Articles producció científica> Enginyeria Química

One-Pot SELEX: Identification of Specific Aptamers against Diverse Steroid Targets in One Selection

  • Dades identificatives

    Identificador: imarina:6012540
    Autors:
    Jauset-Rubio, MiriamLuz Botero, MarySkouridou, VassoBetul Aktas, GulsenSvobodova, MarketaBashammakh, Abdulaziz SEl-Shahawi, Mohammad SAlyoubi, Abdulrahman OO'Sullivan, Ciara K
    Resum:
    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.
  • Altres:

    Autor segons l'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
    Departament: Enginyeria Química
    Autor/s de la URV: AKTAS, GÜLSEN BETÜL / BOTERO GALLEGO, MARY LUZ / Jauset Rubio, Miriam / O'SULLIVAN, CIARA KATHLEEN / Skouridou, Vasoula / SVOBODOVÁ, MARKÉTA
    Paraules clau: Progesterone Ligands In-vitro selection Evolution Dna aptamer
    Resum: 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.
    Àrees temàtiques: Química Interdisciplinar General chemistry General chemical engineering Engenharias ii Ciências agrárias i Chemistry, multidisciplinary Chemistry (miscellaneous) Chemistry (all) Chemical engineering (miscellaneous) Chemical engineering (all)
    Accès a la llicència d'ús: https://creativecommons.org/licenses/by/3.0/es/
    ISSN: 24701343
    Adreça de correu electrònic de l'autor: vasoula.skouridou@urv.cat miriam.jauset@urv.cat
    Identificador de l'autor: 0000-0002-9712-5429 0000-0002-9943-6132
    Data d'alta del registre: 2024-10-12
    Versió de l'article dipositat: info:eu-repo/semantics/publishedVersion
    URL Document de llicència: https://repositori.urv.cat/ca/proteccio-de-dades/
    Referència a l'article segons font original: Acs Omega. 4 (23): 20188-20196
    Referència de l'ítem segons les normes APA: Jauset-Rubio, Miriam; Luz Botero, Mary; Skouridou, Vasso; Betul Aktas, Gulsen; Svobodova, Marketa; Bashammakh, Abdulaziz S; El-Shahawi, Mohammad S; Al (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
    Entitat: Universitat Rovira i Virgili
    Any de publicació de la revista: 2019
    Tipus de publicació: Journal Publications
  • Paraules clau:

    Chemical Engineering (Miscellaneous),Chemistry (Miscellaneous),Chemistry, Multidisciplinary
    Progesterone
    Ligands
    In-vitro selection
    Evolution
    Dna aptamer
    Química
    Interdisciplinar
    General chemistry
    General chemical engineering
    Engenharias ii
    Ciências agrárias i
    Chemistry, multidisciplinary
    Chemistry (miscellaneous)
    Chemistry (all)
    Chemical engineering (miscellaneous)
    Chemical engineering (all)
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