Silica sol-gel immobilized amperometric biosensor for hydrogen peroxide

Li, Juan; Tan, Swee Ngin; Ge, Honghua

doi:10.1016/s0003-2670(96)00337-6

Cited by 302 publications

(137 citation statements)

References 21 publications

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“…Thus, the apparent Michaelis-Menten constant (K M ), which indicates the kinetics of enzyme-substrate reaction, was calculated according to the electrochemical version of the LineweaverBurk equation. 50,51 The reduction peak current increased with the concentration of H 2 O 2 enhanced and at higher concentration, a response plateau was observed (data not shown). 52 1 / I ss = 1 / I max + K M / I max C Where I ss is the steady-state current after the addition of substrate, C is the bulk concentration of the substrate, I max is the maximum current measured at the saturated substrate conditions.…”

Section: Spr Studies Of the Layers Formation On Gold Surfacementioning

confidence: 94%

mentioning

confidence: 94%

“…This K M value is intermediary than some reported values. [53][54][55][56] As is well known, the smaller K M shows the higher catalytic ability and it also indicates a larger affinity of the enzyme to the substrate. These results indicated that the peroxidase activity of entrapped HRP was maintained.…”

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confidence: 99%

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Kinetic studies of HRP adsorption on ds-DNA immobilized on gold electrode surface by EIS and SPR

Ferreira

Kubota

2010

J. Braz. Chem. Soc.

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Este trabalho descreve a construção de um filme de multicomponentes via método layer-bylayer (LbL) e a análise cinética da interação da enzima peroxidase de raiz forte (HRP) sobre a camada de ds-DNA de timo de bezerro formada numa superfície de ouro modificada. As técnicas de ressonância de plásmon de superfície (SPR) e a espectroscopia de impedância eletroquímica (EIE) foram usadas para caracterizar a construção do filme sobre a superfície de ouro. A ressonância de plásmon de superfície forneceu informações essenciais para o estudo e caracterização da interação proteína e ácido nucléico. Este método não necessita de marcador e monitora as interações em tempo real. Os estudos cinéticos obtidos por SPR da formação do filme de HRP sobre o ds-DNA apresentaram valores de 24,7 L mol -1 s -1 e 1,2×10 -3 s -1 para k a e k d , respectivamente. A energia livre de Gibbs obtida para o sistema foi de -23,1 kJ mol -1. Os resultados obtidos mostram que a interação das moléculas da enzima sobre o ds-DNA é cinética e termodinamicamente favoráveis e a interação entre as camadas ocorre principalmente pela atração de cargas opostas. This paper describes the construction of a multicomponent film via layer-by-layer (LbL) method and the kinetic analysis of the interaction between horseradish peroxidase (HRP) enzyme on calf thymus double-stranded DNA layer on a modified gold surface. Surface plasmon resonance (SPR) and electrochemical impedance spectroscopy (EIS) are used to characterize the successful construction of the film on the gold surface. Surface plasmon resonance provided essential information for the study and characterization of protein and nucleic acid interaction and this method is label-free and monitors the interactions in real time. The kinetic studies determined by SPR of the horseradish peroxidase film formation on ds-DNA layer showed values of 24.7 L mol -1 s -1 and 1.2×10 -3 s -1 for k a and k d , respectively. The Gibbs free energy obtained for the system was -23.1 kJ mol -1. The results obtained show that the interaction of the enzymes molecules on ds-DNA is kinetically and thermodynamically favourable and the interaction among the layers probably occurs mainly by attraction of opposite charges.Keywords: SPR, EIS, kinetic adsorption, DNA, HRP, protein-DNA interaction IntroductionThe interaction of nucleic acids with protein has been a primary area of interest for therapeutic development and in the study of cellular processes.1-3 Development of devices based on this interaction presents application in the investigations of the toxicity pathways of nucleic acid. Recently, electrochemical sensors have been developed for the rapid detection of DNA damage. [4][5][6][7] Such devices are being successfully used in food industry, environmental fields and clinical chemistry. Biosensors for such analysis can be prepared with multicomponent films of polyions and other charged materials and to achieve high specificity, high sensitivity, rapid response and flexibility of use, it is clear that the research continues to focus ...

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Section: Spr Studies Of the Layers Formation On Gold Surfacementioning

confidence: 94%

mentioning

confidence: 94%

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Kinetic studies of HRP adsorption on ds-DNA immobilized on gold electrode surface by EIS and SPR

Ferreira

Kubota

2010

J. Braz. Chem. Soc.

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“…The relationship between catalytic current and the concentration of glucose was considered based on the electrochemical version of Michaelis-Menten plot. The enzyme kinetic parameters of apparent Michaelis-Menten constant (K ap m ) and maximum current (I max ) measured under substrate saturated concentration can be calculated based on Lineweaver-Burk (5) [24] where I ss stands for steady state current. From the slope and intercept of the plot, the values of K ap m and I max for electrode A were estimated to be 18 μM and 2.7 μA, respectively.…”

Section: Amperometric Detection Of Glucosementioning

confidence: 99%

A Biocompatible Nanocomposite for Glucose Sensing

Rahimi

Ghourchian

Rafiee‐Pour

et al. 2011

International Journal of Electrochemistry

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A nanocomposite containing amine functionalized multiwalled carbon nanotubes and a room temperature ionic liquid (1-butyl-3-methylimidazolium tetrafluoroborate) was prepared and applied for glucose oxidase (GOx) immobilization on glassy carbon electrode. The proposed nanocomposite provided a favorable microenvironment to preserve the bioactivity of GOx. It could also effectively facilitate the enzyme direct electron transfer to the electrode. This brought about a remarkable improvement in the sensitivity of the glucose biosensor. Under the optimum experimental conditions, the formal potential of GOx was about −467.5 mV. Moreover, the sensitivity and response time of the biosensor toward glucose were 1277 μA mM −1 cm −2 and 6 s, respectively. The biosensor provided a linear dynamic response between 0.1 and 43 μM with a very low detection limit of 63 nM. Also, the values for apparent Michaelis-Menten constant and maximum current were obtained as 18 μM and 2.7 μA, respectively.

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“…41,42 The second one was sol-gel derivated ceramic carbon composite materials which was extensively used in amperometric biosensor. 43,44 The third one was sol-gel derivated inorganic-organic hybrid composite material, which integrated the merits of sol-gel technique and hydrogel entrapment. 45 Organic modification of sol-gel derivated matrices is used to reduce the degree of cross-link, improve film adhesion to its supports, or introduce reactive functional groups that can be subsequently used for anchoring of molecular recognition species on pre-prepared xerogels.…”

Section: Introductionmentioning

confidence: 99%

Strategies for developing NADH detector based on meldola blue in different immobilization methods: a comparative study

Mariotti¹,

Riccardi²,

Fertonani³

et al. 2006

J. Braz. Chem. Soc.

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Foi realizado um estudo comparativo entre duas metodologias de imobilização do Azul de Meldola (MB), utilizado para diminuir o potencial de oxidação do NADH. O estudo foi realizado no eletrodo de carbono vítreo através da eletropolimerização do pirrol e MB, empregando a técnica de voltametria cíclica e avaliando-se o efeito do número de ciclos para definir a espessura e a estabilidade do filme. Com 25 ciclos foi obtido o melhor resultado. Foi também investigada a modificação do eletrodo de grafite com filme formado pelo sistema sol-gel PPO 300 ou PPO 4000 pelo método de recobrimento por mergulho, através de dois métodos: oclusão e adsorção. O melhor método foi o de adsorção da MB durante 10 min, seguida de depósito do filme PPO 300, favorecido pelo tempo de secagem (24 horas). A adsorção da MB seguido do depósito do PPO 300 apresentou maior intervalo linear do que a metodologia do PPY.A comparative study was accomplished between two immobilization methodologies for the meldola's blue (MB), used to decrease the oxidation potential of NADH. The study was performed with glassy carbon electrode by electropolymerization of pyrrole and MB through the technique of cyclic voltammetry being evaluated the effect of the number of cycles to define the thickness and the stability of the film. With 25 cycles it was obtained the best result. They were also investigated the modification of the graphite electrode with film formed by the system sol-gel PPO 300 or PPO 4000 for the dip-coating, through two methods: occlusion and adsorption. The best method was it of adsorption of MB during 10 min, following by deposit of the film PPO 300, favored for the time of drying (24 hours). The adsorption of MB followed by PPO 300 deposition presented broader linear range than PPy methodology

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Silica sol-gel immobilized amperometric biosensor for hydrogen peroxide

Cited by 302 publications

References 21 publications

Kinetic studies of HRP adsorption on ds-DNA immobilized on gold electrode surface by EIS and SPR

Kinetic studies of HRP adsorption on ds-DNA immobilized on gold electrode surface by EIS and SPR

A Biocompatible Nanocomposite for Glucose Sensing

Strategies for developing NADH detector based on meldola blue in different immobilization methods: a comparative study

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