Publication Details

Category Text Publication
Reference Category Journals
DOI 10.1021/ac5034819
Title (Primary) Identification of the target binding site of ethanolamine-binding aptamers and its exploitation for ethanolamine detection
Author Heilkenbrinker, A.; Reinemann, C.; Stoltenburg, R.; Walter, J.G.; Jochums, A.; Stahl, F.; Zimmermann, S.; Strehlitz, B.; Scheper, T.
Source Titel Analytical Chemistry
Year 2015
Department BOOEK; UBZ
Volume 87
Issue 1
Page From 677
Page To 685
Language englisch
Supplements https://pubs.acs.org/doi/suppl/10.1021/ac5034819/suppl_file/ac5034819_si_001.pdf
UFZ wide themes RU1
Abstract Aptamers are promising recognition elements for sensitive and specific detection of small molecules. We have previously selected ssDNA aptamers for ethanolamine, one of the smallest aptamer targets so far. The work presented here focuses on the determination of the binding region within the aptamer structure and its exploitation for the development of an aptamer-based assay for detection of ethanolamine. Sequence analysis of the aptamers resulted in the identification of a G-rich consensus sequence which was able to fold in a typical two- or three- layered G-quartet structure. Experiments with stepwise truncated variants of the aptamers revealed that the consensus sequence is responsible and sufficient for binding to the target. Based on the knowledge of the aptamers binding site, we developed an aptamer-based microarray assay relying on competition between ethanolamine and an oligonucleotide complementary to the consensus sequence. Competitive binding of ethanolamine and fluorescently labeled complementary oligonucleotides resulted in fluorescence intensities dependent on ethanolamine concentration with a limit of detection of 10 pM. This method enables detection of small molecules without any labeling of analytes. The competitive assay could potentially be transferred to other aptamers and thus provides a promising system for aptamer-based detection of diverse small molecules.
Persistent UFZ Identifier https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=15541
Heilkenbrinker, A., Reinemann, C., Stoltenburg, R., Walter, J.G., Jochums, A., Stahl, F., Zimmermann, S., Strehlitz, B., Scheper, T. (2015):
Identification of the target binding site of ethanolamine-binding aptamers and its exploitation for ethanolamine detection
Anal. Chem. 87 (1), 677 - 685 10.1021/ac5034819