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Oriented functionalization of magnetic nanoparticles with cadherins

AuthorsMoreno Antolín, Eduardo ; Martín-Gracia, Beatriz; Fuente, Jesús M. de la; Fratila, Raluca M.; Grazú, Valeria
Issue Date2018
Citation12th International Conference on the Scientific and Clinical Applications of Magnetic Carriers (2018)
AbstractMagnetic nanoparticles (MNPs) have shown a great potential for biomedical applications due to their stability, biocompatibility and superparamagnetic behaviour below 20 nm. Thus, they are widely used in biosensing, magnetic hyperthermia, or controlled drug delivery, to name but a few applications. Specifically, in order to achieve active targeting systems for therapy, MNPs are frequently vectorized by means of their biofunctionalization with biomolecules. However, there is no universal MNPs biofunctionalization protocol; large differences among MNP types and their coatings and the vast diversity of biomolecules to be attached are factors that difficult this critical step. Each particular case requires optimization to find the most adequate method to preserve the biological activity of the targeting biomolecule; for instance, its proper orientation once attached to the MNPs is crucial for maintaining its activity and must be achieved through innovative functionalization methodologies. In this sense, we decided to take advantage of the affinity of chelated metals for histidine residues present in His-tagged biomolecules to develop a protocol for oriented functionalization of MNPs with cadherin recombinant fragments. Our final aim is to selectively attach the cadherinfunctionalized MNPs to cell membranes for localized magnetic hyperthermia studies. Cadherins are a family of Ca2+-dependent cell-cell adhesion proteins with a precise spatio-temporal pattern during embryonic development. Among them, E-cadherin is the most studied, having a relevant role in maintaining the cohesion of epithelial tissue. Moreover, during the epithelialmesenchymal transition, E-cadherin disappearance is associated with metastasis. MNPs were synthesized by thermal-decomposition and covered with an amphiphilic polymer (poly(maleic anhydride-alt-1-octadecene)). Their functionalization with Cu2+ chelated by nitrilotriacetic acid (NTA) via carbodiimide chemistry was optimized. Afterwards, those MNPs were incubated with E/EC1-2-cadherin fragments, allowing the Cu2+-His interaction to take place. The attached protein was quantified from the supernatant. We are currently testing the use of different passivating agents (TRIS, polyethylene glycol) to stabilize the MNPs in cell culture media. The cytotoxicity of the MNPs, as well as preliminary studies of immobilization on the membrane of MDCK cells are also underway.
DescriptionResumen del trabajo presentado a la 12th International Conference on the Scientific and Clinical Applications of Magnetic Carriers, celebrada en Copenhague (Dinamarca) del 22 al 26 de mayo de 2018.
Appears in Collections:(ICMA) Comunicaciones congresos
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