General Preparation of Heme Protein Functional Fe₃O₄@Au‐Nps Magnetic Nanocomposite for Sensitive Detection of Hydrogen Peroxide

Abstract

1IntroductionDirect electron transfer (DET) of heme protein has been aresearch focus becauseitnot only establishes adesirable model for fundamental study of proteins in biological systems,but also plays asignificant role in fabricating mediator-free bioelectrochemicald evices,s uch as the third generation biosensors [1],b iofuel cells [2],e nzymatic bioreactors [3] and biomedical devices [4].H emoglobin (Hb) is ak ind of utmost important heme protein in red cells.I t has aq uaternary structurec ontaining four polypeptide chains (globin chains)a nd one heme group bound to each globin chain [5,6].D ue to its known structurea nd commercial availability,H bi sc onsidered as an ideal model molecule for heme protein study.U nfortunately, DET between Hb and electrodes urface is generally difficult becauset he redox-active center is embedded in its protein shelld eeply [7] and enzyme usually losesi ts bioactivity on the bare electrode [8].T of acilitate DET and retain the bioactivity of immobilizede nzyme,b iocompatible materials have been used to immobilize Hb,s uch as surfactants [9],b iopolymers [10],h ydrogels [11],p olyelectrolytes [12] and nano-materials [13].Considerable attentions have been paid to nano-materials due to their remarkable advantages includingh igh conductivity,h igh catalytic efficiency and excellentb iocompatibility.T hey will present favorable microenvironment to keep the activity of immobilizedp roteins and consequently to fabricate eligible biosensors.F or exam-ple,A un anoparticles (Au-NPs) can offer an attractive platformf or enzyme immobilization not only because of its excellent conductivitya nd biocompatibility but also because of its facile and robust interaction with various biomolecules containing thiol and disulfideg roups [14,15].B esides,m agnetic nanoparticles are also strong candidates in al argev ariety of applications. They are ideal bimolecular immobilizingc arriers and they enable quick and efficient isolation or extraction of at arget molecule or substance by an externalm agnetic field [16].T he most widely studied superparamagneticn anoparticles are magnetite (Fe 3 O 4 ). However, naked Fe 3 O 4 nanoparticles are very sensitive to oxidation and pronet oa ggregate [17].O ne of the maina pproaches to overcome these limitations is to protect naked magnetic NPs with metals [18].Abstract:S table magneticn anocomposite of gold nanoparticles (Au-NPs) decoratingF e 3 O 4 core was successfully synthesized by the linker of Boc-L-cysteine.T ransmission electron microscope (TEM), energy dispersive X-ray spectroscopy (EDX)a nd cyclic voltammograms (CV) were performedt oc haracterize the as-prepared Fe 3 O 4 @Au-Nps.T he results indicated that Au-Nps dispersed homogeneously around Fe 3 O 4 with the ratio of Au to Fe 3 O 4 nanoparticles as 5-10/1 andt he apparente lectrochemical area as 0.121 cm 2 .A fters elf-assembly of hemoglobin (Hb)o nF e 3 O 4 @Au-Npsb ye lectrostatic interaction,ahydrogen peroxide biosensor was developed. TheF e 3 O 4 @Au-Nps/Hbm odified GCE exhibited fast direct ele...

Detection Technologies for Reactive Oxygen Species: Fluorescence and Electrochemical Methods and Their Applications

“…Myoglobin and hemin are usually immobilized on AuNPs due to their good conductivity, excellent biocompatibility, and the ability to easily interact with proteins via the thiol groups and disulfide bonds, leading to a simple biosensor preparation. For example, in 2016, Xu et al [ 150 ] took advantage of AuNPs to improve the sensor efficiency for the detection of H 2 O 2 using Hb as the biological redox protein. To do so, they immobilized Hb on a magnetic Fe 3 O 4 /AuNPs nanocomposite.…”

“…Porous materials have also been used for immobilization of redox proteins. Various conductive complex materials, such as Ag-Au [ 162 ], Fe 3 O 4 -Au [ 150 ], polyethylenimine (PEI)-Au [ 163 ], and GO-chitosan-Au [ 159 , 160 , 161 , 164 ], have been reported in the literature. As previously mentioned, one of the main reasons for integrating redox proteins with conductive substrates with large surface areas is to improve the amount and stability of bound redox proteins on the electrode surface.…”

“…Myoglobin and hemin are usually immobilized on AuNPs due to their good conductivity, excellent biocompatibility, and the ability to easily interact with proteins via the thiol groups and disulfide bonds, leading to a simple biosensor preparation. For example, in 2016, Xu et al [150] As previously stated, graphene oxide (GO) and carbon nanotubes (CNTs) have also been frequently combined with biological redox proteins. For example, hemin was immobilized on either GO or CNTs to reduce the challenges associated with its use for the detection of H 2 O 2 [151][152][153].…”

Preparation and Properties of a Low Fouling Magnetic Nanoparticle and Its Application to the HPV Genotypes Assay in Whole Serum

“…One case is that Han et al constructed an electrochemical biosensor based on a Fe 3 O 4 @Au@GSH magnetic sensing film to detect estradiol . Another case is that Xu et al sensitively detected hydrogen peroxide by heme protein functional Fe 3 O 4 @Au nanoparticles (NPs) …”

Construção e otimização de uma plataforma sensorial eletroquímica a base de nano-metalopolimero poli[Ni(<i>Salpn</i>)]

“…Atualmente as determinações de H 2 O 2 em alimentos e bebidas industrializadas são realizadas por espectroscopia e fluorescência . Recentemente muitos estudados relatam a aplicação de proteínas Heme funcionais na construção de eletrodos para determinação de H 2 O 2[32,33], apresentando alta seletividade.Entretanto, o uso de enzimas, requer cuidados especiais no âmbito que tange alguns dos principais limitadores da estabilidade enzimática, como por exemplo, a estocagem, necessidade de pré-tratamento e complexos procedimentos experimentais, além de um custo elevado quando comparado aos eletrodos que não envolvem tais processos.…”