Acetylcholinesterase sensors based on gold electrodes modified with dendrimer and polyaniline: A comparative research

Potentiometric and amperometric enzyme sensors based on modified gold electrodes have been developed and compared in pesticide determination. PAMAM dendrimer (generation G4) stabilized with 1-hexadecanethiol was used for the immobilization of acetylcholinesterase from electric eel and choline oxidase from Alcaligenes species in the assembly of amperometric sensor. Polyaniline-doped with camphorsulfonic acid was used to obtain potentiometric response. Trichlorfon, carbofuran and eserine suppress the biosensor response due to their inhibitory effect. The detection limits of 0.003 and 200 nmol l-1 (trichlorfon), 0.04 and 6 nmol l-1 (carbofuran) and 0.1 and 700 nmol l-1 were obtained for amperometric and potentiometric sensors, respectively. The difference in the biosensor behavior and the high sensitivity of the dendrimer modified sensor to the inhibitors is due to the specific organization of protein layer at charged surface of the modifier macromolecules. © 2004 Elsevier B.V. All rights reserved

Optimization of cytochrome c detection by acoustic and electrochemical methods based on aptamer sensors

© 2016 Elsevier B.V.We report the effect of various factors such as oligonucleotide sequence, buffer composition, ionic strength for optimal determination of cytochrome c (cyt c) by DNA aptamer sensors using thickness shear mode acoustics (TSM) and electrochemical methods. Up to now, several DNA aptamers specific to cyt c have been selected and used in various sensing approaches including optical, electrochemical and mass sensitive transducers. We have analyzed the response of three different aptamers immobilized via biotin-neutravidin method on a gold support by TSM technique. Using this approach we have shown that only 76-length base sequence (apt 76) exhibited specific binding to cyt c with detection limit of 0.50 ± 0.05 nM. This aptamer was then studied under different ionic conditions showing an optimal response for HEPES buffer. Apt 76 based sensor has been also examined by electrochemical methods. However due to the electroactive nature of cyt c, the response of this aptamer was less favorable in comparison with TSM. The apt 76 based sensor was tested also in spiked samples of human plasma by TSM achieving a recovery of 92 ± 6.6% for 1 nM cyt c

Label-Free Toxin Detection by Means of Time-Resolved Electrochemical Impedance Spectroscopy

The real-time detection of trace concentrations of biological toxins requires significant improvement of the detection methods from those reported in the literature. To develop a highly sensitive and selective detection device it is necessary to determine the optimal measuring conditions for the electrochemical sensor in three domains: time, frequency and polarization potential. In this work we utilized a time-resolved electrochemical impedance spectroscopy for the detection of trace concentrations of Staphylococcus enterotoxin B (SEB). An anti-SEB antibody has been attached to the nano-porous aluminum surface using 3-aminopropyltriethoxysilane/glutaraldehyde coupling system. This immobilization method allows fabrication of a highly reproducible and stable sensing device. Using developed immobilization procedure and optimized detection regime, it is possible to determine the presence of SEB at the levels as low as 10 pg/mL in 15 minutes

Near-Infrared Fluorescence Detection of Acetylcholine in Aqueous Solution Using a Complex of Rhodamine 800 and p-Sulfonato-calix[8]arene

The complexing properties of p-sulfonatocalix[n]arenes (n = 4: S[4], n = 6: S[6], and n = 8: S[8]) for rhodamine 800 (Rh800) and indocyanine green (ICG) were examined to develop a near-infrared (NIR) fluorescence detection method for acetylcholine (ACh). We found that Rh800 (as a cation) forms an inclusion complex with S[n], while ICG (as a twitter ion) have no binding ability for S[n]. The binding ability of Rh800 to S[n] decreased in the order of S[8] > S[6] >> S[4]. By the formation of the complex between Rh800 and S[8], fluorescence intensity of the Rh800 was significantly decreased. From the fluorescence titration of Rh800 by S[8], stoichiometry of the Rh800-S[8] complex was determined to be 1:1 with a dissociation constant of 2.2 μM in PBS. The addition of ACh to the aqueous solution of the Rh800-S[8] complex caused a fluorescence increase of Rh800, resulting from a competitive replacement of Rh800 by ACh in the complex. From the fluorescence change by the competitive fluorophore replacement, stoichiometry of the Rh800-ACh complex was found to be 1:1 with a dissociation constant of 1.7 mM. The effects of other neurotransmitters on the fluorescence spectra of the Rh800-S[8] complex were examined for dopamine, GABA, glycine, and l-asparatic acid. Among the neurotransmitters examined, fluorescence response of the Rh800-S[8] complex was highly specific to ACh. Rh800-S[8] complexes can be used as a NIR fluorescent probe for the detection of ACh (5 × 10−4−10−3 M) in PBS buffer (pH = 7.2)

DNA Hybridization Sensors Based on Electrochemical Impedance Spectroscopy as a Detection Tool

Recent advances in label free DNA hybridization sensors employing electrochemical impedance spectroscopy ( EIS) as a detection tool are reviewed. These sensors are based on the modulation of the blocking ability of an electrode modified with a probe DNA by an analyte, i.e., target DNA. The probe DNA is immobilized on a self-assembled monolayer, a conducting polymer film, or a layer of nanostructures on the electrode such that desired probe DNA would selectively hybridize with target DNA. The rate of charge transfer from the electrode thus modified to a redox indicator, e. g., [Fe(CN)(6)](3-/4-), which is measured by EIS in the form of charge transfer resistance (R(ct)), is modulated by whether or not, as well as how much, the intended target DNA is selectively hybridized. Efforts made to enhance the selectivity as well as the sensitivity of DNA sensors and to reduce the EIS measurement time are briefly described along with brief future perspectives in developing DNA sensors.open484

Acetylcholine biosensor based on dendrimer layers for pesticides detection

We tested a new design of an enzyme biosensor based on acetylcholinesterase (AChE) and choline oxidase (ChO) immobilized on the supported monomolecular layer composed of poly(amidoamine) (PAMAM) dendrimers of the fourth generation (G4) mixed with 1-hexadecanethiol (HDT). The resulting enzymatic activity, measured amperometrically, was substantially depressed in the presence of the organophosphate pesticide dimethyl-2,2-dichlorovinylphosphate (DDVP, Dichlorvos), carbamate pesticides carbofuran and carbamate drug eserine. The detection limits (1.3 × 10 -3 ppb for DDVP, 0.01 ppb for carbofuran and 0.03 for eserine) were considerably lower than so far reported for AChE based amperometric and potentiometric sensors. The relative simple protocol of biosensor preparation, high sensitivity and stability is very promising for determination of environmental pollutants in field conditions

Optimization of cytochrome c detection by acoustic and electrochemical methods based on aptamer sensors

© 2016 Elsevier B.V.We report the effect of various factors such as oligonucleotide sequence, buffer composition, ionic strength for optimal determination of cytochrome c (cyt c) by DNA aptamer sensors using thickness shear mode acoustics (TSM) and electrochemical methods. Up to now, several DNA aptamers specific to cyt c have been selected and used in various sensing approaches including optical, electrochemical and mass sensitive transducers. We have analyzed the response of three different aptamers immobilized via biotin-neutravidin method on a gold support by TSM technique. Using this approach we have shown that only 76-length base sequence (apt 76) exhibited specific binding to cyt c with detection limit of 0.50 ± 0.05 nM. This aptamer was then studied under different ionic conditions showing an optimal response for HEPES buffer. Apt 76 based sensor has been also examined by electrochemical methods. However due to the electroactive nature of cyt c, the response of this aptamer was less favorable in comparison with TSM. The apt 76 based sensor was tested also in spiked samples of human plasma by TSM achieving a recovery of 92 ± 6.6% for 1 nM cyt c

Optimization of cytochrome c detection by acoustic and electrochemical methods based on aptamer sensors

© 2016 Elsevier B.V.We report the effect of various factors such as oligonucleotide sequence, buffer composition, ionic strength for optimal determination of cytochrome c (cyt c) by DNA aptamer sensors using thickness shear mode acoustics (TSM) and electrochemical methods. Up to now, several DNA aptamers specific to cyt c have been selected and used in various sensing approaches including optical, electrochemical and mass sensitive transducers. We have analyzed the response of three different aptamers immobilized via biotin-neutravidin method on a gold support by TSM technique. Using this approach we have shown that only 76-length base sequence (apt 76) exhibited specific binding to cyt c with detection limit of 0.50 ± 0.05 nM. This aptamer was then studied under different ionic conditions showing an optimal response for HEPES buffer. Apt 76 based sensor has been also examined by electrochemical methods. However due to the electroactive nature of cyt c, the response of this aptamer was less favorable in comparison with TSM. The apt 76 based sensor was tested also in spiked samples of human plasma by TSM achieving a recovery of 92 ± 6.6% for 1 nM cyt c

Optimization of cytochrome c detection by acoustic and electrochemical methods based on aptamer sensors

© 2016 Elsevier B.V.We report the effect of various factors such as oligonucleotide sequence, buffer composition, ionic strength for optimal determination of cytochrome c (cyt c) by DNA aptamer sensors using thickness shear mode acoustics (TSM) and electrochemical methods. Up to now, several DNA aptamers specific to cyt c have been selected and used in various sensing approaches including optical, electrochemical and mass sensitive transducers. We have analyzed the response of three different aptamers immobilized via biotin-neutravidin method on a gold support by TSM technique. Using this approach we have shown that only 76-length base sequence (apt 76) exhibited specific binding to cyt c with detection limit of 0.50 ± 0.05 nM. This aptamer was then studied under different ionic conditions showing an optimal response for HEPES buffer. Apt 76 based sensor has been also examined by electrochemical methods. However due to the electroactive nature of cyt c, the response of this aptamer was less favorable in comparison with TSM. The apt 76 based sensor was tested also in spiked samples of human plasma by TSM achieving a recovery of 92 ± 6.6% for 1 nM cyt c