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Study of the Application and the Direct Electrochemistry of Hemoglobin Immobilized on Nanoparticles Modified Glassy Carbon Electrodes

Author: FanHong
Tutor: GuHaiYing
School: Nantong University
Course: Epidemiology and Biostatistics,
Keywords: Hemoglobin RBCs Nanoparticles Direct electron transfer Biosensor Electrogenerated chemiluminescence (ECL)
CLC: R115
Type: Master's thesis
Year: 2010
Downloads: 4
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Abstract


Objective (1) We presented a simple approach to synthesize a novelFe3O4@Pt core-shell nanocomposite, constructed a bionicfunctional interface based on hemoglobin (Hb) immobilizedon Fe3O4@Pt-chitosan (CS) film, denoted as“Hb-Fe3O4@Pt-CS”, and achieved the detection of H2O2.(2) A novel Fe3O4/APTES/CdS magnetic nanocomposite wassynthesized to study the direct electrochemistry and theelectrocatalysis of Hb immobilized on Fe3O4/APTES/CdS-CSmodified glassy carbon electrode (GCE).(3) The influence of Pt nanoparticles (NPs) on theelectrogenerated chemiluminescence (ECL) of CdSnanocrystals (NCs) with different fabrication approaches anddifferent amounts of Pt nanoparticles was studied. And abiosensor based on layer-by-layer assembly of Hb-Pt/CdSfilms was also constructed.(4) Red blood cells (RBCs) were immobilized on Au NPs-CSmodified GCE to study the transmembrane electron transfer ofHb inside RBCs.Methods (1) Fe3O4@Pt core-shell nanocomposite was synthesized byreduction of Pt4+onto the Fe3O4-core nanoparticles surfacesusing sodium dihydrogencitrate both as a reductant and astabilizer. Hb with positive charge and Fe3O4@Pt nanocomposite with negtive charge were adsorbed alternatelyonto CS modified GCE. The process was characterized bytransmission electron microscopy (TEM), ultraviolet-visiblespectroscopy (UV-vis), cyclic voltammetry (CV) andelectrochemical impendence spectroscopy (EIS).(2) Fe3O4-core nanoparticles were prepared by coprecipitationof an Fe(II) and Fe(III) chlorides in aqueous solutions withoutany surfactants. Then TGA-capped CdS nanoparticles wereconnected onto Fe3O4nanoparticles by3-aminopropyltriethoxysilane (APTES) to formFe3O4/APTES/CdS magnetic nanocomposite. The magneticnanocomposite was characterized by TEM, scanning electronmicroscopy (SEM) and energy dispersive X-ray spectroscopy(EDS), and used to immobilize Hb to construct a H2O2biosensor.(3) A brown stable Pt NPs were prepared without poisoningstabilizers, and then a series of Pt/CdS nanocompositesolutions containing an equal amount of CdS NCs anddifferent amount of Pt NPs were obtained. The influence of PtNPs on the ECL of CdS NCs with different fabricationapproaches and different amounts of Pt NPs were discussed.Meanwhile, a biosensor based on layer-by-layer assembly ofHb-Pt/CdS films was constructed.(4) RBCs were immobilized on Au NPs-CS modified GCE,and characterized by atomic force microscopy (AFM) andscanning electron microscope (SEM). The transmembraneelectron transfer and sensing behavior of Hb inside RBCswere systematically studied.Results (1) TEM indicated that Pt4+shell was successfully reduced onthe surface of Fe3O4. Hb immobilized on Fe3O4@Pt-CSmodified GCE remained its secondary structure and exhibitedgood catalytic activity towards H2O2.In pH7.0PBS,Hb-Fe3O4@Pt-CS modified GCE exhibited a couple ofwell-defined and quasi-reversible redox peaks. The anodic peak potential and cathodic peak potential were located at-0.38V and-0.32V respectively, the formal potential E0’wasabout-0.35V.(2) A novel Fe3O4/APTES/CdS magnetic nanocomposite wassuccessfully synthesized. Then Hb was immobilized on themagnetic nanocomposite to construct a H2O2biosensor. In pH7.0PBS, a reversible redox peaks of Hb was observed at-0.30V and-0.38V respectively. The formal potential E0’was about-0.34V. The linear range was from2.8×10-6to1.7×10-2mol/L,with a detection limit of4.0×10-7mol/L (S/N=3).(3) The influence of Pt NPs on the ECL of CdS NCs withdifferent fabrication approaches and different amounts of PtNPs were discussed. We found that an appropriateconcentration of Pt NPs can enhance the ECL intensity of CdSNCs, of which the excessive concentration showed aquenching effect. Besides, the {Hb-Pt/CdS}4multilayer filmsshowed good electrocatalytic activity to H2O2.(4) RBCs were successfully immobilized on Au NPs. Hbinside RBCs retained its original biological activity. In pH7.0PBS, a characteristic of the Hb-heme Fe(III)/Fe(II) redoxcouple was observed and the formal potential E0’was-0.35V.The electron transfer rate constant was0.89/s. The RBCs/AuNPs-CS modified GCE also displayed a sensitiveelectrochemical response to H2O2. The linear range was5.3×10-6to1.4×10-2mol/L with a detection limit of1.8×10-7mol/L (S/N=3).Conclusion:(1) Fe3O4@Pt core-shell nanoparticles were successfullysynthesized and Hb was immobilized on the nanocompositemodified GCE. A “Hb-Fe3O4@Pt-CS” bionic functionalinterface was then constructed, while direct electron transferof Hb was successfully achieved.(2) Hb was successfully immobilized on Fe3O4/APTES/CdSmagnetic nanocomposites, and the electron transfer of Hb wasachieved rapidly. The obtained modified GCE also displayed a good catalytic performance to H2O2.(3) We found that an appropriate concentration of Pt NPscould enhance the ECL intensity of CdS NCs, and theobtained {Hb-Pt0.004/CdS1}4-CS modified GCE showed goodelectrocatalytic activity to H2O2.(4) Transmembrane direct electron transfer of Hb inside RBCsimmobilized on Au NPs was successfully achieved, and aH2O2biosensor based on RBCs/Au NPs-CS modified GCEwas then constructed.

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