Details zur Publikation

Referenztyp Zeitschriften
DOI / URL Link
Titel (primär) Identification of the target binding site of ethanolamine-binding aptamers and its exploitation for ethanolamine detection
Autor Heilkenbrinker, A.; Reinemann, C.; Stoltenburg, R.; Walter, J.G.; Jochums, A.; Stahl, F.; Zimmermann, S.; Strehlitz, B.; Scheper, T.;
Journal / Serie Analytical Chemistry
Erscheinungsjahr 2015
Department BOOEK; UBZ;
Band/Volume 87
Heft 1
Sprache englisch;
POF III (gesamt) T34; T11;
Supplements https://pubs.acs.org/doi/suppl/10.1021/ac5034819/suppl_file/ac5034819_si_001.pdf
UFZ Querschnittsthemen 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.
ID 15541
dauerhafte UFZ-Verlinkung 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