20.500.12556/RUL-129218 Funkcionalizacija monodisperznih delcev silicijevega dioksida s koronavirusnima proteinoma ORF8 in N Functionalization of monodisperse silica particles with coronavirus proteins ORF8 and N Virus SARS-CoV-2 je predstavnik koronavirusov, ki se je pojavil konec leta 2019 in povzročil pandemijo covida-19. Monodiperzni delci silicijevega dioksida v raztopinah kažejo zanimive optične lastnosti, ki so uporabne pri izdelavi biosenzorja, s katerim bi lahko zaznali prisotnost virusa. Eden izmed korakov pri izdelavi biosenzorja je tudi funkcionalizacija delcev z molekulami, ki omogočajo zaznavanje tarčnih biomolekul. V diplomskem delu smo se osredotočili na njegov strukturni protein N in pomožni protein ORF8. Oba interagirata z imunskim sistemom gostitelja in sta močno imunogena. Proteina smo izrazili v bakterijskem sistemu E. coli BL21[DE3]. Iz topne frakcije smo s pomočjo nikljeve afinitetne kromatografije uspeli izolirati le protein N, ki smo mu nato s proteazo TEV odcepili heksahistidinsko oznako in ga dodatno skoncentrirali. Z njim smo funkcionalizirali monodisperzne delce SiO2, ki smo jih sintetizirali na dva različna načina in predhodno aktivirali z aldehidnimi funkcionalnimi skupinami. Pokazali smo, da se je protein vezal na delce, vendar zaradi prevelike občutljivosti testa, s katerim smo preverjali uspešnost funkcionalizacije, nismo uspeli nedvoumno dokazati, da je kovalentna vezava bolj učinkovita od adsorpcije. The SARS-CoV-2 virus is a member of the Coronaviridae family that emerged in late 2019 and caused the COVID-19 pandemic. Monodisperse silica particles have interesting optical properties in solutions that could be used in the design of a biosensor for virus detection. One of the most important steps in biosensor production is the functionalization of the particles with molecules that allow the detection of target molecules. In this thesis, the structural N protein and the accessory protein ORF8 were investigated. Both proteins inhibit the host immune response and are highly immunogenic. The proteins were expressed in E. coli BL21[DE3]. Using nickel affinity chromatography, only protein N was isolated from the soluble fraction. Moreover, the hexahistidine tag was cleaved by TEV protease, and the N protein was additionally concentrated. Monodisperse SiO2 particles were synthesized in two ways, activated with aldehyde functional groups, and functionalized with the N protein. Protein binding to the particles was demonstrated, but the high sensitivity of the assay method used did not provide clear evidence that the covalent coating was better than the adsorption. SARS-CoV-2 silicijev dioksid nanodelci funkcionalizacija SARS-CoV-2 silicium dioxide nanoparticles functionalization true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2021-08-30 13:05:00 2021-08-30 13:05:05 2022-09-08 04:07:19 0000-00-00 00:00:00 2021 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 19461 75977219 Zerjav_Neza_-_Funkcionalizacija_monodisperznih_delcev_silicijevega_dioksida_s_koronavirusnima_pr.pdf Zerjav_Neza_-_Funkcionalizacija_monodisperznih_delcev_silicijevega_dioksida_s_koronavirusnima_pr.pdf 1 C9C05B4A1C145B37946788052FB4BD15 aa71efa9da62c075ef9c6e6a55214edc5ad918252926c515ff39386c4aebb3aa e692234a-0981-11ec-a523-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=146170 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0