Development of a borreliosis assay: Mannan coated polyethylene sinter bodies as a new platform technology
| Autor: | Nina Dassinger, Doru Vornicescu, Michael Keusgen, Mohammed Alasel |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Surface Properties Recombinant Fusion Proteins 030106 microbiology Biophysics Polysaccharide Biochemistry Mannans 03 medical and health sciences Blood serum Antigen Concanavalin A Particle Size Molecular Biology Mannan chemistry.chemical_classification Antigens Bacterial Lyme Disease biology Borrelia Lectin Cell Biology bacterial infections and mycoses Combinatorial chemistry Fusion protein chemistry Polyethylene Photografting biology.protein Porosity |
| Zdroj: | Analytical biochemistry. 543 |
| ISSN: | 1096-0309 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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