodology based on dispersive liquid-liquid microextraction (DLLME) has been evaluated for the extraction of four endoestrogens (estriol, 17α-estradiol, 17β-estradiol and estrone), an exoestrogen (17α-etynylestradiol) and a mycotoxin (zearalenone), together with some of their major metabolites (2-methoxyestradiol, α-zearalanol, β-zearalanol, α-zearalenol and β-zearalenol) from different types of milk (whole and skimmed cow milk and semi-skimmed goat milk) and whole natural yogurt. The methodology includes a previous protein precipitation with acidified acetonitrile and a defatting step with n-hexane. Separation of the analytes, determination and quantification were developed by micellar electrokinetic chromatography (MEKC) coupled to electrospray ionization mass spectrometry (ESI-MS) using a background electrolyte containing an aqueous solution of ammonium perfluorooctanoate as MS friendly surfactant. Calibration, precision and accuracy studies of the described DLLME-MEKC-MS/MS method were evaluated obtaining a good linearity and limits of detection in the low μg/L range. This article is protected by copyright. All rights reserved.
    Electrophoresis 11/2014;
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    ABSTRACT: Inflammatory processes and other failures related to the immune system are common features associated with Alzheimer's disease, in both brain and the peripheral system. Thus, the study of the main organs of the immune system may have a great potential for the elucidation of pathological mechanisms underlying these abnormalities. This is the first metabolomic investigation performed in spleen and thymus from transgenic mice of Alzheimer's disease. Tissues were fingerprinted using a metabolomic platform comprising gas chromatography-mass spectrometry and ultra-high performance liquid chromatography-mass spectrometry. Multivariate statistics demonstrated significant differences in numerous metabolites between the APP/PS1 mice and wild type controls, and it was proven that multiple biochemical pathways are disturbed in these organs including abnormal metabolism of phospholipids, energy deficiencies, altered homeostasis of amino acids, oxidative stress, and others. Therefore, these findings highlight the importance of the proper metabolic functioning of peripheral immune system in the development of neurodegenerative disorders such as Alzheimer's disease. This article is protected by copyright. All rights reserved.
    Electrophoresis 11/2014;
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    Electrophoresis 08/2014; 35(15):2067.
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    ABSTRACT: A new MCE method for the determination of oxalic, citric, glycolic, lactic, and 2‐ and 3‐hydroxybutyric acids, indicators of some metabolic and neurological diseases, in cerebrospinal fluid (CSF) was developed. MCE separations were performed on a PMMA microchip with coupled channels at lower pH (5.5) to prevent proteins interference. A double charged counter‐ion, BIS‐TRIS propane, was very effective in resolving the studied organic acids. The limits of detection (S/N = 3) ranging from 0.1 to 1.6 μM were obtained with the aid of contact conductivity detector implemented directly on the microchip. RSDs for migration time and peak area of organic acids in artificial and CSF samples were Keywords: Cerebrospinal fluid; MCE; Micro‐SPE; Organic acids Document Type: Research Article DOI: http://dx.doi.org/10.1002/elps.201300455 Publication date: August 1, 2014 $(document).ready(function() { var shortdescription = $(".originaldescription").text().replace(/\\&/g, '&').replace(/\\, '<').replace(/\\>/g, '>').replace(/\\t/g, ' ').replace(/\\n/g, ''); if (shortdescription.length > 350){ shortdescription = "" + shortdescription.substring(0,250) + "... more"; } $(".descriptionitem").prepend(shortdescription); $(".shortdescription a").click(function() { $(".shortdescription").hide(); $(".originaldescription").slideDown(); return false; }); }); Related content In this: publication By this: publisher By this author: Danč, Ladislav ; Bodor, Róbert ; Troška, Peter ; Horčičiak, Michal ; Masár, Marián GA_googleFillSlot("Horizontal_banner_bottom");
    Electrophoresis 08/2014; 35(15).
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    ABSTRACT: Human nail is an important forensic material for parental testing and individual identification in large-scale disasters. Detection of short tandem repeat (STR) polymorphism from hard tissues generally requires DNA purification, which is technically complicated and time consuming. In the present study, we attempted to detect STR polymorphisms from untreated human nail samples by direct polymerase chain reaction (PCR) amplification method using the primer mixture supplied with the GenePrint® SilverSTR® III System or the AmpFℓSTR® Identifiler® PCR Amplification Kit, and Tks Gflex DNA polymerase known to be effective for amplification from crude samples. A nail fragment measuring approximately 1.5 mm in breadth and 0.5 mm in length was placed directly into a PCR tube, and various PCR conditions were tested. The PCR products were analyzed by denaturing acrylamide gel electrophoresis or capillary electrophoresis. Multiple STR polymorphisms were detected successfully. This method that detects STR polymorphisms not only from fresh human fingernails, but also from old nail fragments stored at room temperature for up to 10 years is expected to become a novel DNA analytical method in forensic medicine and genetic studies. This article is protected by copyright. All rights reserved.
    Electrophoresis 06/2014;