Study of zinc hexacyanoferrate-modified platinum electrodes using electrochemical quartz crystal microbalance

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Quartz Crystal Microbalance monitoring the real-time binding of lectin with carbohydrate with high and low molecular mass

Quartz Crystal Microbalance (QCM) was used to monitor the mass changes on a quartz crystal surface containing immobilized lectins that interacted with carbohydrates. The strategy for lectin immobilization was developed on the basis of a multilayer system composed of Au-cystamine-glutaraldehyde-lectin. Each step of the immobilization procedure was confirmed by FTIR analysis. The system was used to study the interactions of Concanavalin A (ConA) with maltose and Jacalin with Fetuin. The real-time binding of different concentrations of carbohydrate to the immobilized lectin was monitored by means of QCM measurements and the data obtained allowed for the construction of Langmuir isotherm curves. The association constants determined for the specific interactions analyzed here were (6.4 +/- 0.2) X 10(4) M-1 for Jacalin-Fetuin and (4.5 +/- 0.1) x 10(2) M-1 for ConA-maltose. These results indicate that the QCM constitutes a suitable method for the analysis of lectin-carbohydrate interactions, even when assaying low molecular mass ligands such as disaccharides. Published by Elsevier B.V.

Quartz crystal microbalance as a tool for kinetic enzymatic assays by variation of pH

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Electrogravimetric Analysis by Quartz-Crystal Microbalance on the Consumption of the Neurotransmitter Acetylcholine by Acetylcholinesterase

Electrogravimetric analysis was performed on the consumption of the neurotransmitter Acetylcholine (ACh) by Acetylcholinesterase (AChE) in situ and in real time. Michaelis-Menten assumption was achieved by using an enzyme micro-reactor in which the total enzyme was anchored in a quartz crystal microbalance chip (QCM-chip) with a strategically engineered self-assembled monolayer (SAM) of alkanethiols, which can prevent diffusion-controlled or spatially restricted kinetics. The real-time frequency changes indicated the rate of the products formation from enzymatic reaction. The QCM-chip was tested showing that it could demonstrate AChE inhibition by physostigmine. © 2013 Copyright Taylor and Francis Group, LLC.

Mass/charge balance as a tool to estimate dimensional change in polypyrrole-based actuators

The deconvolution of the voltammograms of polypyrrole electrochemistry has proved to be possible through the electrochemical quartz crystal microbalance data using the F(dm/dQ) function. This deconvolution allows the evolution of the thickness of the polypyrrole films during their redox processes to be estimated and therefore, the mechanical contraction/decontraction of this polymer as a function of the ionic exchange processes can be evaluated. (c) 2005 Elsevier B.V. All rights reserved.

EQCM study during lithium insertion/deinsertion processes in Nb2O5 films prepared by polymeric precursor method

The electrochemical quartz crystal microbalance (EQCM) technique was used to study two chemically distinct Nb2O5 electrochromic thin films (one pure and the other lithium-doped) during the lithium electroinsertion reaction. In the initial cycles, the electrode showed an irreversible mass variation greater than expected for Li+ insertion/deinsertion processes, which was attributed to the wettability effect (allied to the porous morphology) that emerged as the dominant process in apparent electrode mass changes. As the cycles progressed, the mass variation stabilized and the changes in apparent mass became reversible, showing a good correlation with the charge variations.The results generally indicated that the Li+ insertion/deinsertion process occurred more easily in the Nb2O5-doped film, which also displayed a greater capacity for Li+ insertion. However, a total mass/charge balance analysis revealed that the stoichiometry of the Li+ solid state insertion/deinsertion reaction was similar in the two electrodes under study. © 2005 Elsevier B.V. All rights reserved.

Detecção de anilinas substituídas, em filmes de hexacianoferratos, com microbalança de cristal de quartzo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo de processos redox enzimáticos confinados em eletrodos pelo concomitante monitoramento da variação de massa e da corrente: o caso da penicilinase como modelo

Pós-graduação em Biotecnologia - IQ

Determinação da constante de afinidade e cinética da interação lectina ArtinM-célula leucêmica (NB4) por meio de técnicas piezoelétrica e eletroquímica

Pós-graduação em Biotecnologia - IQ

Evaluating the equilibrium association constant between artinM lectin and myeloid leukemia cells by impedimetric and piezoelectric label free approaches

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Physical Vapor Deposited Thin Films of Lignins Extracted from Sugar Cane Bagasse: Morphology, Electrical Properties, and Sensing Applications

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Spray layer-by-layer films based on phospholipid vesicles aiming sensing application via e-tongue system

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Hydration effects on the structural properties and haem-haem interaction in haemoglobin

Direct and simultaneous measurements of hydration water content and protein conformation have been performed using quartz crystal microbalance and visible absorption spectroscopy. Equilibrium and kinetics of methaemoglobin/haemichrome transition induced by the alteration of the degree of hydration was investigated in thin films exposed to controlled humidity. The kinetics experiment show that the conversion of species achieve the equilibrium more rapidly that the amount of sorbed water by the protein. The transition shows a sigmoid behaviour and suggest cooperative phenomena manifested by haem-haem interaction. The water hydration network contributing to the haem haem interaction advise that water acts as allosteric effectors for the conversion between species. Irreversible changes produced by complete drying are clearly shown.

The role of hydration on the mechanism of allosteric regulation: In situ measurements of the oxygen-linked kinetics of water binding to hemoglobin

We report here the first direct measurements of changes in protein hydration triggered by a functional binding. This task is achieved by weighing hemoglobin (Hb) and myoglobin films exposed to an atmosphere of 98%, relative humidity during oxygenation. The binding of the first oxygen molecules to Hb tetramer triggers a change in protein conformation, which increases binding affinity to the remaining empty sites giving rise to the appearance of cooperative phenomena. Although crystallographic data have evidenced that this structural change increases the protein water-accessible surface area, isobaric osmotic stress experiments in aqueous cosolutions have shown that water binding is linked to Hb oxygenation. Now we show that the differential hydration between fully oxygenated and fully deoxygenated states of these proteins, determined by weighing protein films with a quartz crystal microbalance, agree with the ones determined by osmotic stress in aqueous cosolutions, from the linkage between protein oxygen affinity and water activity. The agreements prove that the changes in water activity brought about by adding osmolytes to the buffer solution shift biochemical equilibrium in proportion to the number of water molecules associated with the reaction. The concomitant kinetics of oxygen and of water binding to Hb have been also determined. The data show that the binding of water molecules to the extra protein surface exposed on the transition from the low-affinity T to the high-affinity R conformations of hemoglobin is the rate-limiting step of Hb cooperative reaction. This evidences that water binding is a crucial step on the allosteric mechanism regulating cooperative interactions, and suggests the possibility that environmental water activity might be engaged in the kinetic control of some important reactions in vivo.

Real-time monitoring and kinetic parameter estimation of the affinity interaction of jArtinM and rArtinM with peroxidase glycoprotein by the electrogravimetric technique

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)