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Development of an automated in vitro selection protocol to obtain RNA-based aptamers: identification of a biostable substance P antagonist

We have developed an automated SELEX (Systematic Evolution of Ligands by EXponential Enrichment) process that allows the execution of in vitro selection cycles without any direct manual intervention steps. The automated selection protocol is designed to provide for high flexibility and versatility i...

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Detalles Bibliográficos
Autores principales: Eulberg, Dirk, Buchner, Klaus, Maasch, Christian, Klussmann, Sven
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2005
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC552970/
https://www.ncbi.nlm.nih.gov/pubmed/15745995
http://dx.doi.org/10.1093/nar/gni044
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author Eulberg, Dirk
Buchner, Klaus
Maasch, Christian
Klussmann, Sven
author_facet Eulberg, Dirk
Buchner, Klaus
Maasch, Christian
Klussmann, Sven
author_sort Eulberg, Dirk
collection PubMed
description We have developed an automated SELEX (Systematic Evolution of Ligands by EXponential Enrichment) process that allows the execution of in vitro selection cycles without any direct manual intervention steps. The automated selection protocol is designed to provide for high flexibility and versatility in terms of choice of buffers and reagents as well as stringency of selection conditions. Employing the automated SELEX process, we have identified RNA aptamers to the mirror-image configuration (d-peptide) of substance P. The peptide substance P belongs to the tachykinin family and exerts various biologically important functions, such as peripheral vasodilation, smooth muscle contraction and pain transmission. The aptamer that was identified most frequently was truncated to the 44mer SUP-A-004. The mirror-image configuration of SUP-A-004, the so-called Spiegelmer, has been shown to bind to naturally occurring l-substance P displaying a K(d) of 40 nM and to inhibit (IC(50) of 45 nM) l-substance P-mediated Ca(2+) release in a cell culture assay.
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spelling pubmed-5529702005-03-10 Development of an automated in vitro selection protocol to obtain RNA-based aptamers: identification of a biostable substance P antagonist Eulberg, Dirk Buchner, Klaus Maasch, Christian Klussmann, Sven Nucleic Acids Res Methods Online We have developed an automated SELEX (Systematic Evolution of Ligands by EXponential Enrichment) process that allows the execution of in vitro selection cycles without any direct manual intervention steps. The automated selection protocol is designed to provide for high flexibility and versatility in terms of choice of buffers and reagents as well as stringency of selection conditions. Employing the automated SELEX process, we have identified RNA aptamers to the mirror-image configuration (d-peptide) of substance P. The peptide substance P belongs to the tachykinin family and exerts various biologically important functions, such as peripheral vasodilation, smooth muscle contraction and pain transmission. The aptamer that was identified most frequently was truncated to the 44mer SUP-A-004. The mirror-image configuration of SUP-A-004, the so-called Spiegelmer, has been shown to bind to naturally occurring l-substance P displaying a K(d) of 40 nM and to inhibit (IC(50) of 45 nM) l-substance P-mediated Ca(2+) release in a cell culture assay. Oxford University Press 2005 2005-03-03 /pmc/articles/PMC552970/ /pubmed/15745995 http://dx.doi.org/10.1093/nar/gni044 Text en © The Author 2005. Published by Oxford University Press. All rights reserved
spellingShingle Methods Online
Eulberg, Dirk
Buchner, Klaus
Maasch, Christian
Klussmann, Sven
Development of an automated in vitro selection protocol to obtain RNA-based aptamers: identification of a biostable substance P antagonist
title Development of an automated in vitro selection protocol to obtain RNA-based aptamers: identification of a biostable substance P antagonist
title_full Development of an automated in vitro selection protocol to obtain RNA-based aptamers: identification of a biostable substance P antagonist
title_fullStr Development of an automated in vitro selection protocol to obtain RNA-based aptamers: identification of a biostable substance P antagonist
title_full_unstemmed Development of an automated in vitro selection protocol to obtain RNA-based aptamers: identification of a biostable substance P antagonist
title_short Development of an automated in vitro selection protocol to obtain RNA-based aptamers: identification of a biostable substance P antagonist
title_sort development of an automated in vitro selection protocol to obtain rna-based aptamers: identification of a biostable substance p antagonist
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC552970/
https://www.ncbi.nlm.nih.gov/pubmed/15745995
http://dx.doi.org/10.1093/nar/gni044
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