Global ETD Search

1	Production of monoclonal antibodies to sugarcane yellow leaf virus using recombinant read-through protein. Coates, David, Danks, C., Korimbocus, J., Preston, S., Boonham, N., Barker, I. 21 July 2009 (has links) No / Yellow leaf syndrome (YLS) of sugarcane is associated with sugarcane yellow leaf virus (SCYLV), a member of the family Luteoviridae. A fragment of the coat protein and readthrough domain of SCYLV was expressed in a bacterial expression system. The resulting protein was purified and used to immunize mice for monoclonal antibody (MAb) production. Two MAbs, 3A2E3 and 2F7H5, were selected following the screening of hybridoma cells using both plate-trapped antigen enzyme-linked immunosorbent assay (PTA-ELISA) and tissue blot immunoassay (TBIA). These MAbs can be incorporated into the TBIA assay currently used for the routine detection of SCYLV but could not be used in triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA). The two antibodies selected have slightly different specificities. Antibody 3A2E3 gave equivalent results to a polyclonal antiserum (raised to purified virus) in comparative testing using TBIA. The MAbs produced should provide a widely available, uniform reagent for SCYLV diagnosis with the potential to help manage YLS. Vertebrata Mammalia Rodentia Virus Diagnostic Technique ELISA Hybridome Virus recombinant Anticorps monoclonal Sugarcane yellowleaf Luteoviridae
2	Produktion von monoklonalen Antikörpern und Phagenantikörpern gegen das Rinder-Prionprotein durch SFV Partikel-vermittelte Immunisierung von PrP0/0-Mäusen / Production of monoclonal and phage antibodies against bovine prion protein in PrP0/0 mice with the help of recombinant SFV particles Ahmad-Omar, Omar 26 October 2001 (has links) No description available. 570 Biowissenschaften, Biologie Mathematics and Computer Science Prionprotein monoklonale Antikörper Phagenantikörper ScFv nCJD BSE SFV PrP0/0-Mäuse Hybridome prion protein monoclonal Antibodies phagen antibodies ScFv nCJD BSE SFV PrP0/0 mice Hybridoma 42.13 Molekularbiologie WD
3	Production d’un anticorps monoclonal anti-Dal pour le typage sanguin canin Corrales Mesa, Cindy Lizbet 04 1900 (has links) Étant donné l'immunogénicité de l’antigène Dal et sa prévalence élevée (> 98% des chiens sont Dal-positifs), il peut être extrêmement difficile de trouver du sang compatible pour un patient Dal-négatif précédemment transfusé et ayant besoin d’une deuxième transfusion sanguine. De plus, l’accès aux réactifs pour le typage sanguin est actuellement limité, notamment parce qu'il dépend d’anticorps polyclonaux (AcP) produits à la suite de la sensibilisation de trois rares chiens Dal-négatifs identifiés sporadiquement au cours de la dernière décennie dans la colonie d'enseignement de la Faculté de médecine vétérinaire de l'Université de Montréal. Par conséquent, l’objective de cette étude était de produire et de caractériser un anticorps monoclonal murin (AcM) dirigé contre l’antigène canin Dal afin d’assurer la pérennité du typage sanguin Dal. Utilisant la technologie conventionnelle des hybridomes, 5 souris femelles BALB/c ont été immunisées par des injections intrapéritonéales répétées avec des concentrés de globules rouges canines lavés (GRc) Dal-positifs jusqu'à ce que le titrage d’anticorps soit suffisant (> 1 : 10000). Après la fusion de cellules spléniques avec des cellules de myélome, 573 surnageants ont été récoltés à partir du jour 12 post-fusion pour le dépistage avec la technique d'agglutination sur colonne de gel utilisant des GRc Dal-négatif et Dal-positif connus. Parmi 15 surnageants qui ont montré une réaction d’agglutination, un seul avait le patron souhaité (c'est-à-dire anti-Dal). Afin d’évaluer la spécificité et la sensibilité de l’AcM, le typage sanguin Dal de 62 échantillons de GRc a été réalisé en utilisant le AcM anti-Dal et deux AcP canins préalablement caractérisés: 45 échantillons Dal-positifs et 17 Dal-négatifs ont été identifiés avec une concordance de 100 % entre les réactifs (kappa = 1). L’AcM anti-Dal produit a été déterminé comme étant une IgG1. / Given the immunogenicity of the Dal antigen and its high prevalence (>98% of dogs are Dal-positive), it can be extremely difficult to find compatible blood for a previously transfused Dal-negative patient in need of a second blood transfusion. Moreover, access to blood typing reagents is currently limited, in part because it relies on polyclonal antibodies (PAb) produced following the sensitization of three rare Dal-negative dogs identified sporadically over the last decade in the teaching colony of the Faculty of Veterinary Medicine of the University of Montreal. Therefore, the objective of this study was to produce and characterize a murine monoclonal antibody (MAb) directed against the canine Dal antigen, to ensure perennity for Dal blood typing. Using conventional hybridoma technology, 5 BALB/c female mice were immunized by repeated intraperitoneal injections with Dal-positive washed canine red blood cell (cRBC) concentrates until antibody titer was sufficient (>1 : 10000). After fusion of splenic cells with myeloma cells, 573 supernatants were collected starting 12 days post-fusion for screening with the gel column agglutination technique using known Dal-negative and Dal-positive cRBC. Among 15 supernatants that showed an agglutination reaction, only one had the desired pattern (i.e., anti-Dal). To assess the specificity and sensitivity of the MAb, the Dal blood typing of 62 cRBC samples was performed using the anti-Dal MAb and two previously characterized canine PAb: 45 Dal-positive and 17 Dal-negative samples were identified with 100% agreement between reagents (kappa = 1). The anti-Dal MAb produced was determined to be IgG1. Anticorps monoclonal antigène érythrocytaire canin technique sur colonne de gel hybridome groupe sanguin Monoclonal antibody Dog erythrocyte antigen Gel column technique Hybridoma Blood type
4	Immunochemical and chromatographic methods for two anthropogenic markers of contamination in surface waters Carvalho, Jose Joao 08 December 2011 (has links) Koffein (1,3,7-Trimethylxanthin) und Coprostanol (5beta-cholestan-3beta-ol) wurden im Berliner Oberflächenwasser nachgewiesen. Ihre Konzentrationen korrelierten mit dem Verunreinigungsgrad der Proben, was nahelegt, dass sie sich als Marker für menschliche Aktivität eignen. Bemerkenswerterweise wurde Koffein in jeder einzelnen Oberflächenwasserprobe oberhalb der Bestimmungsgrenze von 0,025 µg/L gefunden. Um Oberflächenwasserproben in größeren Serien zu untersuchen, war die Entwicklung zweier neuer Methoden erforderlich: ein Immunoassay, basierend auf einem monoklonalen Antikörper für Koffein und eine dispersive flüssig-flüssig Mikroextraktionsmethode (DLLME), gefolgt von Flüssigkeitschromatographie gekoppelt mit Tandem-Massenspektrometrie (LC-MS/MS) für Coprostanol. Der entwickelte Koffein-Immunoassay zeigt die beste je erhaltene Nachweisgrenze für Koffein (0,001 µg/L), erlaubt Hochdurchsatz-Analysen und erfordert keine Probenvorbereitung. Der Assay wurde auch erfolgreich für die Messung von Koffein in Getränken, Haarwaschmitteln, Koffeintabletten und menschlichem Speichel angewendet. Antikörper gegen Coprostanol sind nicht kommerziell erhältlich. Eine neue Strategie Anti-Coprostanol-Antikörper zu generieren wurde erarbeitet, die eine analoge Verbindung – Isolithocholsäure (ILA) – als Hapten verwendet, mit der eine Gruppe von Mäusen immunisiert wurde. Ein polyklonales Anti-ILA-Serum wurde produziert, welches Coprostanol bindet, aber die niedrige Affinität erlaubte nicht den Aufbau eines Immunoassays, der die Messung von Umweltkonzentrationen des Anayten (im Bereich ng/L) zulässt. Spezifische Anti-ILA-Immunglobuline G wurden auch in den Faeces der Mäuse gefunden. Coprostanol wurde in den Wasserproben durch die Verwendung einer neuentwickelten LC-MS/MS-Methode unter APCI-Ionisation (atmospheric pressure chemical ionisation) gemessen. Konzentrationen oberhalb von 0,1 µg/L wurden nach Voranreicherung der Probe mittels DLLME bestimmt. / Caffeine (1,3,7-trimethylxanthine) and coprostanol (5beta-cholestan-3beta-ol) were detected in samples of Berlin’s surface water. Their concentrations correlated with the contamination status of the samples, suggesting their usefulness as markers of human activity. Remarkably, caffeine concentrations were always well above the limit of quantitation of 0.025 µg/L. In order to screen surface water samples in larger series, the development of two novel methods was required: a monoclonal antibody-based immunoassay for caffeine and a dispersive liquid-liquid microextraction (DLLME) method, followed by liquid chromatography tandem mass spectrometry (LC-MS/MS) for coprostanol. The caffeine immunoassay developed shows the best analytical limit of detection (LOD) obtained so far for caffeine (0.001 µg/L), allows high-throughput analysis, and does not require sample pre-treatment. The assay was also successfully employed to measure caffeine in beverages, shampoos, caffeine tab-lets, and human saliva. Antibodies to coprostanol are not commercially available. A new strategy to generate anti-coprostanol antibodies was elaborated using an analogous com-pound as hapten – isolithocholic acid (ILA) – and immunizing a group of mice. A polyclonal anti-ILA serum was produced, which binds coprostanol but the low affinity did not permit setting up an immunoassay to measure environmental concentrations of the analyte (in the range of ng/L). Specific anti-ILA immunoglobulin G were also found in the faeces of the immunized mice. Coprostanol was quantified in the water samples using a newly developed LC-MS/MS method using atmospheric pressure chemical ionisation (APCI). Concentrations above 0.1 µg/L were determined after sample preconcentration using DLLME. This extraction method also proved to be successful for enrichment of coprostanol-related compounds such as cholesterol, cholestanol, cholestanone, ergosterol, and stigmasterol. HPLC Antikörper Cholesterol ELISA LC-MS/MS Koffein Coprostanol Oberflächenwasser Marker für menschliche Aktivität Immunoassay monoklonalen Antikörper Speichel Isolithocholsäure ILA polyklonales Antikörper Umweltchemie Immunglobuline G hybridome Cholestanol Cholestanon Ergosterol Stigmasterol HPLC ELISA antibodies monoclonal antibody LC-MS/MS water Caffeine coprostanol surface water markers of human contamination Immunoassay saliva isolithocholic acid ILA bile acids polyclonal antibodies environmental chemistry pollution immunoglobulin G hybridoma Cholesterol Cholestanol Cholestanone Ergosterol Stigmasterol 540 Chemie 30 Chemie VK 8547 VK 8627 VN 9367 ddc:540

1

Page generated in 0.027 seconds