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2007-08-15Zeitschriftenartikel DOI: 10.1186/1472-6750-7-48
One-step selection of Vaccinia virus-binding DNA aptamers by MonoLEX
dc.contributor.authorNitsche, Andreas
dc.contributor.authorKurth, Andreas
dc.contributor.authorDunkhorst, Anna
dc.contributor.authorPänke, Oliver
dc.contributor.authorSielaff, Hendrik
dc.contributor.authorJunge, Wolfgang
dc.contributor.authorMuth, Doreen
dc.contributor.authorScheller, Frieder
dc.contributor.authorStöcklein, Walter
dc.contributor.authorDahmen, Claudia
dc.contributor.authorPauli, Georg
dc.contributor.authorKage, Andreas
dc.date.accessioned2018-05-07T13:47:00Z
dc.date.available2018-05-07T13:47:00Z
dc.date.created2010-03-23
dc.date.issued2007-08-15none
dc.identifier.otherhttp://edoc.rki.de/oa/articles/reFsqEkhyzshs/PDF/24PZE1DHhX5f.pdf
dc.identifier.urihttp://edoc.rki.de/176904/609
dc.description.abstractBackground: As a new class of therapeutic and diagnostic reagents, more than fifteen years ago RNA and DNA aptamers were identified as binding molecules to numerous small compounds, proteins and rarely even to complete pathogen particles. Most aptamers were isolated from complex libraries of synthetic nucleic acids by a process termed SELEX based on several selection and amplification steps. Here we report the application of a new one-step selection method (MonoLEX) to acquire high-affinity DNA aptamers binding Vaccinia virus used as a model organism for complex target structures. Results: The selection against complete Vaccinia virus particles resulted in a 64-base DNA aptamer specifically binding to orthopoxviruses as validated by dot blot analysis, Surface Plasmon Resonance, Fluorescence Correlation Spectroscopy and real-time PCR, following an aptamer blotting assay. The same oligonucleotide showed the ability to inhibit in vitro infection of Vaccinia virus and other orthopoxviruses in a concentration-dependent manner. Conclusion: The MonoLEX method is a straightforward procedure as demonstrated here for the identification of a high-affinity DNA aptamer binding Vaccinia virus. MonoLEX comprises a single affinity chromatography step, followed by subsequent physical segmentation of the affinity resin and a single final PCR amplification step of bound aptamers. Therefore, this procedure improves the selection of high affinity aptamers by reducing the competition between aptamers of different affinities during the PCR step, indicating an advantage for the single-round MonoLEX method.ger
dc.language.isoeng
dc.publisherRobert Koch-Institut
dc.subjectDNAeng
dc.subjectViral/geneticseng
dc.subjectViral/isolation & purificationeng
dc.subjectPolymerase Chain Reaction/methodseng
dc.subjectChromatographyeng
dc.subjectAptamerseng
dc.subjectNucleotide/geneticseng
dc.subjectNucleotide/isolation & purificationeng
dc.subjectAffinity/methodseng
dc.subjectGene Targeting/methodseng
dc.subjectVaccinia virus/geneticseng
dc.subject.ddc610 Medizin
dc.titleOne-step selection of Vaccinia virus-binding DNA aptamers by MonoLEX
dc.typeperiodicalPart
dc.identifier.urnurn:nbn:de:0257-1006197
dc.identifier.doi10.1186/1472-6750-7-48
dc.identifier.doihttp://dx.doi.org/10.25646/534
local.edoc.container-titleBMC Biotechnology
local.edoc.fp-subtypeArtikel
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-urlhttp://www.biomedcentral.com/1472-6750/7/48/abstract/
local.edoc.container-publisher-nameBioMedCentral
local.edoc.container-volume7
local.edoc.container-issue48
local.edoc.container-year2007

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