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A novel zinc finger protein-based amperometric biosensor for miRNA determination

The result's identifiers

  • Result code in IS VaVaI

    <a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00209805%3A_____%2F20%3A00078274" target="_blank" >RIV/00209805:_____/20:00078274 - isvavai.cz</a>

  • Result on the web

    <a href="https://link.springer.com/article/10.1007%2Fs00216-019-02219-w" target="_blank" >https://link.springer.com/article/10.1007%2Fs00216-019-02219-w</a>

  • DOI - Digital Object Identifier

    <a href="http://dx.doi.org/10.1007/s00216-019-02219-w" target="_blank" >10.1007/s00216-019-02219-w</a>

Alternative languages

  • Result language

    angličtina

  • Original language name

    A novel zinc finger protein-based amperometric biosensor for miRNA determination

  • Original language description

    This paper reports a simple electrochemical strategy for the determination of microRNAs (miRNAs) using a commercial His-Tag-Zinc finger protein (His-Tag-ZFP) that binds preferably (but non-sequence specifically) RNA hybrids over ssRNAs, ssDNAs, and dsDNAs. The strategy involves the use of magnetic beads (His-Tag-Isolation-MBs) as solid support to capture the conjugate formed in homogenous solution between His-Tag-ZFP and the dsRNA homohybrid formed between the target miRNA (miR-21 selected as a model) and a biotinylated synthetic complementary RNA detector probe (b-RNA-Dp) further conjugated with a streptavidin-horseradish peroxidase (Strep-HRP) conjugate. The electrochemical detection is carried out by amperometry at disposable screen-printed carbon electrodes (SPCEs) (- 0.20 V vs Ag pseudo-reference electrode) upon magnetic capture of the resultant magnetic bioconjugates and H2O2 addition in the presence of hydroquinone (HQ). The as-prepared biosensor exhibits a dynamic concentration range from 3.0 to 100 nM and a detection limit (LOD) of 0.91 nM for miR-21 in just ~ 2 h. An acceptable discrimination was achieved between the target miRNA and other non-target nucleic acids (ssDNA, dsDNA, ssRNA, DNA-RNA, miR-122, miR-205, and single central- or terminal-base mismatched sequences). The biosensor was applied to the analysis of miR-21 from total RNA (RNAt) extracted from epithelial non-tumorigenic and adenocarcinoma breast cells without target amplification, pre-concentration, or reverse transcription steps. The versatility of the methodology due to the ZFP&apos;s non-sequence-specific binding behavior makes it easily extendable to determine any target RNA only by modifying the biotinylated detector probe.

  • Czech name

  • Czech description

Classification

  • Type

    J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database

  • CEP classification

  • OECD FORD branch

    10405 - Electrochemistry (dry cells, batteries, fuel cells, corrosion metals, electrolysis)

Result continuities

  • Project

    Result was created during the realization of more than one project. More information in the Projects tab.

  • Continuities

    P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Others

  • Publication year

    2020

  • Confidentiality

    S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Data specific for result type

  • Name of the periodical

    Analytical and bioanalytical chemistry

  • ISSN

    1618-2642

  • e-ISSN

  • Volume of the periodical

    412

  • Issue of the periodical within the volume

    21

  • Country of publishing house

    DE - GERMANY

  • Number of pages

    11

  • Pages from-to

    5031-5041

  • UT code for WoS article

    000545670200005

  • EID of the result in the Scopus database

    2-s2.0-85075203278