Abstract
Diagnosis of oncological diseases remains at the forefront of current medical research. Carbonic Anhydrase IX (CA IX) is a cell surface hypoxia-inducible enzyme functionally involved in adaptation to acidosis that is expressed in aggressive tumors; hence, it can be used as a tumor biomarker. Herein, we propose a nanoscale graphene oxide (GO) platform functionalized with magnetic nanoparticles and a monoclonal antibody specific to the CA IX marker. The GO platforms were prepared by a modified Hummers and Offeman method from exfoliated graphite after several centrifugation and ultrasonication cycles. The magnetic nanoparticles were prepared by a chemical precipitation method and subsequently modified. Basic characterization of GO, such as the degree of oxidation, nanoparticle size and exfoliation, were determined by physical and chemical analysis, including X-ray photoelectron spectroscopy (XPS), transmission electron microscopy (TEM), energy dispersive X-ray analysis (EDX), and atomic force microscopy (AFM). In addition, the size and properties of the poly-L-lysine-modified magnetic nanoparticles were characterized. The antibody specific to CA IX was linked via an amidic bond to the poly-L-lysine modified magnetic nanoparticles, which were conjugated to GO platform again via an amidic bond. The prepared GO-based platform with magnetic nanoparticles combined with a biosensing antibody element was used for a hypoxic cancer cell targeting study based on immunofluorescence.
Highlights
Carbon-based nanoparticles have attracted a great amount of attention due to their interesting surface properties and ability to act as a platform for modification with various substances, including inorganic nanoparticles, drugs and ligands
The purity and degree of graphene oxide (GO) oxidation were characterized by X-ray photoelectron spectroscopy (XPS) method, while the morphologies and particle size of the GO platforms were examined by atomic force microscopy (AFM) and transmission electron microscopy (TEM)
Graphene oxide platforms were prepared with a mean size of 302 ± 183 nm, as determined by AFM
Summary
Carbon-based nanoparticles have attracted a great amount of attention due to their interesting surface properties and ability to act as a platform for modification with various substances, including inorganic nanoparticles, drugs and ligands. With its numerous exceptional properties, has a wide range of applications in different fields, as has been described by a number of research groups [4,5,6,7,8,9]. The hydrophilicity of GO is a useful property in bioapplications, and numerous different functionalizations with various molecules have been reported [15,16] for the use of modified GO in biosensors, drug carriers or tissue engineering. Factors affecting the cytotoxic effects of GO have been described, such as the lateral dimension, the thickness of the nanolayers, the functional group, and especially the concentration [7,18,19,20]
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