Optimisation of an electrochemical impedance spectroscopy aptasensor by exploiting quartz crystal microbalance with dissipation signals
- Formisanoa, Nello, Jolly, Pawan, Bhalla, Nikhil, Cromhout, Mary, Flanagan, Shane P, Fogel, Ronen, Limson, Janice L, Estrela, Pedro
- Authors: Formisanoa, Nello , Jolly, Pawan , Bhalla, Nikhil , Cromhout, Mary , Flanagan, Shane P , Fogel, Ronen , Limson, Janice L , Estrela, Pedro
- Date: 2015
- Subjects: To be catalogued
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/431699 , vital:72797 , xlink:href="https://doi.org/10.1016/j.snb.2015.05.049"
- Description: The response of an Electrochemical Impedance Spectroscopy (EIS) sensor using DNA aptamers is affected by many factors, such as DNA density, charge and conformational changes upon DNA-target binding, and buffer conditions. We report here for the first time on the optimisation of an EIS aptamer-based sensor by using Quartz Crystal Microbalance with Dissipation mode (QCM-D). As a case study, we employed a DNA aptamer against Prostate-Specific Antigen (PSA). PSA detection was achieved by functionalising the gold sensor surface via thiol chemistry with different ratios of thiolated-DNA aptamer and 6-mercapto-1-hexanol (MCH) used as spacer molecules. PSA binding efficiency can be monitored by measuring QCM-D signals which not only provide information about the mass of PSA bound on the sensor surface, but also crucial information about the aptamer conformation and layer hydration.
- Full Text:
- Date Issued: 2015
- Authors: Formisanoa, Nello , Jolly, Pawan , Bhalla, Nikhil , Cromhout, Mary , Flanagan, Shane P , Fogel, Ronen , Limson, Janice L , Estrela, Pedro
- Date: 2015
- Subjects: To be catalogued
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/431699 , vital:72797 , xlink:href="https://doi.org/10.1016/j.snb.2015.05.049"
- Description: The response of an Electrochemical Impedance Spectroscopy (EIS) sensor using DNA aptamers is affected by many factors, such as DNA density, charge and conformational changes upon DNA-target binding, and buffer conditions. We report here for the first time on the optimisation of an EIS aptamer-based sensor by using Quartz Crystal Microbalance with Dissipation mode (QCM-D). As a case study, we employed a DNA aptamer against Prostate-Specific Antigen (PSA). PSA detection was achieved by functionalising the gold sensor surface via thiol chemistry with different ratios of thiolated-DNA aptamer and 6-mercapto-1-hexanol (MCH) used as spacer molecules. PSA binding efficiency can be monitored by measuring QCM-D signals which not only provide information about the mass of PSA bound on the sensor surface, but also crucial information about the aptamer conformation and layer hydration.
- Full Text:
- Date Issued: 2015
Comparison of fluorophore and peroxidase labeled aptamer assays for MUC1 detection in cancer cells
- Flanagan, Shane P, Limson, Janice, Fogel, Ronen
- Authors: Flanagan, Shane P , Limson, Janice , Fogel, Ronen
- Date: 2014
- Subjects: To be catalogued
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/431076 , vital:72742 , xlink:href="10.1109/BioCAS.2014.6981720"
- Description: Aptamers hold great promise for cancer diagnosis and therapy. Several biosensors incorporate aptamers as biorecognition elements for tumor markers although few evaluate their detection in a native conformation and cellular micro-environment. In this study, fluorophore and peroxidase labeled aptamer configurations were compared for the detection of MCF7 breast and SW620 colon cancer cell lines expressing the tumor marker MUC1. Fluorescence based detection showed selective binding to the cell lines relative to a nonbinding control sequence with sequence specific binding differences between MUC1 aptamers accredited to variation in the glycosylation state of expressed MUC1. The peroxidase labeled assay showed high detection sensitivity although low binding specificity was observed for the MUC1 aptamers to the cell lines. Results suggest that aptamers susceptible to non specific binding to cells may limit the applicability of enzymatic amplification to improve aptasensor sensitivity.
- Full Text:
- Date Issued: 2014
- Authors: Flanagan, Shane P , Limson, Janice , Fogel, Ronen
- Date: 2014
- Subjects: To be catalogued
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/431076 , vital:72742 , xlink:href="10.1109/BioCAS.2014.6981720"
- Description: Aptamers hold great promise for cancer diagnosis and therapy. Several biosensors incorporate aptamers as biorecognition elements for tumor markers although few evaluate their detection in a native conformation and cellular micro-environment. In this study, fluorophore and peroxidase labeled aptamer configurations were compared for the detection of MCF7 breast and SW620 colon cancer cell lines expressing the tumor marker MUC1. Fluorescence based detection showed selective binding to the cell lines relative to a nonbinding control sequence with sequence specific binding differences between MUC1 aptamers accredited to variation in the glycosylation state of expressed MUC1. The peroxidase labeled assay showed high detection sensitivity although low binding specificity was observed for the MUC1 aptamers to the cell lines. Results suggest that aptamers susceptible to non specific binding to cells may limit the applicability of enzymatic amplification to improve aptasensor sensitivity.
- Full Text:
- Date Issued: 2014
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