The characterization and validation of 17β-estradiol binding aptamers.
The rapid and sensitive detection of small molecules is garnering increasing importance, and aptamers show great promise in replacing expensive, elaborate detection platforms exploiting chromatographic separation or antibody-based assays. The characterization of aptamer interaction with small molecule targets is not facile, and there is a mature need for a rapid, high-throughput technique for the analysis of aptamer-small molecule kinetics and affinity. In this work we present methodologies for the evaluation of aptamer-small molecule interactions, using the aptamers reported against the steroid 17β-estradiol as a model system. Microscale thermophoresis, apta-PCR affinity assay and surface plasmon resonance were explored to evaluate the reported aptamers' binding properties in terms of affinity and specificity, and were demonstrated to be successfully applied to the analysis of aptamer-small molecule interactions.
Interfibio, Nanobiotechnology & Bioanalysis Group, Departament d'Enginyeria Química, Universitat Rovira i Virgili, Avinguda Països Catalans 26, Tarragona 43007, Spain.
Markéta
M
Svobodová
Interfibio, Nanobiotechnology & Bioanalysis Group, Departament d'Enginyeria Química, Universitat Rovira i Virgili, Avinguda Països Catalans 26, Tarragona 43007, Spain. Electronic address: vasoula.skouridou@urv.cat.
Vasso
V
Skouridou
Interfibio, Nanobiotechnology & Bioanalysis Group, Departament d'Enginyeria Química, Universitat Rovira i Virgili, Avinguda Països Catalans 26, Tarragona 43007, Spain.
Mary Luz
ML
Botero
Interfibio, Nanobiotechnology & Bioanalysis Group, Departament d'Enginyeria Química, Universitat Rovira i Virgili, Avinguda Països Catalans 26, Tarragona 43007, Spain.
Miriam
M
Jauset-Rubio
2bind GmbH, Josef Engert Strasse 13, Regensburg 93053, Germany.
Thomas
T
Schubert
Department of Chemistry, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia.
Abdulaziz S
AS
Bashammakh
Department of Chemistry, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia.
Mohammad S
MS
El-Shahawi
Department of Chemistry, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia.
Abdulrahman O
AO
Alyoubi
Interfibio, Nanobiotechnology & Bioanalysis Group, Departament d'Enginyeria Química, Universitat Rovira i Virgili, Avinguda Països Catalans 26, Tarragona 43007, Spain; Institució Catalana de Recerca i Estudis Avançats (ICREA), Passeig Lluís Companys 23, Barcelona 08010, Spain. Electronic address: ciara.osullivan@urv.cat.
Ciara K
CK
O'Sullivan
D052157:Aptamers, Nucleotide / Q000378:metabolism*; D001665:Binding Sites; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000378:metabolism*; D054856:G-Quadruplexes; D006801:Humans; D007368:Interferometry; D007700:Kinetics; D008024:Ligands; D008280:Magnetics; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction; D011374:Progesterone / Q000737:chemistry; D011485:Protein Binding; D052156:SELEX Aptamer Technique / Q000379:methods*; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance
D052157:Aptamers, Nucleotide; D008024:Ligands; D011374:Progesterone; D004958:Estradiol
17β-estradiol; Apta-PCR affinity assay (APAA); Aptamers; Microscale thermophoresis (MST); Small molecule; Surface Plasmon Resonance (SPR)