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

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

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

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

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

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