Diagnostic kit based on halloysite nanoclay-ionic liquid nanocomposite modified electrode for electrochemical determination of cancer biomarker

Esma Yildiz; Beste Yurdacan; Yasemin Erac; Arzum Erdem

doi:10.1016/j.talanta.2022.123854

Diagnostic kit based on halloysite nanoclay-ionic liquid nanocomposite modified electrode for electrochemical determination of cancer biomarker

Talanta. 2023 Jan 15:252:123854. doi: 10.1016/j.talanta.2022.123854. Epub 2022 Aug 19.

Authors

Esma Yildiz¹, Beste Yurdacan², Yasemin Erac², Arzum Erdem³

Affiliations

¹ The Institute of Natural and Applied Sciences, Biomedical Technologies Department, Ege University, Bornova, 35100, Izmir, Turkey; Faculty of Pharmacy, Analytical Chemistry Department, Ege University, Bornova, 35100, Izmir, Turkey.
² Faculty of Pharmacy, Department of Pharmacology, Ege University, Bornova, 35100, Izmir, Turkey.
³ The Institute of Natural and Applied Sciences, Biomedical Technologies Department, Ege University, Bornova, 35100, Izmir, Turkey; Faculty of Pharmacy, Analytical Chemistry Department, Ege University, Bornova, 35100, Izmir, Turkey. Electronic address: arzum.erdem@ege.edu.tr.

PMID: 36029681
DOI: 10.1016/j.talanta.2022.123854

Abstract

Nucleic acid hybridization is occurred between the selective single-stranded nucleic acid sequence and its target sequence, which is one of the essential procedure for electrochemical detection of nucleic acid. microRNA-21 (miRNA-21) is known as a biomarker in various cancers. The determination of miRNA-21 was attained through by hybridization of inosine substituted miRNA-21 specific DNA probe (P_inosine) with its target miRNA-21. In this study, the surface of pencil graphite electrode (PGE) was firstly modified with halloysite nanoclay-ionic liquid (HNT/IL) nanocomposite. The characterization of surface was performed by Scanning Electron Microscope (SEM) images and Energy Dispersive X-Ray Analysis (EDX) analysis, and the differences at surface modifications were also shown by electrochemical methods with electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). For sensitive and selective determination of miRNA-21, P_inosine and target miRNA concentration, immobilization and hybridization time were optimized by using HNT/IL modified PGE in combination with differential pulse voltammetry (DPV). The detection limit was achieved as 0.17 μg/mL (equals to 23.69 nM) in the linear range of 0.25-2 μg/mL miRNA-21. The selectivity of voltammetric method based on HNT/IL-PGE developed for miRNA-21 was examined in the presence of mismatch (MM) and non-complementary (NC) sequences. Because miRNA-21 is over-expressed in cancer cells, it has been tested in total RNA samples isolated from cancer cell line (breast cancer cell line, MCF-7). In the total RNA samples obtained from MCF-7, the detection limit was calculated as 0.28 μg/mL in the linear range of 1-4 μg/mL. Besides, the healthy cell line (human embryonic kidney cell line, HEK-293) was used as a control group and the results obtained by MCF-7 total RNA samples were compared to the results using HEK-293 total RNA samples in terms of miRNA-21 level.

Keywords: Cancer biomarker; Diagnostic kit; Electrochemical detection; Halloysite nanoclay; Ionic liquid; miRNA-21.

MeSH terms

Biomarkers, Tumor / genetics
Biosensing Techniques* / methods
Clay
DNA Probes / chemistry
Electrochemical Techniques / methods
Electrodes
Graphite* / chemistry
HEK293 Cells
Humans
Inosine
Ionic Liquids*
MicroRNAs* / analysis
Nanocomposites*
Neoplasms* / diagnosis

Substances

Biomarkers, Tumor
Clay
DNA Probes
Graphite
Inosine
Ionic Liquids
MicroRNAs