Catalytically active bovine serum amine oxidase bound to fluorescent and magnetically drivable nanoparticles

Abstract

Novel superparamagnetic surface-active maghemite nanoparticles (SAMNs) characterized by a diameter of 10 ± 2 nm were modified with bovine serum amine oxidase, which used rhodamine B isothiocyanate (RITC) adduct as a fluorescent spacer-arm. A fluorescent and magnetically drivable adduct comprised of bovine serum copper-containing amine oxidase (SAMN-RITC-BSAO) that immobilized on the surface of specifically functionalized magnetic nanoparticles was developed. The multifunctional nanomaterial was characterized using transmission electron microscopy, infrared spectroscopy, mass spectrometry, and activity measurements. The results of this study demonstrated that bare magnetic nanoparticles form stable colloidal suspensions in aqueous solutions. The maximum binding capacity of bovine serum amine oxidase was approximately 6.4 mg g -1 nanoparticles. The immobilization procedure reduced the catalytic activity of the native enzyme to 30% ± 10% and the Michaelis constant was increased by a factor of 2. We suggest that the SAMN-RITC-BSAO complex, characterized by a specific activity of 0.81 IU g -1, could be used in the presence of polyamines to create a fluorescent magnetically drivable H 2 O 2 and aldehydes-producing system. Selective tumor cell destruction is suggested as a potential future application of this system.

In vivo delivery, pharmacokinetics, biodistribution and toxicity of iron oxide nanoparticles

“…175 There are some recent methods reported for developing colloidally stable un-coated IONPs. 176-179 However, further studies are required to evaluate the in vivo performance of these IONPs formulations. Different types of natural ( e.g.…”

Nanotechnology-Based Strategies to Develop New Anticancer Therapies

“…The surface coating of nanoparticles is fundamental to produce physically and chemically stable colloidal systems and to provide functional groups allowing conjugation of active molecules and or targeting ligands. Stabilization of IONPs can be accomplished by several processes [51], such as i) surface derivatization with polymeric stabilizers and/or surfactants (e.g., dextran, polyvinyl-alcohol, polyethylene-glycol) or by deposition of thin layers of either inorganic metals (gold), nonmetals (b) SAMN@RITC-BSAO, in which the enzyme was immobilized on the surface of magnetic iron oxide nanoparticles through a fluorescent spacer arm [47]; (c) Au@pDMPA/HCl-BSAO, a core-shell gold nanoparticles stabilized with the hydrophilic polymer (poly(3-dimethylammonium-1-propyne hydrochloride) and decorated with BSAO as described in the main text [45].…”

“…Amino acids, containing chelating functionalities as imidazole or carboxylic groups, are prone to coordinate transition metals such as Ni(II), Cu(II), Zn(II), and Co(II). This property is commonly applied for protein purification [138] and can be used for biomolecule immobilization on metal and metal oxide nanoparticles [47]. Genetically encoded poly-histidine tags (His-tags), usually consisting of six sequential histidine residues, is able to chelate transition metals, such as Cu(II), Co(II), Zn(II), or Ni(II).…”

“…Exploiting different nano-based delivery strategies, bovine serum amine oxidase (BSAO) has been used to kill tumor cells. In the cartoon, two different BSAO smart nano-vehicles are depicted:(b) SAMN@RITC-BSAO, in which the enzyme was immobilized on the surface of magnetic iron oxide nanoparticles through a fluorescent spacer arm[47]; (c) Au@pDMPA/HCl-BSAO, a core-shell gold nanoparticles stabilized with the hydrophilic polymer (poly(3-dimethylammonium-1-propyne hydrochloride) and decorated with BSAO as described in the main text[45].…”

Theranostic nanomaterials applied to the cancer diagnostic and therapy and nanotoxicity studies

“…As a result of the formation of this bond, the peak at 2100 cm -1 (attributed to the N=C=S vibration in the Gd(III) chelate molecules) disappears and the peak at 1100 cm -1 (attributed to the C=S bound in the thiourea) appears after incorporation of the chelate into the silica layer. (202) In the next step, Gd-DOTA-SCN-doped APTES silica-coated Au colloid were incubated with Gd(NO 3 ) 3 to reload the Gd(III) ions, since the extremely conditions, such as high ammonia concentrations during the silica growth, can take out some ions from the chelates. The Gd(III) loaded within the SiO 2 layer before and after incubation with Gd(NO 3 ) 3 was examined by high resolution TEM (Figure 4.4).…”