Investigation of the interaction between Tc85-11 protein and antibody anti-T. cruzi by AFM and amperometric measurements

Ferreira, Antonio Aparecido Pupim; Colli, Walter; Alves, Maria Júlia M.; Oliveira, Daniel R.; Costa, Paulo Inácio da; Güell, Aleix G.; Sanz, Fausto; Benedetti, Assis Vicente; Yamanaka, Hideko

doi:10.1016/j.electacta.2006.03.061

Cited by 20 publications

(29 citation statements)

References 20 publications

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“…The roughness of the surfaces, R q , obtained by averaging over 20 different regions with area of 200 Â 200 nm 2 is 0.52 AE 0.08 nm. These features are in a very good agreement with those reported in literature for the same kind of samples (Ferreira et al, 2006). Upon incubating this functionalized gold substrate with a solution containing DBD for 2 h and rinsing with milliQ water, bright large spots randomly distributed over the surface appear in the TM-AFM images recorded in milliQ water (see Figure 2B).…”

Section: Afm Imaging Experimentssupporting

confidence: 87%

A combined atomic force microscopy imaging and docking study to investigate the complex between p53 DNA binding domain and Azurin

Bizzarri

Agostino²,

Andolfi

et al. 2009

J of Molecular Recognition

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The tumor suppressor p53 interacts with the redox copper protein Azurin (AZ) forming a complex which is of some relevance in biomedicine and cancer therapy. To obtain information on the spatial organization of this complex when it is immobilized on a substrate, we have used tapping mode-atomic force microscopy (TM-AFM) imaging combined with computational docking. The vertical dimension and the bearing volume of the DNA binding domain (DBD) of p53, anchored to functionalized gold substrate through exposed lysine residues, alone and after deposing AZ, have been measured by TM-AFM. By a computational docking approach, a three-dimensional model for the DBD of p53, before and after addition of AZ, have been predicted. Then we have calculated the possible arrangements of these biomolecular systems on gold substrate by finding a good agreement with the related experimental distribution of the height. The potentiality of the approach combining TM-AFM imaging and computational docking for the study of biomolecular complexes immobilized on substrates is briefly discussed.

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Section: Afm Imaging Experimentssupporting

confidence: 87%

A combined atomic force microscopy imaging and docking study to investigate the complex between p53 DNA binding domain and Azurin

Bizzarri

Agostino²,

Andolfi

et al. 2009

J of Molecular Recognition

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“…Consequently, the reduction of species A on the electrode surface can be used to follow the peroxidase catalyzed reaction [8,30,33,[35][36][37]. In our work, the HRP is monitored by using hydrogen peroxide as enzyme substrate and iodide as mediator (AH).…”

Section: Amperometric Monitoring Of Enzymatic Reactionmentioning

confidence: 99%

“…After antigen immobilization on the CDtrode surface, the immunoassay was performed using the following procedure ( Figure 1): a) blocking of the sensor surface with 50 mL of PBS-T buffer solution containing 0.5 % bovine serum albumin and incubation for 40 min at room temperature; b) washing in Milli-Q water; c) application of 50 mL of T. cruzi positive serum (Ab) diluted in blocking solution and incubation for 15 min; d) washing in Milli-Q water; e) application of 50 mL of the anti-human IgG antibody conjugated to peroxidase (Ab*) diluted in blocking solution and incubation for 15 min; f) washing in Milli-Q water; g) monitoring immunoenzymatic reaction by amperometric technique in hydrogen peroxide (substrate) and potassium iodide (mediator) solutions in the proportion of 1 : 3 [30] respectively, prepared in 0.1 mol L À1 phosphate buffer solution at pH 7.0 and incubation for 10 min.…”

Section: Preparation Of Immunosensormentioning

confidence: 99%

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Amperometric Immunosensor for Chagas’ Disease Using Gold CD‐R Transducer

et al. 2011

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Chagas disease has been spread mainly through blood transfusion or blood components, therefore the development of a device for neglected diseases with high specificity and sensitivity is important to public health. This work describes the development of an amperometric immunosensor for determination of Chagas disease through a goldbased electrode obtained from a recordable compact disc modified with 4-(methylmercapto)benzaldehyde for the immobilization of Tc85 protein of the Trypanosoma cruzi. The immunoassays were carried out using positive and negative sera from Chagasdisease patients and immunoglobulin conjugated with peroxidase enzyme. The immunosensor presented À0.949 mA as cut-off value and was applied in sera samples.

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“…A molécula do mediador escolhido deve reagir rapidamente com a peroxidase oxidada. Uma variedade de moléculas doadoras de elétrons pode reagir rapidamente com os compostos I e II, sendo que, para a enzima HRP pode-se utilizar como mediador de elétrons: hidroquinona [9,10], ácido ascórbico [11], ferroceno [12,13], iodeto [14,15,16], entre outros. O ácido 5-aminossalicílico (5-ASA) também tem sido usado como mediador para a enzima HRP em imunoensaios com detecção ótica [17], potenciométrica [18] e amperométrica [19,20,21], sendo que nos trabalhos utilizando a técnica amperométrica, a HRP foi imobilizada na superfície dos diferentes eletrodos.…”

Section: (5)unclassified

Estudo do comportamento eletroquímico da enzima peroxidase na presença de peróxido de hidrogênio e ácido 5-aminossalicílico

2008

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The electrochemical behavior of the enzyme peroxidase (HRP) was investigated using the hydrogen peroxide as enzymatic substrate and the 5-aminosalicylic acid (5-ASA) as mediator of electrons on graphite electrodes. Several parameters were optimized, namely, the applied potential to the amperometric technique fixed in -0.125V, the 0.1 mol L-1 phosphate-citrate buffer at pH 5.0 as supporting electrolyte and the proportion between the 5-ASA and H2O2 in 1:7, among others. It was observed the catalysis of the oxidation reaction of the H2O2 in the presence of the enzyme HRP and 5-ASA. The oxidation product was reduced in the electrode surface, evidencing a significant increase in the intensity of the cathodic current.

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Investigation of the interaction between Tc85-11 protein and antibody anti-T. cruzi by AFM and amperometric measurements

Cited by 20 publications

References 20 publications

A combined atomic force microscopy imaging and docking study to investigate the complex between p53 DNA binding domain and Azurin

A combined atomic force microscopy imaging and docking study to investigate the complex between p53 DNA binding domain and Azurin

Amperometric Immunosensor for Chagas’ Disease Using Gold CD‐R Transducer

Estudo do comportamento eletroquímico da enzima peroxidase na presença de peróxido de hidrogênio e ácido 5-aminossalicílico

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