000086831 001__ 86831
000086831 005__ 20200108151739.0
000086831 037__ $$aTAZ-TFM-2019-301
000086831 041__ $$aeng
000086831 1001_ $$aCastro Hinojosa, Christian
000086831 24200 $$aMagnetic nanoparticles immobilization on cell membranes mediated by cadherins for magnetic hyperthermia studies
000086831 24500 $$aInmovilización de nanoparticulas magnéticas en membranas celulares mediante cadherinas para estudios de hipertermia magnética
000086831 260__ $$aZaragoza$$bUniversidad de Zaragoza$$c2019
000086831 300__ $$ematerial-fisico-adicional
000086831 506__ $$aby-nc-sa$$bCreative Commons$$c3.0$$uhttp://creativecommons.org/licenses/by-nc-sa/3.0/
000086831 520__ $$aMagnetic nanoparticles (MNPs) have the ability to generate heat when exposed to an alternating magnetic field. In biomedicine this property can be exploited to study the effects of localized heat generation on cell membranes. For this purpose, it is essential to target the MNPs to specific cell membrane receptors and achieve a stable and long-lasting immobilization of the MNPs on the cell membrane. Several molecules can be used as vectors; however, a stable conjugation is often difficult to achieve. Herein we explore the use of E-cadherin membrane protein as a potential targeting tool to generate a stable immobilization of MNPs on the cell membrane, based on homophilic E-cadherin – E-cadherin interactions. MNPs (12 nm diameter) were functionalized with polyethylene glycol (PEG) chains (MW: 750 or 5000 Da) and a derivative of nitrilotriacetic acid (NTA-Cu2+) in a single step using EDC chemistry; then, E-cadherin protein fragments (wild type: WT or mutant: W2A) were conjugated on the surface of the nanoparticles in an oriented manner through their histidine tail (His-tag), generating a protocol that could be easily adapted to any protein containing a His-tag in its structure. The cell-nanoparticle interactions were evaluated in Madin-Darby canine kidney (MDCK) and mouse fibroblast BALB/c3T3 cells, expressing and lacking E-cadherins respectively. The results obtained showed that the MNPs coated with a larger PEG decreased the extent of non-specific interaction with cells. On the other hand, when the MNPs were immobilized by the W2A mutant protein fragments, their interaction with the MDCK cells was lower in comparison with the one of the MNPs functionalized with WT fragments. Furthermore, in the case of BALB/c3T3 cells, lacking E-cadherins on their membranes, the interaction with the different types of MNPs did not show clear differences. These results suggested that the MNPs interact with MDCK cells through homophilic interactions; furthermore, the mutation present in the protein fragments W2A generates changes in the binding of nanomaterials to the cell membrane, which could be used in future studies to improve the specificity and stability of the MNPs immobilization on cell membrane.<br /><br />
000086831 521__ $$aMáster Universitario en Biotecnología Cuantitativa
000086831 540__ $$aDerechos regulados por licencia Creative Commons
000086831 700__ $$aFratila, Raluca Maria$$edir.
000086831 700__ $$aMoros Caballero, María$$edir.
000086831 7102_ $$aUniversidad de Zaragoza$$bQuímica Orgánica$$cQuímica Orgánica
000086831 8560_ $$f790741@celes.unizar.es
000086831 8564_ $$s1478817$$uhttps://zaguan.unizar.es/record/86831/files/TAZ-TFM-2019-301.pdf$$yMemoria (eng)
000086831 8564_ $$s506975$$uhttps://zaguan.unizar.es/record/86831/files/TAZ-TFM-2019-301_ANE.pdf$$yAnexos (eng)
000086831 909CO $$ooai:zaguan.unizar.es:86831$$pdriver$$ptrabajos-fin-master
000086831 950__ $$a
000086831 951__ $$adeposita:2020-01-08
000086831 980__ $$aTAZ$$bTFM$$cCIEN
000086831 999__ $$a20190627210304.CREATION_DATE