Covalent Hemin/G4 complex-linked sandwich bioassay on magnetic beads for femtomolar HER-2/neu detection in human serum via direct electrocatalytic reduction of oxygen

Ana Díaz-Fernández; Elena E. Ferapontova

doi:10.1016/j.aca.2022.340049

Covalent Hemin/G4 complex-linked sandwich bioassay on magnetic beads for femtomolar HER-2/neu detection in human serum via direct electrocatalytic reduction of oxygen

Ana Díaz-Fernández, Elena E. Ferapontova^*

^*Corresponding author for this work

Interdisciplinary Nanoscience Center

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

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Abstract

Liquid biopsy assays for tumour biomarkers circulating in blood are perspective non-invasive tools for cancer diagnosis and treatment monitoring. Here, we suggest a simple, 1 h long electrochemical DNAzyme-linked aptamer- and immuno-sandwich magnetic assay for analysis of serum HER-2/neu protein overexpressed in several aggressive cancers. In the assay, we used a covalent hemin-guanine quadruplex (G4) complex as a novel O₂-dependent electrocatalytic label that allowed 10 fM (aptamer-aptamer) and 1 fM (aptamer-antibody) detection of HER-2/neu in human serum. The O₂ reactivity of the aptamer-conjugated label was detected at high-surface-area graphite electrodes displaying a high efficiency of O₂ reduction electro-catalyzed by this DNAzyme. In contrast to the recognised H₂O₂ reactivity, the O₂ reactivity of the covalent hemin/G4 complex depended only on ambient O₂ present in solutions, and did not require adding such traditional reagents as hemin and H₂O₂, and solution de-aeration. Human serum albumin, urokinase plasminogen activator and thrombin did not interfere, and the assay was used for analysis of basal serum levels of HER-2/neu. Due to the simplicity and low cost, sandwich assays exploiting O₂-linked electrocatalysis by the covalent hemin-G4 complexes represent a more advanced electrochemical ELISA platform for ultrasensitive and fast detection of low concentrations of proteins in complex biological matrices.

Original language	English
Article number	340049
Journal	Analytica Chimica Acta
Volume	1219
Number of pages	7
ISSN	0003-2670
DOIs	https://doi.org/10.1016/j.aca.2022.340049
Publication status	Published - Aug 2022

Keywords

Antibody
Aptamer
Covalent Hemin/G4 complex
DNAzyme
Electrochemical sandwich assay
HER-2/neu
Magnetic beads
O electrocatalysis
Limit of Detection
Oxygen
Hydrogen Peroxide/chemistry
Humans
Hemin/chemistry
DNA, Catalytic/chemistry
Magnetic Phenomena
Biosensing Techniques
Biological Assay
G-Quadruplexes
Aptamers, Nucleotide/chemistry

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10.1016/j.aca.2022.340049Licence: CC BY

1-s2.0-S0003267022006201-mainFinal published version, 3.61 MBLicence: CC BY

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@article{9493f93ad070439688395f5cb648b4a4,

title = "Covalent Hemin/G4 complex-linked sandwich bioassay on magnetic beads for femtomolar HER-2/neu detection in human serum via direct electrocatalytic reduction of oxygen",

abstract = "Liquid biopsy assays for tumour biomarkers circulating in blood are perspective non-invasive tools for cancer diagnosis and treatment monitoring. Here, we suggest a simple, 1 h long electrochemical DNAzyme-linked aptamer- and immuno-sandwich magnetic assay for analysis of serum HER-2/neu protein overexpressed in several aggressive cancers. In the assay, we used a covalent hemin-guanine quadruplex (G4) complex as a novel O2-dependent electrocatalytic label that allowed 10 fM (aptamer-aptamer) and 1 fM (aptamer-antibody) detection of HER-2/neu in human serum. The O2 reactivity of the aptamer-conjugated label was detected at high-surface-area graphite electrodes displaying a high efficiency of O2 reduction electro-catalyzed by this DNAzyme. In contrast to the recognised H2O2 reactivity, the O2 reactivity of the covalent hemin/G4 complex depended only on ambient O2 present in solutions, and did not require adding such traditional reagents as hemin and H2O2, and solution de-aeration. Human serum albumin, urokinase plasminogen activator and thrombin did not interfere, and the assay was used for analysis of basal serum levels of HER-2/neu. Due to the simplicity and low cost, sandwich assays exploiting O2-linked electrocatalysis by the covalent hemin-G4 complexes represent a more advanced electrochemical ELISA platform for ultrasensitive and fast detection of low concentrations of proteins in complex biological matrices.",

keywords = "Antibody, Aptamer, Covalent Hemin/G4 complex, DNAzyme, Electrochemical sandwich assay, HER-2/neu, Magnetic beads, O electrocatalysis, Limit of Detection, Oxygen, Hydrogen Peroxide/chemistry, Humans, Hemin/chemistry, DNA, Catalytic/chemistry, Magnetic Phenomena, Biosensing Techniques, Biological Assay, G-Quadruplexes, Aptamers, Nucleotide/chemistry",

author = "Ana D{\'i}az-Fern{\'a}ndez and Ferapontova, {Elena E.}",

year = "2022",

month = aug,

doi = "10.1016/j.aca.2022.340049",

language = "English",

volume = "1219",

journal = "Analytica Chimica Acta",

issn = "0003-2670",

publisher = "Elsevier B.V.",

}

Covalent Hemin/G4 complex-linked sandwich bioassay on magnetic beads for femtomolar HER-2/neu detection in human serum via direct electrocatalytic reduction of oxygen. / Díaz-Fernández, Ana; Ferapontova, Elena E.
In: Analytica Chimica Acta, Vol. 1219, 340049, 08.2022.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - Covalent Hemin/G4 complex-linked sandwich bioassay on magnetic beads for femtomolar HER-2/neu detection in human serum via direct electrocatalytic reduction of oxygen

AU - Díaz-Fernández, Ana

AU - Ferapontova, Elena E.

PY - 2022/8

Y1 - 2022/8

N2 - Liquid biopsy assays for tumour biomarkers circulating in blood are perspective non-invasive tools for cancer diagnosis and treatment monitoring. Here, we suggest a simple, 1 h long electrochemical DNAzyme-linked aptamer- and immuno-sandwich magnetic assay for analysis of serum HER-2/neu protein overexpressed in several aggressive cancers. In the assay, we used a covalent hemin-guanine quadruplex (G4) complex as a novel O2-dependent electrocatalytic label that allowed 10 fM (aptamer-aptamer) and 1 fM (aptamer-antibody) detection of HER-2/neu in human serum. The O2 reactivity of the aptamer-conjugated label was detected at high-surface-area graphite electrodes displaying a high efficiency of O2 reduction electro-catalyzed by this DNAzyme. In contrast to the recognised H2O2 reactivity, the O2 reactivity of the covalent hemin/G4 complex depended only on ambient O2 present in solutions, and did not require adding such traditional reagents as hemin and H2O2, and solution de-aeration. Human serum albumin, urokinase plasminogen activator and thrombin did not interfere, and the assay was used for analysis of basal serum levels of HER-2/neu. Due to the simplicity and low cost, sandwich assays exploiting O2-linked electrocatalysis by the covalent hemin-G4 complexes represent a more advanced electrochemical ELISA platform for ultrasensitive and fast detection of low concentrations of proteins in complex biological matrices.

AB - Liquid biopsy assays for tumour biomarkers circulating in blood are perspective non-invasive tools for cancer diagnosis and treatment monitoring. Here, we suggest a simple, 1 h long electrochemical DNAzyme-linked aptamer- and immuno-sandwich magnetic assay for analysis of serum HER-2/neu protein overexpressed in several aggressive cancers. In the assay, we used a covalent hemin-guanine quadruplex (G4) complex as a novel O2-dependent electrocatalytic label that allowed 10 fM (aptamer-aptamer) and 1 fM (aptamer-antibody) detection of HER-2/neu in human serum. The O2 reactivity of the aptamer-conjugated label was detected at high-surface-area graphite electrodes displaying a high efficiency of O2 reduction electro-catalyzed by this DNAzyme. In contrast to the recognised H2O2 reactivity, the O2 reactivity of the covalent hemin/G4 complex depended only on ambient O2 present in solutions, and did not require adding such traditional reagents as hemin and H2O2, and solution de-aeration. Human serum albumin, urokinase plasminogen activator and thrombin did not interfere, and the assay was used for analysis of basal serum levels of HER-2/neu. Due to the simplicity and low cost, sandwich assays exploiting O2-linked electrocatalysis by the covalent hemin-G4 complexes represent a more advanced electrochemical ELISA platform for ultrasensitive and fast detection of low concentrations of proteins in complex biological matrices.

KW - Antibody

KW - Aptamer

KW - Covalent Hemin/G4 complex

KW - DNAzyme

KW - Electrochemical sandwich assay

KW - HER-2/neu

KW - Magnetic beads

KW - O electrocatalysis

KW - Limit of Detection

KW - Oxygen

KW - Hydrogen Peroxide/chemistry

KW - Humans

KW - Hemin/chemistry

KW - DNA, Catalytic/chemistry

KW - Magnetic Phenomena

KW - Biosensing Techniques

KW - Biological Assay

KW - G-Quadruplexes

KW - Aptamers, Nucleotide/chemistry

UR - http://www.scopus.com/inward/record.url?scp=85131460271&partnerID=8YFLogxK

U2 - 10.1016/j.aca.2022.340049

DO - 10.1016/j.aca.2022.340049

M3 - Journal article

C2 - 35715131

AN - SCOPUS:85131460271

SN - 0003-2670

VL - 1219

JO - Analytica Chimica Acta

JF - Analytica Chimica Acta

M1 - 340049

ER -

Covalent Hemin/G4 complex-linked sandwich bioassay on magnetic beads for femtomolar HER-2/neu detection in human serum via direct electrocatalytic reduction of oxygen

Abstract

Keywords

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