Construction of an improved amperometric acrylamide biosensor based on hemoglobin immobilized onto carboxylated multi-walled carbon nanotubes/iron oxide nanoparticles/chitosan composite film

A method is described for construction of an improved amperometric acrylamide biosensor based on covalent immobilization of hemoglobin (Hb) onto nanocomposite of carboxylated multi-walled carbon nanotubes (cMWCNT) and iron oxide nanoparticles (Fe₃O₄NPs) electrodeposited onto Au electrode through chitosan (CHIT) film. The Hb/cMWCNT-Fe₃O₄NP/CHIT/Au electrode was characterized by scanning electron microscopy, Fourier transform infra-red spectroscopy, electrochemical impedance spectroscopy, and differential pulse voltammetry at different stages of its construction. The biosensor was based on interaction between acrylamide and Hb, which led to decrease in the electroactivity of Hb, i.e., current generated during its reversible conversion [Fe(II)/Fe(III)]. The biosensor showed optimum response within 8 s at pH 5.0 and 30 °C. The linear working range for acrylamide was 3–90 nM, with a detection limit of 0.02 nM and sensitivity of 36.9 μA/nM/cm². The biosensor was evaluated and employed for determination of acrylamide in potato crisps.     

A magnetic molecular imprinting-based fluorescence probe for sensitive and selective detection of 2,4-D herbicide

A new magnetic molecular imprinting-based turn-on fluorescence probe (Fe₃O₄NPs@SiO₂@NBD@MIPs) has been synthesized via a facile sol–gel polymerization for the detection of 2,4-dichlorophenoxyacetic acid (2,4-D). Based on the photoinduced electron transfer (PET) of nitrobenzoxadiazole (NBD), 2,4-D can be recognized by enhancement of NBD fluorescence. With the presence of Fe₃O₄ in the core of the probe, this sensor can also be reused many times using magnetic aggregation methods. After the addition of various concentrations of 2,4-D, the fluorescence peak at 530 nm (excitation of 468 nm) increased linearly ranging from 0.1 to 3 μM with a detection limit of 0.023 μM. This sensing system is believed to be available for detecting 2,4-D in real samples, with high recovery rates ranging from 94% to 108% for three spike levels of 2,4-D with precisions below 5%.     

In vitro spectroscopic investigation of groove binding interaction of Fe3O4@CaAl-LDH@L-Dopa with calf thymus DNA

Abstract

The principal goal of this study is to evaluate the interaction of Fe₃O₄@CaAl-LDH@L-Dopa and Fe₃O₄@CaAl-LDH nanoparticles with calf thymus DNA. The magnetic nanoparticles were previously prepared by a chemical co-precipitation method, and the surface of the Fe₃O₄ nanoparticles was coated with CaAl layered double hydroxides. The antiparkinsonian drug “L-Dopa” was carried by this core–shell nanostructure to achieve the drug delivery system with suitable properties for biological applications. Also, the interaction of Fe₃O₄@CaAl-LDH@L-Dopa and Fe₃O₄@CaAl-LDH nanoparticles with CT-DNA was studied using, UV–Visible spectroscopy, viscosity, circular dichroism (CD), and fluorescence spectroscopy techniques. The results of investigations demonstrated that Fe₃O₄@CaAl-LDH@L-Dopa and Fe₃O₄@CaAl-LDH nanoparticles have interacted via minor groove binding and intercalated to CT-DNA, respectively.     

Magnetic nanocomposite of anti-human IgG/COOH-multiwalled carbon nanotubes/Fe₃O₄ as a platform for electrochemical immunoassay

An electrochemical immunosensing method was developed based on a magnetic nanocomposite. The multiwalled carbon nanotubes (MWCNTs) were treated with nitric acid to produce carboxyl groups at the open ends. Then, Fe₃O₄ nanoparticles were deposited on COOH–MWCNTs by chemical coprecipitation of Fe²⁺ and Fe³⁺ salts in an alkaline solution. Goat anti-human IgG (anti-hIgG) was covalently attached to magnetic nanocomposite through amide bond formation between the carboxylic groups of MWCNTs and the amine groups of anti-hIgG. The prepared bio-nanocomposite was used for electrochemical sensing of human tetanus IgG (hIgG) as a model antigen. The anti-hIgG magnetic nanocomposite was fixed on the surface of a gold plate electrode using a permanent magnet. The hIgG was detected using horseradish peroxidase (HRP)-conjugated anti-hIgG in a sandwich model. Electrochemical detection of hIgG was carried out in the presence of H₂O₂ and KI as substrates of HRP. Using this method, hIgG was detected in a concentration range from 30 to 1000 ng ml^?1 with a correlation coefficient of 0.998 and a detection limit of 25 ng ml^?1 (signal/noise = 3). The designed immunosensor was stable for 1 month.     

Effect of nanoheat stimulation mediated by magnetic nanocomposite hydrogel on the osteogenic differentiation of mesenchymal stem cells

Hyperthermia has been considered as a promising healing treatment in bone regeneration. We designed a tissue engineering hydrogel based on magnetic nanoparticles to explore the characteristics of hyperthermia for osteogenic regeneration. This nanocomposite hydrogel was successfully fabricated by incorporating magnetic Fe₃O₄ nanoparticles into chitosan/polyethylene glycol (PEG) hydrogel, which showed excellent biocompatibility and were able to easily achieve increasing temperatures under an alternative magnetic field (AMF). With uniformly dispersed nanoparticles, the composite hydrogel resulted in high viability of mesenchymal stem cells (MSCs), and the elevated temperature contributed to the highest osteogenic differentiation ability compared with direct heat treatment applied under equal temperatures. Therefore, the nanoheat stimulation method using the magnetic nanocomposite hydrogel under an AMF may be considered as an alternative candidate in bone tissue engineering regenerative applications.     

Biodesulfurization of Dibenzothiophene by Microbial Cells Coated with Magnetite Nanoparticles

Microbial cells of Pseudomonas delafieldii were coated with magnetic Fe₃O₄ nanoparticles and then immobilized by external application of a magnetic field. Magnetic Fe₃O₄ nanoparticles were synthesized by a coprecipitation method followed by modification with ammonium oleate. The surface-modified Fe₃O₄ nanoparticles were monodispersed in an aqueous solution and did not precipitate in over 18 months. Using transmission electron microscopy (TEM), the average size of the magnetic particles was found to be in the range from 10 to 15 nm. TEM cross section analysis of the cells showed further that the Fe₃O₄ nanoparticles were for the most part strongly absorbed by the surfaces of the cells and coated the cells. The coated cells had distinct superparamagnetic properties. The magnetization (δ_s) was 8.39 emu · g⁻¹. The coated cells not only had the same desulfurizing activity as free cells but could also be reused more than five times. Compared to cells immobilized on Celite, the cells coated with Fe₃O₄ nanoparticles had greater desulfurizing activity and operational stability.     

Electrospinning of Cross-Linked Magnetic Chitosan Nanofibers for Protein Release

A poly(vinylalcohol) (PVA) electrospun/magnetic/chitosan nanocomposite fibrous cross-linked network was fabricated using in situ cross-linking electrospinning technique and used for bovine serum albumin (BSA) loading and release applications. Sodium tripolyphosphate (TPP) and glutaraldehyde (GA) were used as cross-linkers which modified magnetic-Fe₃O₄ chitosan as Fe₃O_4/CS/TPP and Fe₃O_4/CS/GA, respectively. BSA was used as a model protein drugs which was encapsulated to form Fe₃O₄/CS/TPP/BSA and Fe₃O₄/CS/GA/BSA nanoparticles. The composites were electrospun with PVA to form nanofibers. Nanofibers were characterized by field emission scanning electron microscopy (FESEM) and Fourier transform infrared spectroscopy (FTIR). The characterization results suggest that Fe₃O₄ nanoparticles with average size of 45 nm were successfully bound on the surface of chitosan. The cross-linked nanofibers were found to contain uniformly dispersed Fe₃O₄ nanoparticles. The size and morphology of the nanofibers network was controlled by varying the cross-linker type. FTIR data show that these two polymers have intermolecular interactions. The sample with TPP cross-linker showed an enhancement of the controlled release properties of BSA during 30-h experimental investigation.

Graphical Abstract

Open in a separate windowᅟKEY WORDS: cross-linker, electrospun, magnetite, mano-composite, protein loading     

Conjugation of nattokinase and lumbrukinase with magnetic nanoparticles for the assay of their thrombolytic activities

Two important thrombolytic enzymes, nattokinase (NK) and lumbrukinase (LK), were immobilized onto fine magnetic Fe₃O₄ nanoparticles using 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide (EDC) as the coupling reagent, and their thrombolytic activities were studied. The Fe₃O₄ nanoparticles and NK- and LK-conjugated magnetic nanoparticles were characterized by transmission electron microscopy, Fourier transform infrared spectrophotometry, vibrating sample magnetometry, X-ray diffraction, and UV–vis absorption spectroscopy. Dual kinetic absorbance measurements at 405 and 630 nm were employed to measure their thrombolytic activity. Analysis of protein amount showed that the optimum conditions for NK and LK binding to nanoparticles were respectively at a mass ratio of 2:1:1, 2:1:2 (magnetic nanoparticles:protein:EDC), and pH 6.00. Thrombolytic activity assay showed that the best thrombolytic activity could reach 91.89% for NK–nanoparticle conjugates and 207.74% for LK–nanoparticle conjugates, which are much higher than the pure enzymes (NK, 82.86%; LK, 106.57%).     

DNA binding and cytotoxicity studies of magnetic nanofluid containing antiviral drug oseltamivir

In this work, the possibility of preparing a nanoparticle with improved treatment properties was investigated. In this regard, synthesis, characterization, in vitro cytotoxicity and DNA binding of Fe₃O₄@oleate/oseltamivir magnetic nanoparticles (MNPs) were investigated. Fe₃O₄ nanoparticles were synthesized via chemical co-precipitation and coated by oleate bilayers. Then, Fe₃O₄@OA MNPs were functionalized with an antiviral drug (oseltamivir), for better biological applications. The MNPs were subsequently characterized by zeta sizer and Zeta potential measurements, Fourier transform infrared (FT-IR) spectroscopy, vibrating sample magnetometer (VSM) and transmission electron microscopy (TEM) analyses. The TEM image demonstrated that average sizes of Fe₃O₄@OA/oseltamivir MNPs were about 8?nm. The in vitro cytotoxicity of Fe₃O₄@OA/oseltamivir MNPs was studied against cancer cell lines (MCF-7 and MDA-MB-231) and compared with oseltamivir drug. The results illustrated that Fe₃O₄@OA/oseltamivir magnetic nanoparticles have better antiproliferative effects on the mentioned cell lines as compared with oseltamivir. Also, in vitro DNA binding studies were done by UV–Vis, circular dichroism, and Fluorescence spectroscopy. The results indicated that Fe₃O₄@OA/oseltamivir MNPs bound to DNA via groove binding. Moreover, this magnetic nanofluid has potential for magnetic hyperthermia therapy due to magnetic core of its nanoparticles.

Communicated by Ramaswamy H. Sarma     

Novel and efficient method for immobilization and stabilization of β-d-galactosidase by covalent attachment onto magnetic Fe3O4–chitosan nanoparticles

A novel and efficient immobilization of β-d-galactosidase from Aspergillus oryzae has been developed by using magnetic Fe₃O₄–chitosan (Fe₃O₄–CS) nanoparticles as support. The magnetic Fe₃O₄–CS nanoparticles were prepared by electrostatic adsorption of chitosan onto the surface of Fe₃O₄ nanoparticles made through co-precipitation of Fe²⁺ and Fe³⁺. The resultant material was characterized by transmission electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, vibrating sample magnetometry and thermogravimetric analysis. β-d-Galactosidase was covalently immobilized onto the nanocomposites using glutaraldehyde as activating agent. The immobilization process was optimized by examining immobilized time, cross-linking time, enzyme concentration, glutaraldehyde concentration, the initial pH values of glutaraldehyde and the enzyme solution. As a result, the immobilized enzyme presented a higher storage, pH and thermal stability than the soluble enzyme. Galactooligosaccharide was formed with lactose as substrate by using the immobilized enzyme as biocatalyst, and a maximum yield of 15.5% (w/v) was achieved when about 50% lactose was hydrolyzed. Hence, the magnetic Fe₃O₄–chitosan nanoparticles are proved to be an effective support for the immobilization of β-d-galactosidase.     

Magnetic Clustering Effect during the Association of Biofunctionalized Magnetic Nanoparticles with Biomarkers

We report herein an investigation into dynamic magnetic clustering that occurs during immunoassays as biofunctionalized magnetic nanoparticles (BMNs) become associated with biotargets. We measure the dynamic effective relaxation time τ _eff(t) and use scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to investigate the C-reactive protein (CRP) as it associates with the BMN Fe₃O₄-antiCRP to form the magnetic cluster Fe₃O₄-antiCRP-CRP. The results indicate that τ _eff(t) increases with increasing association time. In addition, the ration Δτ _eff/τ ₀ as a function of CRP concentration follows a characteristic logistic function, which provides a basis for estimating the quantity of biomolecules with a detection sensitivity close to 0.1 ppm. After the association, SEM and TEM images show that CRP and Fe₃O₄-antiCRP conjugate to form Fe₃O₄-antiCRP-CRP clusters hundreds of nanometers in size. The SEM and TEM images provide direct evidence of the formation of magnetic clustering.     

A novel electroconductive interface based on Fe3O4 magnetic nanoparticle and cysteamine functionalized AuNPs: Preparation and application as signal amplification element to minoring of antigen‐antibody immunocomplex and biosensing of prostate cancer

In this study, a novel electroconductive interface was prepared based on Fe₃O₄ magnetic nanoparticle and cysteamine functionalized gold nanoparticle. The engineered interface was used as signal amplification substrate in the electrochemical analysis of antibody‐antigen binding. For this purpose, biotinilated‐anti‐prostate‐specific antigen (PSA) antibody was bioconjugated with iron oxide magnetic nanoparticles (Fe₃O₄) and drop‐casted on the surface of glassy carbon electrode (GCE). Also, secondary antibody (HRP‐Ab2) encapsulated on gold nanoparticles caped by cysteamine was immobilized on the surface of GCE modified electrode. A transmission electron microscopy images shows that a sandwich immunoreaction was done and binding of Ab1 and Ab2 performed successfully. Various parameters of immunoassay, including the loading of magnetic nanoparticles, the amount of gold nanoparticle conjugate, and the immunoreaction time, were optimized. The detection limit of 0.001 μg. L^?1 of PSA was obtained under optimum experimental conditions. It is found that such magneto‐bioassay could be readily used for simultaneous parallel detection of multiple proteins by using multiple inorganic metal nanoparticle tracers and are expected to open new opportunities for early stage diagnosis of cancer in near future.     

3D Porous γ‐Fe2O3@C Nanocomposite as High‐Performance Anode Material of Na‐Ion Batteries

A simple and template‐free method for preparing three‐dimensional (3D) porous γ‐Fe₂O₃@C nanocomposite is reported using an aerosol spray pyrolysis technology. The nanocomposite contains inner‐connected nanochannels and γ‐Fe₂O₃ nanoparticles (5 nm) uniformly embedded in a porous carbon matrix. The size of γ‐Fe₂O₃ nanograins and carbon content can be controlled by the concentration of the precursor solution. The unique structure of the 3D porous γ‐Fe₂O₃@C nanocomposite offers a synergistic effect to alleviate stress, accommodate large volume change, prevent nanoparticles aggregation, and facilitate the transfer of electrons and electrolyte during prolonged cycling. Consequently, the nanocomposite shows high‐rate capability and long‐term cyclability when applied as an anode material for Na‐ion batteries (SIBs). Due to the simple one‐pot synthesis technique and high electrochemical performance, 3D porous γ‐Fe₂O₃@C nanocomposites have a great potential as anode materials for rechargeable SIBs.     

An electrochemical immunosensor for digoxin using core–shell gold coated magnetic nanoparticles as labels

A simple, sensitive, and low-cost immunosensor was designed for the detection of digoxin through core–shell gold coated magnetic nanoparticles (Fe₃O₄-Au-NPs) as an electrochemical label. Having had such a large potential for a variety of applications, Fe₃O₄-Au-NPs have attracted a considerable attention and are actively investigated recently. Digoxin is a cardiac glycoside which, at high level, can indicate an increased risk of toxicity. This new competitive electrochemical immunosensor was developed based on antigen–antibody reaction employing antigen (Ag) labeled Fe₃O₄-Au-NPs and PVA modified screen-printed carbon electrode surface in order to detect the serum digoxin. The structures of Fe₃O₄-Au-NPs were studied by transmission electron microscopy, X-ray diffraction and Fourier transformed infrared spectroscopy. Cyclic voltammetry and differential pulse voltammetry (DPV) were employed to determine the physicochemical and electrochemical properties of immunosensor. DPV was employed for quantitative detection of digoxin in biological samples. The developed immunosensor was capable to detect digoxin in the range from 0.5 to 5 ng mL^?1, with a detection limit as low as 0.05 ng mL^?1. The proposed method represented acceptable reproducibility, stability, and reliability for the rapid detection of digoxin in serum samples.     

Preparation and characterization of magnetic gold nanoparticles to be used as doxorubicin nanocarriers

Magnetic targeted drug delivery (MTD), using magnetic gold nanoparticles (Fe₃O₄@Au NPs) conjugated with an anti-cancer drug is a promise modality for cancer treatment. In this study, Fe₃O₄@Au NPs were prepared and functionalized with thiol-terminated polyethylene glycol (PEG), then loaded with anti-cancer drug doxorubicin (DOX). The physical properties of the prepared NPs were characterized using different techniques. Transmission electron microscopy (TEM) revealed the mono dispersed nature of Fe₃O₄@Au NPs with an average size of 20 nm which was confirmed using Dynamic light scattering (DLS) measurements. Zeta potential measurements along with UV–VIS spectroscopy demonstrated surface DOX loading on Fe₃O₄@Au NPs. Energy Dispersive X-ray Spectroscopy (EDX) assured the existence of both iron and gold elements in the prepared NPs. The paramagnetic properties of the prepared NPs were assessed by vibrating sample magnetometer (VSM). The maximum DOX-loading capacity was 100 μg DOX/mg of Fe₃O₄@Au NPs. It was found that DOX released more readily at acidic pH. In vitro studies on MCF-7 cell line elucidated that DOX loaded Fe₃O₄@Au NPs (Fe₃O₄@Au-PEG-DOX) have more potent therapeutic effect than free DOX. Knowledge gained in this study may open the door to pursue Fe₃O₄@Au NPs as a viable nanocarriers for different molecules delivery in many diagnostic and therapeutic applications.     

Magnetic core/shell Fe3O4/Au nanoparticles for studies of quinolones binding to protein by fluorescence spectroscopy

Magnetic core/shell Fe₃O₄/Au nanoparticles were used in the determination of drug binding to bovine serum albumin (BSA) using a fluorescence spectroscopic method. The binding constants and number of binding sites for protein with drugs were calculated using the Scatchard equation. Because of their superparamagnetic and biocompatible characteristics, magnetic core/shell Fe₃O₄/Au nanoparticles served as carrier proteins for fixing proteins. After binding of the protein to a drug, the magnetic core/shell Fe₃O₄/Au nanoparticles–protein–drug complex was separated from the free drug using an applied magnetic field. The free drug concentration was obtained directly by fluorescence spectrometry and the proteins did not influence the drug determination. So, the achieved number of binding sites should be reliable. The binding constant and site number for ciprofloxacin (CPFX) binding to BSA were 2.055 × 10⁵ L/mol and 31.7, and the corresponding values for norfloxacin (NOR) binding to BSA were 1.383 × 10⁵ L/mol and 38.8. Based on the achieved results, a suitable method was proposed for the determination of binding constants and the site number for molecular interactions. The method was especially suitable for studies on the interactions of serum albumin with the active ingredients of Chinese medicine. Copyright © 2015 John Wiley & Sons, Ltd.     

Conjugation of α-chymotrypsin on a polymeric hydrophilic nanolayer covering magnetic nanoparticles

Magnetic nanoparticles, covered by a polymeric hydrophilic nanolayer containing reactive amino groups, were obtained via Hoffman degradation of the polyacrylamide-coated Fe₃O₄ nanoparticles synthesized by photochemical in situ polymerization, and then conjugated the model enzyme––α-chymotrypsin (CT) by use of EDC· HCl and NHS at room temperatures. The mechanism of photochemical in situ polymerization was briefly proposed in this paper. Superparamagnetic properties were retained for Fe₃O₄ after enzyme immobilization while slightly reducing the value of saturation magnetization. Crystalline structure of Fe₃O₄ after CT immobilization was consistent with that of the freshly prepared Fe₃O₄ by X-ray diffraction (XRD) analysis. The binding capacity was 69 and 61 mg enzyme/g nanogel determined by thermogravimetric (TG) analysis and by standard BCA protein assay, respectively. Specific activity of the immobilized CT was 0.93 U/(mg min), only 59.3% as that of free CT. Thermal stability of CT was improved after being bound to the amine-functionalized magnetic nanogel.     

Optimization of the covalent coupling and ionic adsorption of magnetic nanoparticles on Flavobacterium ATCC 27551 using the Taguchi method

Abstract

Flavobacterium ATCC 27551 was used as a model system for the preparation of magnetic biocatalysts. The magnetic modification was carried out by covalently binding carboxylate- and amino-modified magnetic nanoparticles onto cells. Magnetic Fe₃O₄ nanoparticles were also used for ionic adsorption on the cell surface. Magnetically modified cells were concentrated using a magnet and exhibited organophosphate hydrolyzing activity. The Taguchi method was used to optimize the binding of the magnetic nanoparticles on the cell surface. SEM image analyses demonstrated good linkage of the magnetic nanoparticles over the Flavobacterium ATCC 27551 cell surface. Under optimal conditions, the magnetic cells displayed specific activity ratios of 93%, 89% and 95%, compared with untreated cells, after the covalent coupling with carboxylate- and amino-modified magnetic nanoparticles and the ionic adsorption of magnetic Fe₃O₄ nanoparticles, respectively.     

The function of chitosan/agarose biopolymer on Fe2O3 nanoparticles and evaluation of their effects on MCF-7 breast cancer cell line and expression of BCL2 and BAX genes

In recent decades, magnetic nanoparticles modified with biocompatible polymers have been recognized as a suitable tool for treating breast cancer. The aim of this research was to evaluate the function of chitosan/agarose-functionalized Fe₂O₃ nanoparticles on the MCF-7 breast cancer cell line and the expression of BCL2 and BAX genes. Free Fe₂O₃ nanoparticles were prepared by hydrothermal method. FTIR, XRD, SEM, DLS, VSM, and zeta potential analyses determined the size and morphological characteristics of the synthesized nanoparticles. The effect of Fe₂O₃ free nanoparticles and formulated Fe₂O₃ nanoparticles on induction of apoptosis was studied by double-dye Annexin V-FITC and PI. Also, the gene expression results using the PCR method displayed that Fe₂O₃ formulated nanoparticles induced BAX apoptosis by increasing the anti-apoptotic gene expression and decreasing the expression of pro-apoptotic gene BCL2, so the cell progresses to planned cell death. In addition, the results showed that the BAX/BCL2 ratio decreased significantly after treatment of MCF-7 cells with free Fe₂O₃ nanoparticles, and the BAX/BCL2 ratio for Fe₂O₃ formulated nanoparticles increased significantly. Also, to evaluate cell migration, the scratch test was performed, which showed a decrease in motility of MCF-7 cancer cells treated with Fe₂O₃ nanoparticles formulated with chitosan/agarose at concentrations of 10, 50, 100, and 200 μg/ml.