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A Validatation of non-invasive method for diagnostic of cystic fibrosis based on ion ratios determined by CE-C4D
Ďurč, P. ; Lačná, J. ; Dosedělová, V. ; Foret, František ; Homoľa, L. ; Pokojová, E. ; Malá, M. ; Skřičková, J. ; Dastych, M. ; Vinohradská, H. ; Dřevínek, P. ; Skalická, V. ; Kubáň, Petr
A validation of novel approach for diagnosis of cystic fibrosis (CF) is presented. The method is very simple and fast, since for collection of samples, only wiping of the skin of forearm is needed. The collected sample is analyzed by capillary electrophoresis with contactless conductometric detection (CE-C4D). Ion ratios are determined as ratios of peak area of chloride anion and peak areas of selected cations. It is possible to distinguish between cystic fibrosis patients and healthy individuals when ion ratio is applied as diagnostic factor. In here, we present analytical parameters of developed method (limits of detection, limits of quantification, repeatability of migration times, repeatability of peak areas) and ROC curve, which reveals high sensitivity and specificity of the developed method.
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Microfluidic device for cell counting and electrical lysis
Václavek, Tomáš ; Křenková, Jana ; Foret, František
This lecture presents two microfluidic devices as essential units for single cell proteomic analysis, firstly a device for cell counting based on resistive pulse sensing as an alternative to optical platforms based on visual control over the sample input, secondly a unit for electrical lysis of cells using direct current voltage pulses to lyse cells one by one with a possibility to analyze the distinct lysate zones by electrospray ionization mass spectrometry.
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Development of Methods for High-Throughput Enrichment of Glycoproteins and Glycopeptides Employing Multiple Lectin Affinity Chromatography/Tandem Mass Spectrometry
Maděra, Milan ; Pacáková, Věra (advisor) ; Feltl, Ladislav (referee) ; Foret, František (referee) ; Kučerová, Zdenka (referee)
5 CONCLUSIONS This thesis surrrnrarizes the development of a multimethodological analýical approach employing microcolumn lectin affi1t1,9fuom1tography coupled to highir".otu,to; separation and dete9tion techniques, in order to facilitate the enrichmení of glyco-proteins aná giycopeptioes originating from small quantities of real samples. This is u"toauy a sipificant-ítep towara satis$'ing the demands of contemporary glycoproteomics, which rely orirurt *a small scale analyses, allowing the identification of low abundant protein components' The overď conclusions and contribution to the contemporaÍy scierrce are drawn in tbree followine chaoters. 5.1 combining Lectin Microcolumns with High-Resolution separation Techniques for Enrichment of Glycoproteins and Glycopeptides This section describes coupling small-scďe lectin aÍfinity chromatograpby on-line to high- resolution separation and detection techniques, with the utiliý of a microcolumn loaded with a lectin immobilized onto macroporous silica. Experimental results, involving optimization of coupling procedure, fabrication of lectin microcolumns, verification of tteir Uin&ng efficiency and their possibility of interfacing on-line with nano LC-MS/]víS, me summarizžd into thé following conclusions; o optimized coupling procedure involved resuspending only 125...
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Enantioselective separation systems in capillary electrophoresis
Svobodová, Jana ; Gaš, Bohuslav (advisor) ; Foret, František (referee) ; Kenndler, Ernst (referee)
Chiral separation systems in capillary electrophoresis are in the scope of interest of many research groups all over the world. Therefore, the need to develop reliable theoretical models, which would help to explain phenomena connected with chiral separations or optimization of separation conditions, is obvious. In this thesis several mathematical models and approaches that can fulfill these requirements are presented. First part of the thesis deals with the determination of rate constants of interconversion of enantiomers by means of dynamic capillary electrophoresis. We focused on mixtures of chiral selectors and formulated a mathematical model, which enables to determine rate constants of interconversion in such systems. Mixtures of chiral selectors are very popular in separation practice due to their enhanced enantioselectivity. The theoretical model established in the thesis is able to explain the separation mechanisms of multi-chiral selector systems and to propose and verify possible ways of their optimization. In addition, the separation mechanism in systems with simultaneous cyclodextrin and borate complexation was revealed. Finally we present the complete mathematical model of electromigration in systems with complexation agents. The model is implemented into our simulation tool Simul 5...
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