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Rapid Complexation of Aptamers by Their Specific Antidotes

Nucleic acid ligands, aptamers, harbor the unique characteristics of small molecules and antibodies. The specificity and high affinity of aptamers enable their binding to different targets, such as small molecules, proteins, or cells. Chemical modifications of aptamers allow increased bioavailabilit...

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Autores principales: Stoll, Heidi, Steinle, Heidrun, Wilhelm, Nadja, Hann, Ludmilla, Kunnakattu, Silju-John, Narita, Miwako, Schlensak, Christian, Wendel, Hans P., Avci-Adali, Meltem
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6152687/
https://www.ncbi.nlm.nih.gov/pubmed/28594360
http://dx.doi.org/10.3390/molecules22060954
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author Stoll, Heidi
Steinle, Heidrun
Wilhelm, Nadja
Hann, Ludmilla
Kunnakattu, Silju-John
Narita, Miwako
Schlensak, Christian
Wendel, Hans P.
Avci-Adali, Meltem
author_facet Stoll, Heidi
Steinle, Heidrun
Wilhelm, Nadja
Hann, Ludmilla
Kunnakattu, Silju-John
Narita, Miwako
Schlensak, Christian
Wendel, Hans P.
Avci-Adali, Meltem
author_sort Stoll, Heidi
collection PubMed
description Nucleic acid ligands, aptamers, harbor the unique characteristics of small molecules and antibodies. The specificity and high affinity of aptamers enable their binding to different targets, such as small molecules, proteins, or cells. Chemical modifications of aptamers allow increased bioavailability. A further great benefit of aptamers is the antidote (AD)-mediated controllability of their effect. In this study, the AD-mediated complexation and neutralization of the thrombin binding aptamer NU172 and Toll-like receptor 9 (TLR9) binding R10-60 aptamer were determined. Thereby, the required time for the generation of aptamer/AD-complexes was analyzed at 37 °C in human serum using gel electrophoresis. Afterwards, the blocking of aptamers’ effects was analyzed by determining the activated clotting time (ACT) in the case of the NU172 aptamer, or the expression of immune activation related genes IFN-1β, IL-6, CXCL-10, and IL-1β in the case of the R10-60 aptamer. Gel electrophoresis analyses demonstrated the rapid complexation of the NU172 and R10-60 aptamers by complementary AD binding after just 2 min of incubation in human serum. A rapid neutralization of anticoagulant activity of NU172 was also demonstrated in fresh human whole blood 5 min after addition of AD. Furthermore, the TLR9-mediated activation of PMDC05 cells was interrupted after the addition of the R10-60 AD. Using these two different aptamers, the rapid antagonizability of the aptamers was demonstrated in different environments; whole blood containing numerous proteins, cells, and different small molecules, serum, or cell culture media. Thus, nucleic acid ADs are promising molecules, which offer several possibilities for different in vivo applications, such as antagonizing aptamer-based drugs, immobilization, or delivery of oligonucleotides to defined locations.
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spelling pubmed-61526872018-11-13 Rapid Complexation of Aptamers by Their Specific Antidotes Stoll, Heidi Steinle, Heidrun Wilhelm, Nadja Hann, Ludmilla Kunnakattu, Silju-John Narita, Miwako Schlensak, Christian Wendel, Hans P. Avci-Adali, Meltem Molecules Article Nucleic acid ligands, aptamers, harbor the unique characteristics of small molecules and antibodies. The specificity and high affinity of aptamers enable their binding to different targets, such as small molecules, proteins, or cells. Chemical modifications of aptamers allow increased bioavailability. A further great benefit of aptamers is the antidote (AD)-mediated controllability of their effect. In this study, the AD-mediated complexation and neutralization of the thrombin binding aptamer NU172 and Toll-like receptor 9 (TLR9) binding R10-60 aptamer were determined. Thereby, the required time for the generation of aptamer/AD-complexes was analyzed at 37 °C in human serum using gel electrophoresis. Afterwards, the blocking of aptamers’ effects was analyzed by determining the activated clotting time (ACT) in the case of the NU172 aptamer, or the expression of immune activation related genes IFN-1β, IL-6, CXCL-10, and IL-1β in the case of the R10-60 aptamer. Gel electrophoresis analyses demonstrated the rapid complexation of the NU172 and R10-60 aptamers by complementary AD binding after just 2 min of incubation in human serum. A rapid neutralization of anticoagulant activity of NU172 was also demonstrated in fresh human whole blood 5 min after addition of AD. Furthermore, the TLR9-mediated activation of PMDC05 cells was interrupted after the addition of the R10-60 AD. Using these two different aptamers, the rapid antagonizability of the aptamers was demonstrated in different environments; whole blood containing numerous proteins, cells, and different small molecules, serum, or cell culture media. Thus, nucleic acid ADs are promising molecules, which offer several possibilities for different in vivo applications, such as antagonizing aptamer-based drugs, immobilization, or delivery of oligonucleotides to defined locations. MDPI 2017-06-08 /pmc/articles/PMC6152687/ /pubmed/28594360 http://dx.doi.org/10.3390/molecules22060954 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Stoll, Heidi
Steinle, Heidrun
Wilhelm, Nadja
Hann, Ludmilla
Kunnakattu, Silju-John
Narita, Miwako
Schlensak, Christian
Wendel, Hans P.
Avci-Adali, Meltem
Rapid Complexation of Aptamers by Their Specific Antidotes
title Rapid Complexation of Aptamers by Their Specific Antidotes
title_full Rapid Complexation of Aptamers by Their Specific Antidotes
title_fullStr Rapid Complexation of Aptamers by Their Specific Antidotes
title_full_unstemmed Rapid Complexation of Aptamers by Their Specific Antidotes
title_short Rapid Complexation of Aptamers by Their Specific Antidotes
title_sort rapid complexation of aptamers by their specific antidotes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6152687/
https://www.ncbi.nlm.nih.gov/pubmed/28594360
http://dx.doi.org/10.3390/molecules22060954
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