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Uroš Andjelković, Srdjan Tufegdžić, Milica Popović
The exclusive properties of monolithic supports enable fast mass transfer, high porosity, low back pressure, easy preparation process and miniaturisation and the availability of different chemistries make them particularly suitable materials for high-throughput (HTP) protein and peptide separation. In this review recent advances in monolith-based chromatographic supports for HTP screening of protein and peptide samples are presented and their application in HTP sample preparation (separation, enrichment, depletion, proteolytic digestion) for HTP proteomics is discussed...
September 14, 2017: Electrophoresis
Oier Aizpurua-Olaizola, Javier Sastre Torano, Aliaksei Pukin, Ou Fu, Geert Jan Boons, Gerhardus J de Jong, Roland J Pieters
Developing tools for the study of protein carbohydrate interactions is an important goal in glycobiology. Cholera toxin inhibition is an interesting target in this context, as its inhibition may help the fight against cholera. For the study of novel ligands an affinity capillary electrophoresis (ACE) method was optimised and applied. The method uses unlabeled cholera toxin B-subunit (CTB) and unlabeled carbohydrate ligands based on ganglioside GM1-oligosaccharides (GM1os). In an optimized method at pH 4, adsorption of the protein to the capillary walls was prevented by a polybrene-dextran sulfate-polybrene (PB-DS-PB) coating...
September 14, 2017: Electrophoresis
Huifen Ma, Yun Bai, Jin Li, Yan-Xu Chang
Bioactive ingredients of natural products can protect human body from harm, as well as prevent and treat disease. Screening bioactive compounds from natural products is attracting particular attention. It is a great challenge to separate and detect the active compounds from complex matrix natural products. Capillary electrophoresis (CE) plays a vital role in screening active compounds because of its unique features such as high-efficiency separation, short analysis time, minimal sample consumption and ease to realize automatization etc...
September 11, 2017: Electrophoresis
Deying Kong, Zilin Chen
Pillar[n]arenes have achieved much interest in material chemistry and supramolecular chemistry due to unusual pillar shape structure and high selectivity toward guest. However, pillar[n]arenes have not yet been applied in capillary electrochromatography. This work at first time reports that carboxylatopillar[5]arene is used as a stationary phase in open-tubular capillary electrochromatography. Carboxylatopillar[5]arene not only possess the advantages of pillar[n]arenes but also provide free carboxy groups for immobilizing on the inner wall of capillary column via covalent bonding...
September 11, 2017: Electrophoresis
B Moritz, V Locatelli, Michele Niess, Andrea Bathke, Steffen Kiessig, Barbara Entler, C Finkler, H Wegele, Jan Stracke
Capillary zone electrophoresis is a well-established technique for charge heterogeneity testing of biopharmaceuticals 1. It is based on the differences between the ratios of net charge and hydrodynamic radius. In an extensive intercompany study, it was recently shown that capillary zone electrophoresis is very robust and can be easily implemented in labs that did not perform it before 2. However, individual characteristics of some examined proteins resulted in suboptimal resolution. Therefore, enhanced method development principles where applied here to investigate possibilities for further method optimization...
September 9, 2017: Electrophoresis
Mariia А Stepanova, Lilia R Kinziabulatova, Anna A Nikitina, Evgenia G Korzhikova-Vlakh, Tatiana B Tennikova
The development of sorbents for selective binding of cholesterol, which is a risk factor for cardiovascular disease, has a great importance for analytical science and medicine. In this work, two series of macroporous cholesterol-imprinted monolithic sorbents differing in the composition of functional monomers (methacrylic acid, butyl methacrylate, 2-hydroxyethyl methacrylate and ethylene dimethacrylate), amount of a template (4, 6 and 8 mol%) used for molecular imprinting, as well as mean pore size were synthesized by in situ free-radical process in stainless steel housing of 50 mm × 4...
September 7, 2017: Electrophoresis
Federico J V Gomez, Adrian Spisso, María Fernanda Silva
A novel methodology is presented for the enhanced electrochemical detection of oleuropein in complex plant matrices by Graphene Oxide Pencil Grahite Electrode (GOPGE) in combination with a buffer modified with a Natural Deep Eutectic Solvent, containing 10% (v/v) of Lactic acid, Glucose and H2 O (LGH). The electrochemical behavior of oleuropein in the modified-working buffer was examined using differential pulse voltammetry. The combination of both modifications, NADES modified buffer and nanomaterial modified electrode, LGH-GOPGE, resulted on a signal enhancement of 5...
September 7, 2017: Electrophoresis
Liying Xu, Ruimiao Chang, Meng Chen, Lou Li, Yayun Huang, Hongfen Zhang, Anjia Chen
The purpose of this study was to develop a comprehensive, rapid and practical capillary electrophoresis (CE) method for quality control (QC) of Guan-Xin-Ning (GXN) injection based on fingerprint analysis and simultaneous separation and determination of seven constituents. In fingerprint analysis, a capillary zone electrophoresis (CZE) method with a running buffer of 30 mM borate solution (pH 9.3) was established. Meanwhile, 10 batches of samples were used to establish the fingerprint electropherogram and 34 common peaks were obtained within 20 min...
September 7, 2017: Electrophoresis
Simone Stratz, Pascal Emilio Verboket, Karina Hasler, Petra Stephanie Dittrich
Here, we present a multifunctional microfluidic device whose integrative design enables to combine cell culture studies and quantitative single cell biomolecule analysis. The platform consists of 32 analysis units providing two key features; first, a micrometer-sized trap for hydrodynamic capture of a single Saccharomyces cerevisiae (S. cerevisiae) yeast cell; second, a convenient double-valve configuration surrounding the trap. Actuating of the outer valve with integrated opening results in a partial isolation in a volume of 11...
September 7, 2017: Electrophoresis
Ales Podgornik, Masataka Hamachi, Yu Isakari, Noriko Yoshimoto, Shuichi Yamamoto
Effect of pore size on the performance of ion-exchange monolith tube chromatography of large biomolecules was investigated. Radial flow 1 mL polymer based monolith tubes of different pore sizes (1.5, 2 and 6 μm) were tested with model samples such as 20mer poly T DNA, basic proteins and acidic proteins (molecular weight 14000-670000). Pressure drop, pH transient, the number of binding site, dynamic binding capacity and peak width were examined. Pressure drop-flow rate curves and dynamic binding capacity values were well correlated with the nominal pore size...
September 5, 2017: Electrophoresis
Weiyu Liu, Yukun Ren, Ye Tao, Bobin Yao, You Li
We report herein field-effect control on in-phase electrothermal streaming from a theoretical point of view, a phenomenon termed "AC electrothermal-flow field effect transistor" (ACET-FFET), in the context of a new technology for handing analytes in microfluidics. Field-effect control through a gate terminal endows ACET-FFET the ability to generate arbitrary symmetry breaking in the transverse vortex flow pattern, which makes it attractive for mixing microfluidic samples. A computational model is developed to study the feasibility of this new microfluidic device design for micromixing...
September 5, 2017: Electrophoresis
Nour Noaman, Prabhodh S Abbineni, Michael Withers, Jens R Coorssen
Modified colloidal Coomassie Brilliant Blue (cCBB) staining utilising a novel destain protocol and near-infrared fluorescence detection (nIRFD) rivals the in-gel protein detection sensitivity (DS) of SYPRO Ruby. However, established DS estimates are likely inaccurate in terms of 2DE-resolved proteoform 'spots' since DS is routinely measured from comparatively diffuse protein 'bands' following wide-well 1DE. Here, cCBB DS for 2DE-based proteomics was more accurately determined using narrow-well 1DE. As precise estimates of protein standard monomer concentrations are essential for accurate quantitation, coupling UV absorbance with gel-based purity assessments is described...
September 5, 2017: Electrophoresis
Xuan Wang, Nengsheng Ye
In the field of analytical chemistry, sample preparation and chromatographic separation are two core procedures. The means by which to improve the sensitivity, selectivity and detection limit of a method have become a topic of great interest. Recently, porous organic frameworks, such as metal-organic frameworks (MOFs) and covalent organic frameworks (COFs), have been widely used in this research area because of their special features, and different methods have been developed. This review summarizes the applications of MOFs and COFs in sample preparation and chromatographic stationary phases...
September 4, 2017: Electrophoresis
Meiling Wen, Ya Jin, Takashi Manabe, Shumin Chen, Wen Tan
MS identification has long been used for PAGE-separated protein bands, but global and systematic quantitation utilizing MS after PAGE has remained rare and not been reported for native PAGE. Here we reported on a new method combining native PAGE, whole-gel slicing and quantitative LC-MS/MS, aiming at comparative analysis on not only abundance, but also structures and interactions of proteins. A pair of human plasma and serum samples were used as test samples and separated on a native PAGE gel. Six lanes of each sample were cut, each lane was further sliced into thirty-five 1...
September 4, 2017: Electrophoresis
Wenjing Zhang, Muyi He, Tao Yuan, Wei Xu
The measurement of electroosmotic flow (EOF) is important in a capillary electrophoresis (CE) experiment in terms of performance optimization and stability improvement. Although several methods exist, there are demanding needs to accurately characterize ultra-low electroosmotic flow rates (EOF rates), such as in coated capillaries used in protein separations. In this work, a new method, called the two-step method, was developed to accurately and rapidly measure EOF rates in a capillary, especially for measuring the ultra-low EOF rates in coated capillaries...
September 4, 2017: Electrophoresis
Daniel A Ribeiro, Douglas F Passos, Helen C Ferraz, Leda R Castilho
This work investigated the use of hydrophobic interaction membrane chromatography for intermediate purification of recombinant human Factor IX (rFIX) produced by CHO cells. The first purification step was based on a strong anion exchange monolith, thus forming a purification process fully based on convective media, which allow operation at high flow rates and low pressure drops, as well as modular scale-up. Although the starting material was challenging (CHO cell culture supernatant harvested at 70% cell viability), the two-step purification process showed promising results, with a global purification factor of 298, a global recovery of 69%, and DNA and endotoxin levels close to regulatory limits...
September 4, 2017: Electrophoresis
Jana Srbova, Pavla Krulisova, Lucie Holubova, Iago Pereiro, Amel Bendali, Audrey Hamiot, Veronika Podzemna, Jan Macak, Bruno Dupuy, Stephanie Descroix, Jean-Louis Viovy, Zuzana Bilkova
The success of microfluidic immunocapture based on magnetic beads depends primarily on a sophisticated microscale separation system and on the quality of the magnetic immunosorbent. A microfluidic chip containing a magnetically stabilized fluidized bed (μMSFB), developed for the capture and on-chip amplification of bacteria, was recently described by Pereiro et al. (2017). The present work shows the through development of an anti-Salmonella magnetic immunosorbents with the optimal capture efficiency and selectivity...
September 4, 2017: Electrophoresis
Margreet van den Berge, Titia Sijen
Messenger RNA (mRNA) profiling is a technique increasingly applied for the forensic identification of body fluids and skin. More recently, an mRNA-based organ typing assay was developed which allows for the inference of brain, lung, liver, skeletal muscle, heart, kidney and skin tissue. When applying this organ typing system in forensic casework the presence of animal, rather than human, tissue is an alternative scenario to be proposed, for instance that bullets carry cell material from a hunting event. Even though mRNA profiling systems are commonly in silico designed to be primate-specific, physical testing against other animal species is generally limited...
September 1, 2017: Electrophoresis
Veronica Mantovani, Fabio Galeotti, Francesca Maccari, Nicola Volpi
This article illustrates the basis and applications of methodologies for the analysis of simple and complex carbohydrates by means of capillary electrophoresis (CE). After a description of the most common and novel approaches useful for the analysis and characterization of carbohydrates, this review covers the recent advances in CE separation of monosaccharides, oligosaccharides and polysaccharides. Various CE techniques are also illustrated for the study of carbohydrates derived from complex glyco-derivatives such as glycoproteins and glycolipids, essential for biopharmaceutical and glycoproteomics applications as well as for biomarker detection...
August 30, 2017: Electrophoresis
Terry M Phillips
CE and microchip CE (ME)are powerful tools for the analysis of a number of different analytes and have been applied to a variety of clinical fields and human samples. This review will present an overview of the most recent applications of these techniques to different areas of clinical medicine during the period of 2014 to mid-2017. CE and ME have been applied to clinical chemistry, drug detection and monitoring, hematology, infectious diseases, oncology, endocrinology, neonatology, nephrology, and genetic screening...
August 29, 2017: Electrophoresis
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