Binding kinetics of human cellular prion detection by DNA aptamers immobilized on a conducting polypyrrole
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ORIGINAL PAPER
Binding kinetics of human cellular prion detection by DNA aptamers immobilized on a conducting polypyrrole A. Miodek & A. Poturnayová & M. Šnejdárková & T. Hianik & H. Korri-Youssoufi
Received: 29 August 2012 / Revised: 12 December 2012 / Accepted: 13 December 2012 / Published online: 15 January 2013 # Springer-Verlag Berlin Heidelberg 2013
Abstract We developed a biosensor based on the surface plasmon resonance (SPR) method for the study of the binding kinetics and detection of human cellular prions (PrPC) using DNA aptamers as bioreceptors. The biosensor was formed by immobilization of various biotinylated DNA aptamers on a surface of conducting polypyrrole modified by streptavidin. We demonstrated that PrPC interaction with DNA aptamers could be followed by measuring the variation of the resonance angle. This was studied using DNA aptamers of various configurations, including conventional single-stranded aptamers that contained a rigid double-stranded supporting part and aptamer dimers containing two binding sites. The kinetic constants determined by the SPR method suggest strong interaction of PrPC with various DNA aptamers depending on their configuration. SPR aptasensors have a high selectivity to PrPC and were regenerable by a brief wash in 0.1 M NaOH. The best limit of detection
Electronic supplementary material The online version of this article (doi:10.1007/s00216-012-6665-4) contains supplementary material, which is available to authorized users. A. Miodek : H. Korri-Youssoufi (*) CNRS UMR-8182, Institut de Chimie Moléculaire et des Matériaux d’Orsay, Equipe de Chimie Bioorganique et Bioinorganique, Univ Paris-Sud, Bâtiment 420, 91405 Orsay, France e-mail: [email protected] A. Poturnayová : M. Šnejdárková Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, 900 28 Ivanka pri Dunaji, Slovakia T. Hianik Faculty of Mathematics, Physics and Informatics, Comenius University, Mlynska dolina F1, 84248 Bratislava, Slovakia
(4 nM) has been achieved with this biosensor based on DNA aptamers with one binding site but containing a doublestranded supporting part. Keywords Prions . Aptamers . Kinetics . SPR . Polypyrrole
Introduction The diagnosis and treatment of neurogenerative diseases, such as Alzheimer’s or Creutzfeldt–Jakob, are priorities under consideration for health institutions. It is assumed that these diseases are caused by transformation of cellular prions (PrPC) into their infectious isoform PrPSc [1, 2]. PrPSc differs from PrPC in its insolubility, high content of β sheets, partial resistance to protease digestion, and tendency to form large aggregates [3]. Therefore, the development of a rapid and sensitive method for the detection of PrPSc is crucial for early diagnostics of these diseases. Especially useful would be a point-of-care assay that can be used even by physicians outside of specialized clinical laboratories [4]. So far, mostly the immunodiagnostic procedure [5] and postmortem histopathological identification of brain tissues were appli
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