8-hydroxy-2′-deoxyguanosine (8-OHdG) biomarker detection down to picoMolar level on a plastic antibody film

Gabriela V. Martins, Ana Cláudia Lourenço Santana Marques, Elvira Fortunato, M. Goreti F Sales

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

An innovative biosensor assembly relying on a simple and straightforward in-situ construction is presented to monitor urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) down to the pmol/L level. The sensing film of the biosensor consisted of a molecularly imprinted polymer (MIP) layer for 8-OHdG assembled on a gold electrode through electropolymerization of monomer combined with the template. The analytical features of the resulting biosensor were assessed by Cyclic Voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS). Some experimental parameters such as the initial concentration of the monomer and the ratio template-monomer were investigated and optimized in order to finely tune the performance of the MIP-based sensor. Under optimal conditions, the developed biosensor was able to rebind 8-OHdG with a linear response against EIS from 0.1 to 100 pg/ml 3.5-3500 pM. The interference of coexisting species was tested, also with calibrations on urine samples, and good selectivity towards 8-OHdG was obtained. RAMAN spectroscopy, FTIR and SEM evaluations of the prepared films confirmed the formation of a polyphenol thin-film on the electrode surface. The presence and distribution of the imprinted cavities on the MIP layer was confirmed by confocal microscopy imaging of the film, after a post-treatment with Fluorescein Isothiocyanate (FITC) labeled 8-OHdG antibody. Overall, this label-free biosensor for urinary 8-OHdG detection constitutes a promising low-cost alternative to the conventional immunoassay approaches, due to its simplicity, stability, high sensitivity and selectivity for biological sample assays, opening new doors for other applications.

Original languageEnglish
Pages (from-to)225-234
Number of pages10
JournalBiosensors & Bioelectronics
Volume86
DOIs
Publication statusPublished - 15 Dec 2016

Fingerprint

Biosensing Techniques
Biomarkers
Biosensors
Antibodies
Plastics
Dielectric Spectroscopy
Polymers
Monomers
Electrochemical impedance spectroscopy
Electrodes
Electropolymerization
Confocal microscopy
Polyphenols
Fourier Transform Infrared Spectroscopy
Fluorescein
Immunoassay
Confocal Microscopy
Gold
Biological Assay
Calibration

Keywords

  • MOLECULARLY IMPRINTED POLYMERS
  • OXIDATIVE DNA-DAMAGE
  • SCREEN-PRINTED ELECTRODES
  • GLASSY-CARBON ELECTRODE
  • ELECTROCHEMICAL SENSOR
  • URINARY 8-HYDROXY-2'-DEOXYGUANOSINE
  • CAPILLARY-ELECTROPHORESIS
  • GOLD NANOPARTICLES
  • CANCER-PATIENTS
  • STRESS

Cite this

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title = "8-hydroxy-2′-deoxyguanosine (8-OHdG) biomarker detection down to picoMolar level on a plastic antibody film",
abstract = "An innovative biosensor assembly relying on a simple and straightforward in-situ construction is presented to monitor urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) down to the pmol/L level. The sensing film of the biosensor consisted of a molecularly imprinted polymer (MIP) layer for 8-OHdG assembled on a gold electrode through electropolymerization of monomer combined with the template. The analytical features of the resulting biosensor were assessed by Cyclic Voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS). Some experimental parameters such as the initial concentration of the monomer and the ratio template-monomer were investigated and optimized in order to finely tune the performance of the MIP-based sensor. Under optimal conditions, the developed biosensor was able to rebind 8-OHdG with a linear response against EIS from 0.1 to 100 pg/ml 3.5-3500 pM. The interference of coexisting species was tested, also with calibrations on urine samples, and good selectivity towards 8-OHdG was obtained. RAMAN spectroscopy, FTIR and SEM evaluations of the prepared films confirmed the formation of a polyphenol thin-film on the electrode surface. The presence and distribution of the imprinted cavities on the MIP layer was confirmed by confocal microscopy imaging of the film, after a post-treatment with Fluorescein Isothiocyanate (FITC) labeled 8-OHdG antibody. Overall, this label-free biosensor for urinary 8-OHdG detection constitutes a promising low-cost alternative to the conventional immunoassay approaches, due to its simplicity, stability, high sensitivity and selectivity for biological sample assays, opening new doors for other applications.",
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8-hydroxy-2′-deoxyguanosine (8-OHdG) biomarker detection down to picoMolar level on a plastic antibody film. / Martins, Gabriela V.; Marques, Ana Cláudia Lourenço Santana; Fortunato, Elvira; Sales, M. Goreti F.

In: Biosensors & Bioelectronics, Vol. 86, 15.12.2016, p. 225-234.

Research output: Contribution to journalArticle

TY - JOUR

T1 - 8-hydroxy-2′-deoxyguanosine (8-OHdG) biomarker detection down to picoMolar level on a plastic antibody film

AU - Martins, Gabriela V.

AU - Marques, Ana Cláudia Lourenço Santana

AU - Fortunato, Elvira

AU - Sales, M. Goreti F

N1 - Sem PDF conforme despacho. info:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBD%2F94159%2F2013/PT# info:eu-repo/grantAgreement/FCT/5876/147333/PT# Portuguese Foundation for Science and Technology (FCT) (SFRH/BD/94159/2013) FEDER funds through the COMPETE Programme National Funds (UID/CTM/50025/2013)

PY - 2016/12/15

Y1 - 2016/12/15

N2 - An innovative biosensor assembly relying on a simple and straightforward in-situ construction is presented to monitor urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) down to the pmol/L level. The sensing film of the biosensor consisted of a molecularly imprinted polymer (MIP) layer for 8-OHdG assembled on a gold electrode through electropolymerization of monomer combined with the template. The analytical features of the resulting biosensor were assessed by Cyclic Voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS). Some experimental parameters such as the initial concentration of the monomer and the ratio template-monomer were investigated and optimized in order to finely tune the performance of the MIP-based sensor. Under optimal conditions, the developed biosensor was able to rebind 8-OHdG with a linear response against EIS from 0.1 to 100 pg/ml 3.5-3500 pM. The interference of coexisting species was tested, also with calibrations on urine samples, and good selectivity towards 8-OHdG was obtained. RAMAN spectroscopy, FTIR and SEM evaluations of the prepared films confirmed the formation of a polyphenol thin-film on the electrode surface. The presence and distribution of the imprinted cavities on the MIP layer was confirmed by confocal microscopy imaging of the film, after a post-treatment with Fluorescein Isothiocyanate (FITC) labeled 8-OHdG antibody. Overall, this label-free biosensor for urinary 8-OHdG detection constitutes a promising low-cost alternative to the conventional immunoassay approaches, due to its simplicity, stability, high sensitivity and selectivity for biological sample assays, opening new doors for other applications.

AB - An innovative biosensor assembly relying on a simple and straightforward in-situ construction is presented to monitor urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) down to the pmol/L level. The sensing film of the biosensor consisted of a molecularly imprinted polymer (MIP) layer for 8-OHdG assembled on a gold electrode through electropolymerization of monomer combined with the template. The analytical features of the resulting biosensor were assessed by Cyclic Voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS). Some experimental parameters such as the initial concentration of the monomer and the ratio template-monomer were investigated and optimized in order to finely tune the performance of the MIP-based sensor. Under optimal conditions, the developed biosensor was able to rebind 8-OHdG with a linear response against EIS from 0.1 to 100 pg/ml 3.5-3500 pM. The interference of coexisting species was tested, also with calibrations on urine samples, and good selectivity towards 8-OHdG was obtained. RAMAN spectroscopy, FTIR and SEM evaluations of the prepared films confirmed the formation of a polyphenol thin-film on the electrode surface. The presence and distribution of the imprinted cavities on the MIP layer was confirmed by confocal microscopy imaging of the film, after a post-treatment with Fluorescein Isothiocyanate (FITC) labeled 8-OHdG antibody. Overall, this label-free biosensor for urinary 8-OHdG detection constitutes a promising low-cost alternative to the conventional immunoassay approaches, due to its simplicity, stability, high sensitivity and selectivity for biological sample assays, opening new doors for other applications.

KW - MOLECULARLY IMPRINTED POLYMERS

KW - OXIDATIVE DNA-DAMAGE

KW - SCREEN-PRINTED ELECTRODES

KW - GLASSY-CARBON ELECTRODE

KW - ELECTROCHEMICAL SENSOR

KW - URINARY 8-HYDROXY-2'-DEOXYGUANOSINE

KW - CAPILLARY-ELECTROPHORESIS

KW - GOLD NANOPARTICLES

KW - CANCER-PATIENTS

KW - STRESS

UR - http://www.scopus.com/inward/record.url?scp=84976863902&partnerID=8YFLogxK

U2 - 10.1016/j.bios.2016.06.052

DO - 10.1016/j.bios.2016.06.052

M3 - Article

VL - 86

SP - 225

EP - 234

JO - Biosensors & Bioelectronics

JF - Biosensors & Bioelectronics

SN - 0956-5663

ER -