• MeSH
• Arginine MeSH
• Electrophoresis MeSH
• Vasopressins isolation & purification   MeSH

• Keywords
• elektroforéza kapilární,

• Conspectus
• Biochemie. Molekulární biologie. Biofyzika
• NML Fields
• fyzika, biofyzika
• biomedicínské inženýrství

Heavy chain disease with monoclonal incomplete mu-heavy chains (mu-HCD) is a rare disorder usually associated with an underlying lymphoproliferative malignancy. Laboratory diagnosis of patients with mu-HCD is usually challenging and the monoclonal protein is not detected by electrophoresis in up to 75% of mu-HCD cases. We describe a patient with multiple malignancies in whom we detected and characterized monoclonal mu-heavy chains using immunofixation electrophoresis, capillary zone electrophoresis with immunotyping, and high resolution two-dimensional electrophoresis. The high resolution 2D electrophoresis enabled us to determine the molecular weight of the mu-heavy chains. The abnormal protein concentration in the serum was unusually high, 38 g/l measured in our patient is the highest reported value in the literature so far.

• MeSH
• Electrophoresis, Gel, Two-Dimensional methods   MeSH
• Electrophoresis, Capillary methods   MeSH
• Middle Aged MeSH
• Humans MeSH
• Lymphoproliferative Disorders diagnosis   MeSH
• Immunoglobulin Heavy Chains analysis   MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Female MeSH

Saccharides form one of the major constituents of biological macromolecules in living organisms. Many biological processes including protein folding, stability, immune response and receptor activation are regulated by glycosylation. In this work, we optimized a capillary electrophoresis method with capacitively coupled contactless conductivity detection for the separation of eight monosaccharides commonly found in glycoproteins, namely D-glucose, D-galactose, D-mannose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, D-fucose, N-acetylneuraminic acid, and D-xylose. A highly alkaline solution of 50 mM sodium hydroxide, 22.5 mM disodium phosphate, and 0.2 mM CTAB (pH 12.4) was used as a background electrolyte in a 10 μm id capillary. To achieve baseline separation of all analytes, a counter-directional pressure of -270 kPa was applied during the separation. The limits of detection of our method were below 7 μg/ml (i.e., 1.5 pg or 1 mg/g protein) and the limits of quantification were below 22 μg/ml (i.e., 5 pg or 3 mg/g protein). As a proof of concept of our methodology, we performed an analysis of monosaccharides released from fetuin glycoprotein by acid hydrolysis. The results show that, when combined with an appropriate pre-concentration technique, the developed method can be used as a monosaccharide profiling tool in glycoproteomics and complement the routinely used LC-MS/MS analysis.

• MeSH
• Acetylgalactosamine MeSH
• Acetylglucosamine MeSH
• Cetrimonium MeSH
• Chromatography, Liquid MeSH
• Electrophoresis, Capillary methods   MeSH
• Electrolytes chemistry   MeSH
• Fetuins MeSH
• Phosphates MeSH
• Fucose MeSH
• Galactose MeSH
• Glucose MeSH
• Glycoproteins chemistry   MeSH
• Sodium Hydroxide MeSH
• N-Acetylneuraminic Acid * MeSH
• Mannose MeSH
• Monosaccharides * analysis   MeSH
• Tandem Mass Spectrometry MeSH
• Xylose MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Application of Tris-N-[Tris(hydroxymethyl)methyl]glycine gels for 2DE is hampered by formation of mixed CHAPS-SDS micelles resulting in typical swirling pattern in the low mass range, which makes reliable quantitative and qualitative gel evaluation impossible. Modification of 2DE strip equilibration procedure prevented the direct interaction between both detergents during equilibration process, thus substantially improving gel separation.

• MeSH
• Electrophoresis, Gel, Two-Dimensional methods   MeSH
• Detergents chemistry   MeSH
• Glycine analogs & derivatives   chemistry   MeSH
• Micelles * MeSH
• Proteome analysis   MeSH
• Proteomics methods   MeSH
• Plant Proteins analysis   MeSH
• Solanum tuberosum chemistry   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

OBJECTIVES: Our aim was to establish a reliable, rapid, and inexpensive method for the simultaneous genotyping of the HMOX-1 (heme oxygenase-1) and UGT1A1 (bilirubin UDP-glucuronosyltransferase) gene promoter variations. RESULTS: The HMOX1 (GT)(n) and UGT1A1 (TA)(n) gene promoter variations were determined by fragment analysis using a single duplex PCR, with different fluorescent dye-labeled primers; followed by multicolored capillary electrophoresis. CONCLUSION: This novel method provides simultaneous genotyping of key tandem repeat variations in the HMOX1 and UGT1A1 promoters.

• MeSH
• Electrophoresis, Capillary methods   MeSH
• Genotype MeSH
• Glucuronosyltransferase genetics   MeSH
• Heme Oxygenase-1 genetics   MeSH
• Polymorphism, Single Nucleotide genetics   MeSH
• Humans MeSH
• Microsatellite Repeats genetics   MeSH
• Reproducibility of Results MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

A simple sample injection procedure compatible with commercial capillary electrophoresis (CE) instrumentation was developed, which enables handling sample volumes as little as 250nL for analytical applications where sample volume availability is of concern. Single-use micro-sampling inserts were prepared by thermal modification of polypropylene micropipette tips and the inserts were accommodated in standard CE vials in CE autosampler carousel. To ensure direct contact of separation capillary injection end with sample solution and to avoid possible damage to the capillary, a soft compression spring was placed at the bottom of the vial underneath the micro-sampling insert. Injections from sub-μL samples were carried out in conventional as well as in short-end injection mode, were compatible with standard i.d./o.d. (25-100μm/365μm) fused silica capillaries and with various background electrolyte solutions and detection modes. Excellent repeatability of replicate injections from 250nL to 3μL was achieved based on RSD values of quantitative analytical measures (peak heights ≤2.4% and peak areas ≤3.7%) for CE-UV-vis, CE-ESI-MS and CE-contactless conductivity detection of model basic drugs. The achieved RSD values were comparable with those for replicate injections of the drugs from standard CE vials. The reported concept of injections from micro-sampling inserts was further demonstrated useful in evaluation of micro-electromembrane extraction (μ-EME) of model basic drugs. Sub-μL volumes of operational solutions resulted in reduced lengths of μ-EME phases and improved extraction recoveries (66-91%) were achieved.

• MeSH
• Electrophoresis, Capillary instrumentation   methods   MeSH
• Electrolytes chemistry   MeSH
• Mass Spectrometry MeSH
• Nortriptyline analysis   isolation & purification   MeSH
• Papaverine analysis   MeSH
• Solutions chemistry   MeSH
• Spectrophotometry, Ultraviolet MeSH
• Publication type
• Journal Article MeSH

Upconversion nanoparticles (UCNPs) are an emerging class of optical materials with high potential in bioimaging due to practically no background signal and high penetration depth. Their excellent optical properties and easy surface functionalization make them perfect for conjugation with targeting ligands. In this work, capillary electrophoretic (CE) method with laser-induced fluorescence detection was used to investigate the behavior of carboxyl-silica-coated UCNPs. Folic acid, targeting folate receptor overexpressed by wide variety of cancer cells, was used for illustrative purposes and assessed by CE under optimized conditions. Peptide-mediated bioconjugation of antibodies to UCNPs was also investigated. Despite the numerous advantages of CE, this is the first time that CE was employed for characterization of UCNPs and their bioconjugates. The separation conditions were optimized including the background electrolyte concentration and pH. The optimized electrolyte was 20 mM borate buffer with pH 8.

• MeSH
• Electrophoresis, Capillary methods   MeSH
• Fluorescent Dyes chemistry   MeSH
• Spectrometry, Fluorescence methods   MeSH
• Folic Acid chemistry   MeSH
• Limit of Detection MeSH
• Linear Models MeSH
• Nanoconjugates chemistry   MeSH
• Antibodies chemistry   MeSH
• Reproducibility of Results MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

In this paper, we demonstrate the effectiveness of a new 3D printed magnet holder that enables capture of magnetic microparticles in commercially available capillary electrophoresis equipment with a liquid or air based coolant system. The design as well as the method to capture magnetic microparticles inside the capillary are discussed. This setup was tested at temperature and pH values suitable for performing enzymatic reactions. To demonstrate its applicability in CE- immobilized microenzyme reactors (IMER) development, human flavin-containing monooxygenase 3 and bovine serum albumin were immobilized on amino functionalized magnetic microparticles using glutaraldehyde. These microparticles were subsequently used to perform in-line capillary electrophoresis with clozapine as a model substrate. This setup could be used further to establish CE-IMERs of other drug metabolic enzymes in a commercially available liquid based capillary coolant system. The CE-IMER setup was successful, although a subsequent decrease in enzyme activity was observed on repeated runs.

• MeSH
• Amines chemistry   MeSH
• Equipment Design instrumentation   MeSH
• Electrophoresis, Capillary instrumentation   MeSH
• Enzymes, Immobilized chemistry   MeSH
• Glutaral chemistry   MeSH
• Clozapine chemistry   MeSH
• Humans MeSH
• Magnetic Fields MeSH
• Magnets chemistry   MeSH
• Microspheres * MeSH
• NADP chemistry   MeSH
• Silicon Dioxide chemistry   MeSH
• Oxygenases chemistry   MeSH
• Surface Properties MeSH
• Serum Albumin, Bovine chemistry   MeSH
• Enzyme Stability MeSH
• Temperature MeSH
• Particle Size MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

An original method based on capillary zone electrophoresis with fluorimetric detection has been developed for the determination of terpenic compounds. The method is based on the separation of a terpenes dynamically labeled by the non-ionogenic tenside poly(ethylene glycol) pyrenebutanoate, which was used previously for the labeling of biopolymers. The background electrolytes were composed of taurine-Tris buffer (pH 8.4). In addition to the non-ionogenic tenside aceton and poly(ethylene glycol) were used as the additives. The capillary zone electrophoresis with fluorometric detection at the excitation wavelength 335 nm and the emission wavelength 463 nm was successfully applied to the analysis of tonalid, cholesterol, vitamin A, ergosterol, estrone and farnesol at level of 10(-17) mol L(-1). Farnesol, is produced by Candida albicans as an extracellular quorum-sensing molecule that influences expression of a number of virulence factors, especially morphogenesis and biofilm formation. It enables this yeast to cause serious nosocomial infections. The sensitivity of this method was demonstrated on the separation of farnesol directly from the cultivation medium.

• MeSH
• Biofilms MeSH
• Butyrates chemistry   MeSH
• Candida albicans chemistry   metabolism   MeSH
• Cholesterol chemistry   MeSH
• Electrophoresis, Capillary methods   MeSH
• Ergosterol chemistry   MeSH
• Estrone chemistry   MeSH
• Farnesol chemistry   isolation & purification   metabolism   MeSH
• Fluorometry methods   MeSH
• Polymers chemistry   MeSH
• Quorum Sensing MeSH
• Sensitivity and Specificity MeSH
• Terpenes chemistry   isolation & purification   MeSH
• Tetrahydronaphthalenes chemistry   MeSH
• Vitamin A chemistry   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH