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Anal Biochem ; 543: 55-61, 2018 02 15.
Artículo en Inglés | MEDLINE | ID: mdl-29221707

RESUMEN

Rapid diagnosis of Lyme borreliosis has been carried out on chemically modified porous polyethylene sinter bodies. Photografting of 2-propenol on sinter body's surface was performed as a first step, introducing active hydroxyl groups as a result of polyalcohol formation. The hydroxyl groups were used for further immobilization and could be linked via 3-aminopropyltriethoxysilane (APTES) to polysaccharides like mannan. Prone to coupling, mannan was activated using N, N'-disuccinimidyl carbonate (DSC) to allow smooth reaction with the primary amine groups of the silane layer. In a final preparation step, a recombinant fusion protein consisting of the mannan-binding domain of the lectin Concanavalin A (ConA) and a specific Borrelia surface antigen was immobilized by self-organization on the mannan surface. The fusion protein was used as biological interface structure. This strategy is highly efficient and resulted in a defined orientation of the antigen part of the fusion protein. Rapid and convenient differentiation could be then established between Borrelia-negative and a -positive serum even in 1000-fold diluted samples and detection of Lyme borreliosis in a rather early stage is likely. Furthermore, this generic strategy can be easily transferred to other bacterial or viral antigen structures.


Asunto(s)
Antígenos Bacterianos/genética , Borrelia/aislamiento & purificación , Enfermedad de Lyme/diagnóstico , Mananos/química , Polietileno/química , Antígenos Bacterianos/química , Antígenos Bacterianos/inmunología , Borrelia/inmunología , Concanavalina A/química , Tamaño de la Partícula , Porosidad , Proteínas Recombinantes de Fusión/química , Propiedades de Superficie
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