Creation of a DNA aptamer biosensor for the detection of heart failure - Couverture souple

Darnaud, Marion

 
9783659829239: Creation of a DNA aptamer biosensor for the detection of heart failure
Couverture souple
ISBN 10 : 3659829234 ISBN 13 : 9783659829239
Editeur : LAP LAMBERT Academic Publishing, 2016

Synopsis

Aptamers are small nucleic acid molecules, which are used to bind specific targets in ways similar to antibodies. They offer advantages in terms of cost and reproducibility of production, stability and capability for biochemical modifications. This work focused on discovering a new DNA aptamer biosensor for brain natriuretic peptide (BNP), a biomarker of the early stages of heart failure. A library of random DNA sequences was generated by the polymerase chain reaction and evaluated for specific and sensitive binding to BNP using an in vitro selection technique known as systematic evolution of ligands by exponential enrichment (SELEX). After 15 SELEX cycles using BNP immobilised on an affinity chromatography column, selected aptamers were subcloned and sequenced. The aptamer exhibiting the best specificity towards BNP was shown to adopt a G-quadruplex structure of the anti-parallel type. This aptamer was used to devise an enzyme-linked oligonucleotide assay using surface enhanced Raman spectroscopy allowing the detection of BNP down to the picomolar range. In addition, a quick colorimetric assay was developed on the concept of target-mediated aggregation of gold nanoparticles.

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Présentation de l'éditeur

Aptamers are small nucleic acid molecules, which are used to bind specific targets in ways similar to antibodies. They offer advantages in terms of cost and reproducibility of production, stability and capability for biochemical modifications. This work focused on discovering a new DNA aptamer biosensor for brain natriuretic peptide (BNP), a biomarker of the early stages of heart failure. A library of random DNA sequences was generated by the polymerase chain reaction and evaluated for specific and sensitive binding to BNP using an in vitro selection technique known as systematic evolution of ligands by exponential enrichment (SELEX). After 15 SELEX cycles using BNP immobilised on an affinity chromatography column, selected aptamers were subcloned and sequenced. The aptamer exhibiting the best specificity towards BNP was shown to adopt a G-quadruplex structure of the anti-parallel type. This aptamer was used to devise an enzyme-linked oligonucleotide assay using surface enhanced Raman spectroscopy allowing the detection of BNP down to the picomolar range. In addition, a quick colorimetric assay was developed on the concept of target-mediated aggregation of gold nanoparticles.

Biographie de l'auteur

Marion Darnaud is a R&D Biosciences Manager. She graduated with a PhD in Pure and Applied Chemistry in 2008 at the University of Strathclyde, Glasgow, UK. Her PhD work, under the supervision of Prof. Duncan Graham, was dedicated to the creation of a new DNA aptamer and the development of biosensor assays for the detection of heart failure.

Les informations fournies dans la section « A propos du livre » peuvent faire référence à une autre édition de ce titre.

Éditeur
LAP LAMBERT Academic Publishing
Date d'édition
2016
Langue
anglais
ISBN 10 
3659829234
ISBN 13 
9783659829239
Reliure
Broché
Nombre de pages
332
Coordonnées du fabricant
non disponible
Personne responsable
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