A novel immobilization strategy for electrochemical detection of cancer biomarkers: DNA-directed immobilization of aptamer sensors for sensitive detection of prostate specific antigen

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Abstract

We report on a novel strategy for DNA aptamer immobilization to develop sensitive electrochemical detection of protein biomarker, with prostate specific antigen (PSA) as a case biomarker. Thiolated single-stranded DNA was co-immobilized with 3-mercapto-1-propannol on gold electrodes, and used as a scaffold for DNA aptamer attachment through hybridization of the aptamer overhang (so-called “DNA-directed immobilization aptamer sensors”, DDIAS). In the approach, the complementary DNA aptamer against PSA was assembled by the probe ssDNA onto the electrode to detect PSA; or the probe ssDNA directly hybridized with complementary DNA aptamer/PSA complex following their pre-incubation in solution, so-called ‘on-chip’ and ‘in-solution’ method, respectively. A double stranded DNA intercalator with ferrocenyl (Fc) redox marker was synthesized to evaluate the feasibility of the strategy. Results demonstrate that ‘in-solution’ method offers a favourable media (in homogenous solution) for the binding between the aptamer and PSA, which shows to be more efficient than ‘on-chip’ approach. DDIAS show promising analytical performance under optimized conditions, with a limit of detection in the range of fM and low non-specific adsorption.
Original languageEnglish
Pages (from-to)2628-2633
Number of pages6
JournalAnalyst
Volume140
Issue number8
Early online date24 Feb 2015
DOIs
Publication statusPublished - 21 Apr 2015

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Nucleotide Aptamers
Prostate-Specific Antigen
Antigens
Tumor Biomarkers
antigen
Immobilization
immobilization
biomarker
cancer
DNA
sensor
Sensors
Biomarkers
Electrodes
Complementary DNA
Intercalating Agents
Single-Stranded DNA
electrode
Scaffolds
Gold

Cite this

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title = "A novel immobilization strategy for electrochemical detection of cancer biomarkers: DNA-directed immobilization of aptamer sensors for sensitive detection of prostate specific antigen",
abstract = "We report on a novel strategy for DNA aptamer immobilization to develop sensitive electrochemical detection of protein biomarker, with prostate specific antigen (PSA) as a case biomarker. Thiolated single-stranded DNA was co-immobilized with 3-mercapto-1-propannol on gold electrodes, and used as a scaffold for DNA aptamer attachment through hybridization of the aptamer overhang (so-called “DNA-directed immobilization aptamer sensors”, DDIAS). In the approach, the complementary DNA aptamer against PSA was assembled by the probe ssDNA onto the electrode to detect PSA; or the probe ssDNA directly hybridized with complementary DNA aptamer/PSA complex following their pre-incubation in solution, so-called ‘on-chip’ and ‘in-solution’ method, respectively. A double stranded DNA intercalator with ferrocenyl (Fc) redox marker was synthesized to evaluate the feasibility of the strategy. Results demonstrate that ‘in-solution’ method offers a favourable media (in homogenous solution) for the binding between the aptamer and PSA, which shows to be more efficient than ‘on-chip’ approach. DDIAS show promising analytical performance under optimized conditions, with a limit of detection in the range of fM and low non-specific adsorption.",
author = "Zhugen Yang and Barbara Kasprzyk-Hordern and Sean Goggins and Christopher Frost and Pedro Estrela",
year = "2015",
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T1 - A novel immobilization strategy for electrochemical detection of cancer biomarkers: DNA-directed immobilization of aptamer sensors for sensitive detection of prostate specific antigen

AU - Yang, Zhugen

AU - Kasprzyk-Hordern, Barbara

AU - Goggins, Sean

AU - Frost, Christopher

AU - Estrela, Pedro

PY - 2015/4/21

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N2 - We report on a novel strategy for DNA aptamer immobilization to develop sensitive electrochemical detection of protein biomarker, with prostate specific antigen (PSA) as a case biomarker. Thiolated single-stranded DNA was co-immobilized with 3-mercapto-1-propannol on gold electrodes, and used as a scaffold for DNA aptamer attachment through hybridization of the aptamer overhang (so-called “DNA-directed immobilization aptamer sensors”, DDIAS). In the approach, the complementary DNA aptamer against PSA was assembled by the probe ssDNA onto the electrode to detect PSA; or the probe ssDNA directly hybridized with complementary DNA aptamer/PSA complex following their pre-incubation in solution, so-called ‘on-chip’ and ‘in-solution’ method, respectively. A double stranded DNA intercalator with ferrocenyl (Fc) redox marker was synthesized to evaluate the feasibility of the strategy. Results demonstrate that ‘in-solution’ method offers a favourable media (in homogenous solution) for the binding between the aptamer and PSA, which shows to be more efficient than ‘on-chip’ approach. DDIAS show promising analytical performance under optimized conditions, with a limit of detection in the range of fM and low non-specific adsorption.

AB - We report on a novel strategy for DNA aptamer immobilization to develop sensitive electrochemical detection of protein biomarker, with prostate specific antigen (PSA) as a case biomarker. Thiolated single-stranded DNA was co-immobilized with 3-mercapto-1-propannol on gold electrodes, and used as a scaffold for DNA aptamer attachment through hybridization of the aptamer overhang (so-called “DNA-directed immobilization aptamer sensors”, DDIAS). In the approach, the complementary DNA aptamer against PSA was assembled by the probe ssDNA onto the electrode to detect PSA; or the probe ssDNA directly hybridized with complementary DNA aptamer/PSA complex following their pre-incubation in solution, so-called ‘on-chip’ and ‘in-solution’ method, respectively. A double stranded DNA intercalator with ferrocenyl (Fc) redox marker was synthesized to evaluate the feasibility of the strategy. Results demonstrate that ‘in-solution’ method offers a favourable media (in homogenous solution) for the binding between the aptamer and PSA, which shows to be more efficient than ‘on-chip’ approach. DDIAS show promising analytical performance under optimized conditions, with a limit of detection in the range of fM and low non-specific adsorption.

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