The Experts below are selected from a list of 318 Experts worldwide ranked by ideXlab platform
Xavier Bossuyt - One of the best experts on this subject based on the ideXlab platform.
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interferences in clinical Capillary Zone Electrophoresis of serum proteins
Electrophoresis, 2004Co-Authors: Xavier BossuytAbstract:Capillary Zone Electrophoresis (CZE) of serum proteins has become a well-accepted method for the separation of serum proteins and for the detection of monoclonal proteins in clinical laboratories. As CZE uses ultraviolet detection for direct protein quantification, exogenous ultraviolet-absorbing substances are a novel challenge, not present in conventional gel-based methods in which protein is quantified by dye binding. This minireview gives a survey of interfering substances, including iodinated contrast agents and antibiotics.
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separation of serum proteins by automated Capillary Zone Electrophoresis
Clinical Chemistry and Laboratory Medicine, 2003Co-Authors: Xavier BossuytAbstract:Capillary Zone Electrophoresis (CZE) of serum proteins is increasingly gaining impact in clinical laboratories. Two automated multichannel instruments dedicated to the separation of serum proteins have become available over the last 6 years, the Paragon CZE 2000 (Beckman Coulter, CA, USA) and, more recently, the Capillarys (Sebia, France). This review focuses on the performance of these commercial instruments to separate serum proteins in a clinical laboratory setting. The utility of CZE to recognize various dysproteinemias and to detect and identify monoclonal proteins will be described and systematically reviewed. The reader will be provided with a summation and an understanding of CZE-specific interference.
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interference of radio opaque agents in clinical Capillary Zone Electrophoresis
Clinical Chemistry, 1999Co-Authors: Xavier Bossuyt, Alex Mewis, Norbert BlanckaertAbstract:Capillary Zone Electrophoresis (CZE) has emerged as a novel technique for the rapid and effective separation of serum proteins (1)(2)(3)(4)(5)(6)(7)(8). Recently, a multichannel automated system for CZE of human serum proteins (Paragon 2000 clinical Capillary Electrophoresis system; Beckman Instruments) became commercially available; this system offers a clinically reliable alternative to cellulose acetate and agarose Electrophoresis. CZE has the advantage of automation, improved precision, and a faster turnaround time (6)(8). In the conventional methods, quantification of the protein fractions is based on dye binding, whereas CZE uses ultraviolet detection at 214 nm for direct protein quantification via the peptide bonds. We asked whether intravascular agents such as radio-contrast media that absorb at 214 nm would simulate a monoclonal component on CZE. We performed high resolution agarose Electrophoresis with the Hydrasys analyzer (Sebia), using Hydragel 15 HR gels (Sebia) according to the …
Wolfgang Thormann - One of the best experts on this subject based on the ideXlab platform.
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Analysis of genetic variants of transferrin in human serum after desialylation by Capillary Zone Electrophoresis and Capillary isoelectric focusing.
IEEE Journal of Solid-state Circuits, 2017Co-Authors: Jitka Caslavska, Christian Lanz, Patricie Burda, Micha Tobler, Wolfgang ThormannAbstract:Capillary Electrophoresis analysis of transferrin in human serum is used to assess genetic variants after desialylation with neuraminidase and iron saturation to reduce the complexity of the transferrin pattern and thus facilitate the recognition of transferrin polymorphisms. Asialo-transferrin forms are analyzed by Capillary Zone Electrophoresis using assay conditions as for the monitoring of carbohydrate-deficient transferrin or by Capillary isoelectric focusing in a pH 5-8 gradient which requires immunoextraction of transferrin prior to analysis. With the carrier ampholytes used, peaks for iron saturated and iron depleted transferrin are monitored which indicates complexation of iron ions by carrier ampholytes. For BC, CD, and BD genetic variants, the expected peaks for B, C, and D forms of transferrin were detected with both methods. Monitoring of CC patterns revealed three cases, namely those producing double peaks in both methods, a double peak in Capillary isoelectric focusing only and a double peak in Capillary Zone Electrophoresis only. For all samples analyzed, data obtained by Capillary isoelectric focusing could be confirmed with gel isoelectric focusing. The two Capillary Electrophoresis methods are shown to represent effective tools to assess unusual transferrin patterns, including genetic variants with dissimilar abundances of the two forms.
Salvatore Fanali - One of the best experts on this subject based on the ideXlab platform.
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use of charged and neutral cyclodextrins in Capillary Zone Electrophoresis enantiomeric resolution of some 2 hydroxy acids
Journal of Chromatography A, 1993Co-Authors: Annalisa Nardi, Petr Boček, Alexey Eliseev, Salvatore FanaliAbstract:Abstract Enantiomers of racemic 2-hydroxy acids, namely 2-phenyllactic, 3-phenyllactic, mandelic, m-hydroxymandefic, p-hydroxymandelic and 3,4-hydroxymandelic acid, were resolved by Capillary Zone Electrophoresis. The separation was achieved by using the background electrolyte with addition of cyclodextrins. The effects of the type of cyclodextrin, the pH of the electrolyte and the shape of analyte compounds on the migration time and resolution were studied. Good resolution was obtained with background electrolytes in the pH range 5–7, supplemented with 2-hydroxypropyl, 6A-methylamino- and 6A 6D-dimethylamino- β-cyclodextrin.
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some separations of black and red water soluble fiber tip pen inks by Capillary Zone Electrophoresis and thin layer chromatography
Journal of Forensic Sciences, 1991Co-Authors: Salvatore Fanali, Marti SchudelAbstract:Capillary Zone Electrophoresis was tried for analyzing water-soluble fiber-tip pen inks from documents. The separation process for the compounds—most of them move as anions—is usually less than 10 min. Some examples of possible applications, also combined with thin-layer chromatography, are given.
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use of cyclodextrins in Capillary Zone Electrophoresis resolution of terbutaline and propranolol enantiomers
Journal of Chromatography A, 1991Co-Authors: Salvatore FanaliAbstract:Abstract Terbutaline and propranolol were resolved using Capillary Zone Electrophoresis. The effect of the type and the amount of cyclodextrins added to the background electrolyte on the migration time and the resolution of their enantiomers was studied. Good resolution of the racemic terbutaline was obtained using phosphate buffer at pH 2.5 containing either 5 m M heptakis(2,6-di-O-methyl)-β-cyclodextrin or 15 m M β-cyclodextrin. The background electrolyte, 50 m M phosphate buffer (pH—2.5) − 4 M urea − 40 m M β-cyclodextrin in 30% (v/v) methanol, on the other hand, gave the best resolution of propranolol enantiomers.
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Capillary Zone Electrophoresis and mass spectrometry for the characterization of genetic variants of human hemoglobin
Analytical Biochemistry, 1991Co-Authors: Pasquale Ferranti, Salvatore Fanali, Annalisa Nardi, Antonio Malorni, Pietro Pucci, L. OssiciniAbstract:Abstract This paper describes a simple and rapid analytical method for the structural identification of abnormal human hemoglobins. Globin chains obtained by precipitation of erythrocyte hemolysate in cold acetone are directly analyzed by Capillary Zone Electrophoresis in coated capillaries without any prior treatment. The speed and the high resolving power of Capillary Zone Electrophoresis allow fast differentiation of hemoglobins with similar charges. Capillary Zone electrophoretic tryptic mapping has also been performed for each globin, so that complete variant characterization can be achieved by direct comparison of the variant tryptic map with the corresponding normal one. Coupling electrophoretic data with analysis of enzymatic digests by mass spectrometry according to the “fast atom bombardment mapping” procedure makes it possible to quickly identify amino acid variations. This paper describes how the method can be applied to the characterization of common and uncommon variants and underlines the advantages and limitations of the procedure along with its potential uses in structural analysis of proteins.
Jitka Caslavska - One of the best experts on this subject based on the ideXlab platform.
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Analysis of genetic variants of transferrin in human serum after desialylation by Capillary Zone Electrophoresis and Capillary isoelectric focusing.
IEEE Journal of Solid-state Circuits, 2017Co-Authors: Jitka Caslavska, Christian Lanz, Patricie Burda, Micha Tobler, Wolfgang ThormannAbstract:Capillary Electrophoresis analysis of transferrin in human serum is used to assess genetic variants after desialylation with neuraminidase and iron saturation to reduce the complexity of the transferrin pattern and thus facilitate the recognition of transferrin polymorphisms. Asialo-transferrin forms are analyzed by Capillary Zone Electrophoresis using assay conditions as for the monitoring of carbohydrate-deficient transferrin or by Capillary isoelectric focusing in a pH 5-8 gradient which requires immunoextraction of transferrin prior to analysis. With the carrier ampholytes used, peaks for iron saturated and iron depleted transferrin are monitored which indicates complexation of iron ions by carrier ampholytes. For BC, CD, and BD genetic variants, the expected peaks for B, C, and D forms of transferrin were detected with both methods. Monitoring of CC patterns revealed three cases, namely those producing double peaks in both methods, a double peak in Capillary isoelectric focusing only and a double peak in Capillary Zone Electrophoresis only. For all samples analyzed, data obtained by Capillary isoelectric focusing could be confirmed with gel isoelectric focusing. The two Capillary Electrophoresis methods are shown to represent effective tools to assess unusual transferrin patterns, including genetic variants with dissimilar abundances of the two forms.
Thormann Wolfgang - One of the best experts on this subject based on the ideXlab platform.
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High-resolution Capillary Zone Electrophoresis and mass spectrometry for distinction of undersialylated and hypoglycosylated transferrin glycoforms in body fluids.
'Wiley', 2020Co-Authors: Caslavsky Jitka, Schild Christof, Thormann WolfgangAbstract:High-resolution Capillary Zone Electrophoresis is used to distinguish transferrin glycoforms present in human serum, cerebrospinal fluid, and serum treated with neuraminidase and N-glycosidase F. The obtained data are compared to mass spectrometry data from the literature. The main focus is on the analysis of the various asialo-transferrin, monosialo-transferrin, and disialo-transferrin molecules found in these samples. The features of Capillary Zone Electrophoresis and mass spectrometry are reviewed and highlighted in the context of the analysis of undersialylated and hypoglycosylated transferrin molecules. High-resolution Capillary Zone Electrophoresis represents an effective tool to assess the diversity of transferrin patterns whereas mass spectrometry is the method of choice to elucidate structural identification about the glycoforms. Hypoglycosylated transferrin glycoforms present in sera of alcohol abusers and normal subjects are structurally identical to those in sera of patients with a congenital disorder of glycosylation type I. Asialo-transferrin, monosialo-transferrin and disialo-transferrin observed in sera of patients with a type II congenital disorder of glycosylation or a hemolytic uremic syndrome, in cerebrospinal fluid and after treatment of serum with neuraminidase are undersialylated transferrin glycoforms with two N-glycans of varying structure. Undersialylated disialo-transferrin is also observed in sera with high levels of trisialo-transferrin
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High-resolution Capillary Zone Electrophoresis for transferrin glycoform analysis associated with congenital disorders of glycosylation.
Wiley-VCH, 2018Co-Authors: Tobler Micha, Caslavsky Jitka, Burda Patricie, Thormann WolfgangAbstract:High-resolution Capillary Zone Electrophoresis is used to assess the transferrin profile in serum of patients with eight different congenital disorders of glycosylation that represent type I, type II and mixed type I/II disorders. Capillary Zone Electrophoresis data are compared to patterns obtained by gel isoelectric focusing. The high-resolution Capillary Zone Electrophoresis method is shown to represent an effective tool to assess the diversity of transferrin patterns. Hypoglycosylated disialo-, monosialo- and asialo-transferrin in type I cases can be distinguished from the corresponding underdesialylated transferrin glycoforms present in type II disorders. The latter can be separated from and are detected ahead of their corresponding hypoglycosylated forms of type I patients. Both types of glycoforms are detected in sera of mixed type I/II patients. The assay has the potential to be used as screening method for congenital disorders of glycosylation. It can be run with a few μL of serum when microvials are used. This article is protected by copyright. All rights reserved
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High-resolution Capillary Zone Electrophoresis for transferrin glycoform analysis associated with congenital disorders of glycosylation
Wiley-Blackwell Publishing Inc., 2018Co-Authors: Tobler Micha, Caslavsky Jitka, Burda Patricie, Thormann WolfgangAbstract:High-resolution Capillary Zone Electrophoresis is used to assess the transferrin profile in serum of patients with eight different congenital disorders of glycosylation that represent type I, type II, and mixed type I/II disorders. Capillary Zone Electrophoresis data are compared to patterns obtained by gel isoelectric focusing. The high-resolution Capillary Zone Electrophoresis method is shown to represent an effective tool to assess the diversity of transferrin patterns. Hypoglycosylated disialo-, monosialo-, and asialo-transferrin in type I cases can be distinguished from the corresponding underdesialylated transferrin glycoforms present in type II disorders. The latter can be separated from and detected ahead of their corresponding hypoglycosylated forms of type I patients. Both types of glycoforms are detected in sera of mixed type I/II patients. The assay has the potential to be used as screening method for congenital disorders of glycosylation. It can be run with a few microliters of serum when microvials are used
