Author, as appears in the article.: Jauset-Rubio, Miriam; Luz Botero, Mary; Skouridou, Vasso; Betul Aktas, Gulsen; Svobodova, Marketa; Bashammakh, Abdulaziz S.; El-Shahawi, Mohammad S.; Alyoubi, Abdulrahman O.; O'Sullivan, Ciara K.;

Department: Enginyeria Química

URV's Author/s: AKTAS, GÜLSEN BETÜL / BOTERO GALLEGO, MARY LUZ / Jauset Rubio, Miriam / O'SULLIVAN, CIARA KATHLEEN / Skouridou, Vasoula

Keywords: Progesterone Ligands In-vitro selection Evolution Dna aptamer

Abstract: Aptamers are well-established biorecognition molecules used in a wide variety of applications for the detection of their respective targets. However, individual SELEX processes typically performed for the identification of aptamers for each target can be quite time-consuming, labor-intensive, and costly. An alternative strategy is proposed herein for the simultaneous identification of different aptamers binding distinct but structurally similar targets in one single selection. This one-pot SELEX approach, using the steroids estradiol, progesterone, and testosterone as model targets, was achieved by combining the benefits of counter-SELEX with the power of next-generation sequencing and bioinformatics analysis. The pools from the last stage of the selection were compared in order to discover sequences with preferential abundance in only one of the pools. This led to the identification of aptamer candidates with potential specificity to a single steroid target. Binding studies demonstrated the high affinity of each selected aptamer for its respective target, and low nanomolar range dissociation constants calculated were similar to those previously reported for steroid-binding aptamers selected using traditional SELEX approaches. Finally, the selected aptamers were exploited in microtiter plate assays, achieving nanomolar limits of detection, while the specificity of these aptamers was also demonstrated. Overall, the one-pot SELEX strategy led to the discovery of aptamers for three different steroid targets in one single selection without compromising their affinity or specificity, demonstrating the power of this approach of aptamer discovery for the simultaneous selection of aptamers against multiple targets.

Thematic Areas: Química Interdisciplinar General chemistry General chemical engineering Engenharias ii Ciências agrárias i Chemistry, multidisciplinary Chemistry (miscellaneous) Chemistry (all) Chemical engineering (miscellaneous) Chemical engineering (all)

licence for use: https://creativecommons.org/licenses/by/3.0/es/

ISSN: 24701343

Author's mail: miriam.jauset@urv.cat vasoula.skouridou@urv.cat

Author identifier: 0000-0002-9943-6132 0000-0002-9712-5429

Record's date: 2023-02-18

Papper version: info:eu-repo/semantics/publishedVersion

Link to the original source: https://pubs.acs.org/doi/abs/10.1021/acsomega.9b02412

Licence document URL: http://repositori.urv.cat/ca/proteccio-de-dades/

Papper original source: Acs Omega. 4 (23): 20188-20196

APA: Jauset-Rubio, Miriam; Luz Botero, Mary; Skouridou, Vasso; Betul Aktas, Gulsen; Svobodova, Marketa; Bashammakh, Abdulaziz S.; El-Shahawi, Mohammad S.; (2019). One-Pot SELEX: Identification of Specific Aptamers against Diverse Steroid Targets in One Selection. Acs Omega, 4(23), 20188-20196. DOI: 10.1021/acsomega.9b02412

Article's DOI: 10.1021/acsomega.9b02412

Entity: Universitat Rovira i Virgili

Journal publication year: 2019

Publication Type: Journal Publications