Electrochemical ELISA Protein Biosensing in Undiluted Serum Using a Polypyrrole-Based Platform
An electrochemical enzyme-linked immunosorbent assay (ELISA) biosensor platform using electrochemically prepared ~11 nm thick carboxylic functionalized popypyrrole film has been developed for bio-analyte measurement in undiluted serum. Carboxyl polypyrrole (PPy-COOH) film using 3-carboxy-pyrrol mono...
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doaj-9c23e058a74f42aca0120a119b7873f72020-11-25T03:23:35ZengMDPI AGSensors1424-82202020-05-01202857285710.3390/s20102857Electrochemical ELISA Protein Biosensing in Undiluted Serum Using a Polypyrrole-Based PlatformSunil K. Arya0Pedro Estrela1Department of Electronic & Electrical Engineering, University of Bath, Claverton Down, Bath BA2 7AY, UKDepartment of Electronic & Electrical Engineering, University of Bath, Claverton Down, Bath BA2 7AY, UKAn electrochemical enzyme-linked immunosorbent assay (ELISA) biosensor platform using electrochemically prepared ~11 nm thick carboxylic functionalized popypyrrole film has been developed for bio-analyte measurement in undiluted serum. Carboxyl polypyrrole (PPy-COOH) film using 3-carboxy-pyrrol monomer onto comb-shaped gold electrode microarray (Au) was prepared via cyclic voltammetry (CV). The prepared Au/PPy-COOH was then utilized for electrochemical ELISA platform development by immobilizing analyte-specific antibodies. Tumor necrosis factor-alpha (TNF-α) was selected as a model analyte and detected in undiluted serum. For enhanced performance, the use of a polymeric alkaline phosphatase tag was investigated for the electrochemical ELISA. The developed platform was characterized at each step of fabrication using CV, electrochemical impedance spectroscopy and atomic force microscopy. The bioelectrodes exhibited linearity for TNF-α in the 100 pg/mL–100 ng/mL range when measured in spiked serum, with limit of detection of 78 pg/mL. The sensor showed insignificant signal disturbance from serum proteins and other biologically important proteins. The developed platform was found to be fast and specific and can be applicable for testing and measuring various biologically important protein markers in real samples.https://www.mdpi.com/1424-8220/20/10/2857carboxyl polypyrroleelectrochemical ELISATNF-αpolymeric alkaline phosphatasebiomarkerdifferential pulse voltammetry |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Sunil K. Arya Pedro Estrela |
spellingShingle |
Sunil K. Arya Pedro Estrela Electrochemical ELISA Protein Biosensing in Undiluted Serum Using a Polypyrrole-Based Platform Sensors carboxyl polypyrrole electrochemical ELISA TNF-α polymeric alkaline phosphatase biomarker differential pulse voltammetry |
author_facet |
Sunil K. Arya Pedro Estrela |
author_sort |
Sunil K. Arya |
title |
Electrochemical ELISA Protein Biosensing in Undiluted Serum Using a Polypyrrole-Based Platform |
title_short |
Electrochemical ELISA Protein Biosensing in Undiluted Serum Using a Polypyrrole-Based Platform |
title_full |
Electrochemical ELISA Protein Biosensing in Undiluted Serum Using a Polypyrrole-Based Platform |
title_fullStr |
Electrochemical ELISA Protein Biosensing in Undiluted Serum Using a Polypyrrole-Based Platform |
title_full_unstemmed |
Electrochemical ELISA Protein Biosensing in Undiluted Serum Using a Polypyrrole-Based Platform |
title_sort |
electrochemical elisa protein biosensing in undiluted serum using a polypyrrole-based platform |
publisher |
MDPI AG |
series |
Sensors |
issn |
1424-8220 |
publishDate |
2020-05-01 |
description |
An electrochemical enzyme-linked immunosorbent assay (ELISA) biosensor platform using electrochemically prepared ~11 nm thick carboxylic functionalized popypyrrole film has been developed for bio-analyte measurement in undiluted serum. Carboxyl polypyrrole (PPy-COOH) film using 3-carboxy-pyrrol monomer onto comb-shaped gold electrode microarray (Au) was prepared via cyclic voltammetry (CV). The prepared Au/PPy-COOH was then utilized for electrochemical ELISA platform development by immobilizing analyte-specific antibodies. Tumor necrosis factor-alpha (TNF-α) was selected as a model analyte and detected in undiluted serum. For enhanced performance, the use of a polymeric alkaline phosphatase tag was investigated for the electrochemical ELISA. The developed platform was characterized at each step of fabrication using CV, electrochemical impedance spectroscopy and atomic force microscopy. The bioelectrodes exhibited linearity for TNF-α in the 100 pg/mL–100 ng/mL range when measured in spiked serum, with limit of detection of 78 pg/mL. The sensor showed insignificant signal disturbance from serum proteins and other biologically important proteins. The developed platform was found to be fast and specific and can be applicable for testing and measuring various biologically important protein markers in real samples. |
topic |
carboxyl polypyrrole electrochemical ELISA TNF-α polymeric alkaline phosphatase biomarker differential pulse voltammetry |
url |
https://www.mdpi.com/1424-8220/20/10/2857 |
work_keys_str_mv |
AT sunilkarya electrochemicalelisaproteinbiosensinginundilutedserumusingapolypyrrolebasedplatform AT pedroestrela electrochemicalelisaproteinbiosensinginundilutedserumusingapolypyrrolebasedplatform |
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1724605613645758464 |