{"id":98638,"date":"2018-03-11T10:19:30","date_gmt":"2018-03-11T10:19:30","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/nuevos-metodos-para-la-inmovilizacion-orientada-de-anticuerpos-sobre-soportes-solidos\/"},"modified":"2018-03-11T10:19:30","modified_gmt":"2018-03-11T10:19:30","slug":"nuevos-metodos-para-la-inmovilizacion-orientada-de-anticuerpos-sobre-soportes-solidos","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/biologia-molecular\/nuevos-metodos-para-la-inmovilizacion-orientada-de-anticuerpos-sobre-soportes-solidos\/","title":{"rendered":"Nuevos metodos para la inmovilizacion orientada de anticuerpos sobre soportes solidos"},"content":{"rendered":"<h2>Tesis doctoral de <strong> Pilar Batalla Bosquet <\/strong><\/h2>\n<p>Immobilized antibodies have a broad variety of uses and a great potential in areas as  immunoassays, biosensors and immunoaffinity chromatography, due to their high sensitivity and high  specificity.  However, it is  very important to know how the antibodies are attached to the  support\u00c2\u00bfs  surface in order to determine if the antigen binding sites of the antibody are accessible to the antigen.  the influence of an antibody\u00c2\u00bfs orientation on its activity has been broadly studied, so it has been proved  that a random antibody orientation diminishes its activity. Usually it is desired to have the antibody  bound to the surface through the fc region instead fab, where the antigen binding site is. Nevertheless,  there is not an ideal method for coupling the antibody. We can find a wide variety of methods for  antibody coupling, and we should choose the method that fits better to our desired application. for these reasons we have developed several methodologies that allow us to bind the antibody  through different areas and to different kinds of surfaces: agarose-amine-aspartic, boronate-glyoxyl,  ida-cu 2+ -glyoxyl, anea-epoxy, cm-epoxy, cm-glyoxyl,  glyoxyl-agarose. With all these supports we  have studied the kinetics of the immobilization, the amount of bound antibody, the orientation of the  antibody, and the activity that the antibody present after immobilization  \u00c2\u00bf that is, how much antigen a  molecule of antibody can bind. So, depending of the conjugate,  we could have between 1.78 to 12.5  nanomols bound antibody\/g support while the activity, expressed as mol antigen\/mol antibody, between  0.6 to 1.6. moreover, we wanted to be sure that the supports that we were going to work with were unable  to react non-specifically with the proteins that we wanted to detect. Non-specific adsorption can affect  the sensitivity of an immunoaffinity column and they can give false positives when using the immobilized  antibodies  as a biosensor. So, once the antibody is immobilized, we have designed also different  strategies to block the conjugates, preventing the non-desired protein adsorptions. it has been also proposed a new approach to bind antibodies to high hydrophobic surfaces, via  protein a. This protein could bound to modified lipases that were adsorbed onto hydrophobic surfaces,  and afterwards the antibody would react with the protein a. The adsorption mechanism of the lipase  permitted us to desorpt the bioconjugate (lipase+protein a+antibody) whenever we wanted without  disrupting the conjugate. Later, this chimeric protein could bound to another hydrophobic support  \u00c2\u00bffor  example, magnetic nanoparticles- through the same mechanism. finally, it has been described the different applications of the immobilization strategies  depending of the orientation of the antibody. For example, some supports will be more adequate for  immunoassays, while others can be better applied in biosensors.<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Nuevos metodos para la inmovilizacion orientada de anticuerpos sobre soportes solidos<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Nuevos metodos para la inmovilizacion orientada de anticuerpos sobre soportes solidos <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Pilar Batalla Bosquet <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Aut\u00f3noma de Madrid<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 15\/01\/2010<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h3>Direcci\u00f3n y tribunal<\/h3>\n<ul>\n<li><strong>Director de la tesis<\/strong>\n<ul>\n<li>Roberto Fern\u00e1ndez Lafuente<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: Mar\u00eda  cruz Moreno bondi <\/li>\n<li>Jes\u00fas Mart\u00ednez de la fuente (vocal)<\/li>\n<li>cesar Mateo gonzalez (vocal)<\/li>\n<li>victor Manuel Fernandez lopez (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Pilar Batalla Bosquet Immobilized antibodies have a broad variety of uses and a great potential in areas [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"default","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center 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