Alternate adsorption of positively charged colloid-Au nanoparticles(nano-Au^(⊕))and negatively charged hemoglobin(Hb)on L-cysteine(L-cys)modified gold electrode resulted in the assembly of{Hb/nano-Au^(⊕)}n layer-by-layer films/L-cys modified gold electrode.The nano-Au^(⊕)was characterized by transmission electron micrograph(TEM)and microelectrophoresis.The modified electrode interface morphology was characterized by electrochemical impedance spectroscopy(EIS),atomic force mi-croscopy(AFM),cyclic voltammograms(CV)and chronoamperometry.Direct electron transfer between hemoglobin and gold electrodes was studied,and the apparent Michaelis-Menten constant(k_(m)^(app))of the modified electrode was evaluated to be 0.10 mmol·L^(-1).Moreover,the higher activity of proteins in the nano-Au^(⊕)films could be retained compared with the electropolymerization membrane,since the pro-teins in nano-Au^(⊕)films retained their near-native structure.Direct electron transfer between hemoglo-bin and electrode and electrochemically catalyzed reduction of hydrogen peroxide on a modified elec-trode was studied,and the linear range was from 2.1×10^(-8)to 1.2×10^(-3)mol·L^(-1)(r=0.994)with a detection limit of 1.1×10^(-8)mol·L^(-1)H_(2)O_(2).
YUAN RuoCAO ShuRuiCHAI YaQinGAO FengXianZHAO QingTANG MingYuTONG ZhongQiangXIE Yi
Electrochemical sensing of carcinoembryonic antigen(CEA)on a gold electrode modified by the se- quential incorporation of the mediator,thionine(Thi),and gold nanoparticles(nano-Au),through co- valent linkage and electrostatic interactions onto a self-assembled monolayer configuration is de- scribed in this paper.The enzyme,horseradish peroxidase(HRP),was employed to block the possible remaining active sites of the nano-Au monolayer,avoid the non-specific adsorption,instead of bovine serum albumin(BSA),and amplify the response of the antigen-antibody reaction.Electrochemical ex- periments indicated highly efficient electron transfer by the imbedded Thi mediator and adsorbed nano-Au.The HRP kept its activity after immobilization,and the studied electrode showed sensitive response to CEA and high stability during a long period of storage.The working range for the system was 2.5 to 80.0 ng/mL with a detection limit of 0.90 ng/mL.The model membrane system in this work is a potential biosensor for mimicking the other immunosensor and enzyme sensor.
