Tratamientos de colummnas capilares de sílice fundida para la obtención de capilares funcionalizados con grupos vinilo

Referencia OEPM: P9800306.-- Fecha de solicitud: 16/02/1998.-- Titular: Consejo Superior de Investigaciones Científicas (CSIC).Tratamientos de columnas capilares de sílice fundida para la obtención de capilares funcionalizados con grupos vinilo. Los capilares empleados para EC y CLCA consisten normalmente en tubos de sílice fundida debido a sus buenas propiedades en términos de resistencia mecánica y transparencia a la radiación ultravioleta-visible. En el caso de EC, la buena conductividad térmica de la sílice fundida ha sido además una propiedad determinante para su empleo generalizado. Estos capilares una vez funcionalizados son recubiertos de un polímero de modo que gracias a esta interacción covalente pared-polímero se asegura la durabilidad y resistencia tanto mecánica como química del recubrimiento.Peer reviewe

Incorporación de átomos de nitrógeno en la superficie del vidrio pyrex y su aplicación a la cromatografía de gases

Tesis inédita de la Universidad Complutense de Madrid, Facultad de Ciencias Químicas, leída en 1977.Fac. de Ciencias QuímicasTRUEProQuestpu

Procedimiento para el análisis de isoformas de glicoproteinas por electroforesis capilar

La presente invención se refiere a un nuevo procedimiento para la purificación, concentración, separación y determinación de las isoformas de la α-1-glicoproteína ácida (AGP), en muestras de suero sanguíneo humano, por electroforesis capilar. El nuevo procedimiento se basa en la inmunocaptura y preconcentración de la muestra dentro del capilar de separación, mediante el uso de una fase inmunoadsorbente magnéticamente inmovilizada en el interior del capilar de electroforesis, y la posterior desorción y separación de las isoformas de la glicoproteína tras la inducción de un efecto de enfoque. El nuevo procedimiento puede tener aplicaciones para el uso de la AGP como biomarcador de enfermedades tales como cáncer, enfermedades vasculares y enfermedades inflamatorias.Peer reviewedConsejo Superior de Investigaciones Científicas (España)A1 Solicitud de patente con informe sobre el estado de la técnic

Fabricación de columnas capilares internamente recubiertas de polímeros para electroforesis capilar

Referencia OEPM: P9701655.-- Fecha de solicitud: 28/07/1997.-- Titulares: Sugelabor, S.A., Consejo Superior de Investigaciones Científicas (CSIC).Fabricación de columnas capilares internamente recubiertas de polímeros para electroforesis capilar. El desarrollo de la Electroforesis Capilar como técnica analítica, ha supuesto la posibilidad de su utilización en un gran número de nuevas aplicaciones. Algunas de estas aplicaciones, sobre todo en el campo de la biotecnología, suponen el uso de columnas capilares que garanticen el control del flujo electroosmótico, especialmente por dar lugar a flujo electroosmótico prácticamente nulo y flujo electroosmótico invertido, eliminando al mismo tiempo los efectos de adsorción entre las sustancias a analizar y la pared interna del capilar. En esta patente se propone un método para la obtención de columnas capilares recubiertas con polímeros que aseguran estos dos supuestos, tratándose de polímeros neutros hidrófilos en el primero y polímeros con carga positiva en el segundo. El método asegura la reproducibilidad de los resultados analíticos derivados de su uso.Peer reviewe

Development of a method for quantitative analysis of the major whey proteins by capillary electrophoresis with on-capillary derivatization and laser-induced fluorescence detection

The main whey proteins have been derivatized on-capillary with 3-(2-furoyl)quinoline- 2-carboxaldehyde (FQ) and analyzed using a laboratory-made capillary electrophore- sis apparatus provided with a laser-induced fluorescence detector. Several param- eters controlling on-capillary derivatization of proteins, including pH, mixing time, reaction time, concentration of the reagents (potassium cyanide and FQ), and reac- tion temperature, were optimized. Coefficient variations were lower than 1% for migration time and 7% for peak height. Assay detection limits for the different proteins were in the range 5 nM to 10 nM. The method developed was applied to the separa- tion of the major whey proteins in a laboratory-made cheese whey and in an infant food formulated with milk. In addition, the b-LG content of these samples was quanti- tated. The results showed good agreement with those given by an RP-HPLC method and with those reported in the literature.M.T.V. acknowledges the Spanish Ministry of Education and Science for a pre-doctoral fellowship. This work has been supported by CICYT (TIC2003-01906 project). Our thanks go to Hero Espa a S.A., especially to Dr. I. Vasal- lo, for the infant formulas and the information provided. We also acknowledge Dr. Olano and his research group (IFI, Spain), especially E. Casal, for the cheese-whey and its RP-HPLC analysis.Peer reviewe

On-capillary derivatization and analysis of amino acids in human plasma by capillary electrophoresis with laser-induced fluorescence detection: Application to diagnosis of aminoacidopathies

A capillary electrophoresis with laser-induced fluorescence detection method for the analysis of free amino acids (AA) in human plasma was developed. A mixture of 16 AA was on-capillary derivatized with 3-(2-furoyl)quinoline-2-carboxaldehyde (FQ) and separated inside the capillary in less than 30 min using 70 mM borax-3.5 mM SDS pH 9.3 as running buffer. Four plasma samples from a healthy donor and patients suffering from phenylketonuria, propionic acidemia, and tyrosinemia type II were studied. Repeatabilities calculated as intra-day RSD (n = 3) values for the AA involved in these aminoacidopathies (glycine, phenylalanine, and tyrosine) were in the range of 0.3 to 1.2% for migration time and 3.7 to 8.2% for peak height. Reproducibilities calculated as inter-day RSD (n = 4) values for the same AA were between 0.7 and 1.4% for migration time and 4.7 and 9.1% for peak height. A fast qualitative analysis allowed the identification of the corresponding disease by comparing the electrophoretic profiles from the patient and the healthy donor and noting the increased level of the specific AA accompanying each individual disease. The results of the quantitative analysis for glycine, phenylalanine, and tyrosine in the plasma samples studied using the developed method showed a good agreement with those provided by the Center of Diagnosis of Molecular Diseases using a standard method for AA analysis. © 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.Peer Reviewe

On-chip single column transient isotachophoresis with free zone electrophoresis for preconcentration and separation of α-lactalbumin and β-lactoglobulin

Supplementary data to this article can be found online at http://dx. doi.org/10.1016/j.microc.2017.04.040Isotachophoresis (ITP) coupled to zone electrophoresis (ZE), either free zone electrophoresis (FZE) or gel electrophoresis (GE), carried out mainly in capillaries and, although less frequently, also in microchips, is a powerful preconcentration and separation technique which has been successfully used for the study of many low molecular-weight analytes. However, this analytical technique has been scarcely applied for proteins separation. In this work, an on-chip transient ITP coupled to free zone electrophoresis (t-ITP-MFZE) mode with LIF detection is developed for the preconcentration and separation of the proteins α-lactalbumin and β-lactoglobulin. Firstly, for LIF detection, the proteins were off-chip fluorescently labeled with the fluorogenic reagent Chromeo P503. Then, several separation parameters in t-ITP-MFZE mode such as leading electrolyte, terminating electrolyte, separation voltage, and injection time were optimized to achieve the maximum sensitivity while maintaining an adequate resolution between α-lactalbumin and β-lactoglobulin in a single column configuration t-ITP. Using the optimized electrolytes (50 mM imidazole/HCl pH = 8 as leading electrolyte and 100 mM imidazole/12 mM HEPES pH = 8 as terminating electrolyte) separation of both proteins was achieved in less than 4 min with peak resolution of 1.5. The LODs were 55 nM and 380 nM, for α-lactalbumin and β-lactoglobulin, respectively, which are adequate for some food allergenicity studies. Finally, comparison of the optimized t-ITP-MFZE method to the equivalent MFZE method, carried out also in microchips but without the isotachophoretic preconcentration step, provided preconcentration indexes for both proteins around 10.The authors acknowledge the SpanishMinistry of Economy, Industry and Competitiveness (MINECO CTQ2013-43236-R) and CSIC (joint project 2009JP003 with the Japanese Society for Promotion of Science) for financial support. A.G.C. acknowledges CSIC for a JAE-Doc contract. This contract was co-financed by the European Union under the European Social Fund (ESF).Peer Reviewe

Procedimiento para el análisis de isoformas de glicoproteínas por electrofóresis capilar

[EN] The present invention relates to a new method for the purification, concentration, separation and determination of the isoforms of alpha-1-acid glycoprotein (AGP) in human blood serum samples using capillary electrophoresis. The new method is based on the immunocapture and preconcentration of the sample within the separation capillary by using an immunoadsorbent phase magnetically immobilized within the electrophoresis capillary and the subsequent desorption and separation of the glycoprotein isoforms after induction of a focussing effect. The new method may apply to the use of AGP as a biomarker in diseases such as cancer, vascular diseases and inflammatory diseases.[ES] La presente invención se refiere a un nuevo procedimiento para la purificación, concentración, separación y determinación de las isoformas de la α-1-glicoproteína ácida (AGP), en muestras de suero sanguíneo humano, por electroforesis capilar. El nuevo procedimiento se basa en la inmunocaptura y preconcentración de la muestra dentro del capilar de separación, mediante el uso de una fase inmunoadsorbente magnéticamente inmovilizada en el interior del capilar de electroforesis, y la posterior desorción y separación de las isoformas de la glicoproteína tras la inducción de un efecto de enfoque. El nuevo procedimiento puede tener aplicaciones para el uso de la AGP como biomarcador de enfermedades tales como cáncer, enfermedades vasculares y enfermedades inflamatorias.Peer reviewedConsejo Superior de Investigaciones CientíficasA1 Solicitud de patente con informe sobre el estado de la técnic

On-line immunoaffinity capillary electrophoresis based on magnetic beads for the determination of alpha-1 acid glycoprotein isoforms profile to facilitate its use as biomarker

An immunoaffinity purification method coupled on-line to capillary electrophoresis (IACE) which allows the determination of several isoforms of intact alpha-1 acid glycoprotein (AGP) in serum samples using UV detection is developed. The immunoaffinity step is based on anti-AGP antibodies (Abs) covalently bound to magnetic beads (MBs) which are captured at the inlet end of the capillary using permanent magnets placed inside the cartridge of the CE instrument. The on-line method includes injection of the MBs with the Ab bound (MBs-Ab) and their trapping by the magnets at the entrance of the separation column, injection of serum sample and capture of AGP by the Abs, release of captured AGP, focus of desorbed protein, separation of AGP isoforms, and removal of MBs-Ab. The optimization of the different factors involved in each step allowed purification, separation and detection of AGP isoforms in a single electrophoretic analysis in about 1. h. Automation, sample and reagents consumption as well as analysis time was improved compared to off-line alternatives which use purification of AGP in an immunochromatographic column and CE separation of AGP isoforms in two independent operations. The analytical methodology developed allows the separation of 10 AGP isoforms in serum samples from a healthy donor. For a serum sample, precision (expressed as relative standard deviation) in terms of corrected area percentage was better than 0.5% for each peak accounting for more than 10% of total AGP and it was better than 4.0% in terms of relative migration time of each AGP isoform considering the whole process. © 2013 Elsevier B.V.Peer Reviewe