Aptamers

Leader

Dr. Víctor Manuel González Muñoz

Personnel

Dra. Mª Elena Martín Palma

Marta García-Hernández

Contact

Dra. Mª Elena Martín Palma
m.elena.martin(ELIMINAR)@hrc.es 
+34- 91 336 81 73

  • Equipment
    • Real-time thermal cycler IQ5 (BioRad)
    • End-time thermal cycler (TEKNE)
    • Experion automatic electrophoresis system (BioRad)
    • AKTA purification system (GE Healthcare Life Sciences)
    • AKTA purification system (GE Healthcare Life Sciences)
    • MIniBis Pro image acquisition and documentation system (DNR)
    • Nanophotometer (IMPLEN)
    • Centrifuges, incubators, thermal blocks, horizontal and vertical electrophoresis systems and cold chamber.
  • Highlights

    Publications

    • Safety and Efficacy of ApTOLL in Patients With Ischemic Stroke Undergoing Endovascular Treatment: A Phase 1/2 Randomized Clinical Trial. Hernández-Jiménez M, Abad-Santos F, Cotgreave I, .... González VM, Moro MA, Garcia-Tornel A, Lizasoain I, Ribo M. JAMA Neurol. 2023 Jun 20:e231660. doi: 10.1001/jamaneurol.2023.1660..
    • First-in-human phase I clinical trial of a TLR4-binding DNA aptamer, ApTOLL: Safety and pharmacokinetics in healthy volunteers. Hernández-Jiménez M, Martín-Vílchez S, Ochoa D..., González VM, Lizasoain I, Abad-Santos F. Mol Ther Nucleic Acids. 2022 Mar 9;28:124-135. doi: 10.1016/j.omtn.2022.03.005
    • Selection and characterization of DNA aptamers for highly selective recognition of the major allergen of olive pollen Ole e 1. Vergara-Barberán M, Lerma-García MJ, Simó-Alfonso EF, García-Hernández M, Martín ME, García-Sacristán A, González VM, Herrero-Martínez JM. Anal Chim Acta. 2022 Feb 1;1192:339334. doi: 10.1016/j.aca.2021.339334.
    • FPR2 DNA Aptamers for Targeted Therapy of Wound Repair. de Arriba MDC, Fernández G, Chacón-Solano E, Mataix M, Martínez-Santamaría L, Illera N, Carrión-Marchante R, Martín ME, Larcher F, González VM, Del Río M, Carretero M. J Invest Dermatol. 2022 Aug;142(8):2238-2248.e8. doi: 10.1016/j.jid.2021.12.026.
    • Development of an antigen Enzyme-Linked AptaSorbent Assay (ELASA) for the detection of swine influenza virus in field samples. Aira C, Klett-Mingo JI, Ruiz T, Garcia-Sacristán A, Martín-Valls GE, Mateu E, Gómez-Laguna J, Rueda P, González VM, Rodríguez MJ, López L. Anal Chim Acta. 2021 Oct 9;1181:338933. doi: 10.1016/j.aca.2021.338933.
    • The Relevance of the SH2 Domain for c-Src Functionality in Triple-Negative Breast Cancer Cells. Mayoral-Varo V, Sánchez-Bailón MP, Calcabrini A, García-Hernández M, Frezza V, Martín ME, González VM, Martín-Pérez J. Cancers (Basel). 2021 Jan 26;13(3):462. doi: 10.3390/cancers13030462.
    • Novel DNA Aptamers Against CCL21 Protein: Characterization and Biomedical Applications for Targeted Drug Delivery to T Cell-Rich Zones. Chonco L, Fernández G, Kalhapure R, Hernáiz MJ, García-Oliva C, Gonzalez VM, Martín ME, Govender T, Parboosing R. Nucleic Acid Ther. 2018 Aug;28(4):242-251. doi: 10.1089/nat.2017.0689

    Projects

    • Contract/Project Title: Selection and identification of aptamers against Xylella fastidiosa. Type of contract: Contract. Funding Company/Administration: Institute of Sustainable Agriculture-CSIC (2020-2023)
    • Contract/Project Title: Selection and Identification of Aptamers against a Protein of Botrytis cinerea. Type of contract: Contract. Funding Company/Administration: Dept. of Microbiology. Faculty of Sciences. University of Málaga (2020-2023)
    • Contract/Project Title: Selection of aptamers against cardiomyocytes. Type of contract: Contract. Funding Company/Administration: Universidad de Zaragoza (2022-2023)
    • Contract/Project Title: Selection of Aptamers Capable of Crossing the Myocardial Capillary Barrier. Type of contract: Contract. Funding Company/Administration: University of Zaragoza (2023)
    • Title of Contract/Project: Selection of aptamers against a protein of the outer membrane of spermatozoa. Type of contract: Contract. Funding Company/Administration: Department of Physiology. Faculty of Medicine and Dentistry. UPV (2020-2023)
    • Title of contract/project: Analysis of aptamer populations sequenced by NGS. Type of contract: Contract. Funding Company/Administration: Universidad de la República. Montevideo. Uruguay (2023)
    • Contract/Project Title: Preclinical studies of the apMNKQ2 aptamer. Type of contract: Contract. Funding Company/Administration: Aptus Biotech (2021-2023)
    • Contract/Project Title: Stability of aptamers in vitro. Type of contract: Contract. Funding Company/Administration: AptaTargets (2021-2023)
    • Contract/Project Title: Toxicity of aptamers in vitro. Type of contract: Contract. Funding Company/Administration: AptaTargets (2021-2023)
    • Contract/Project Title: Studies of aptamer binding to plasma proteins. Type of contract: Contract. Funding Company/Administration: AptaTargets (2021-2023)
    • Contract/Project Title: Studies of the Effect of Aptamers on the Formation of Nanospheres. Type of contract: Contract. Funding Company/Administration: AptaTargets (2021-2023)
    • Contract/Project Title: Selection of aptamers against HIV peptides. Type of contract: Contract. Funding Company/Administration: Africa Holguin /IRYCIS) (2020-2022)
    • Contract/Project Title: Selection of aptamers against olfactory receptors. Type of contract: Contract. Funding Company/Administration: Department of Basic Research. CIEMAT (2016-2021)
    • Contract/Project Title: Selection and characterization of aptamers against SH2. Type of contract: Contract. Funding Company/Administration: Instituto de Investigaciones Biomédicas Alberto Sols. CIEMAT (2020-2021)

    Patents

    • TLR-4-SPECIFIC APTAMERS AND THEIR USESP201430955; PCT/EP2015/064277.
    • MAFG AS A POTENTIAL THERAPEUTIC TARGET TO RESTORE CHEMOSENSITIVITY IN PLATINUM-RESISTANT CANCER CELLS. EP17382610 with PCTEP2018/06815 and extension in USA 16/647,368 and Europe 18 734 266.2
    • APTAMERS AGAINST THE HEPATITIS C VIRUS CORE PROTEIN. EP21382710.8
    • APTAMERS AND THEIR USE IN THE TREATMENT OF CANCER. EP 18 382 888.8 with PCT/EP2019/083547
    • FPR2 RECEPTOR AGONIST APTAMERS AND USES THEREOF. P201930524 with PCT/ES2020/070378
  • Service portfolio and prices

    **Consult fees with the unit

    Selection of target-specific aptamer populations

    Populations of aptamers against a target shall be selected using the appropriate SELEX methodology. Up to 12 rounds of selection shall be performed. The enrichment of each of the populations obtained after successive rounds of selection shall be measured. The population enriched in aptamers recognising the target shall be provided.


    NGS identification and in silico structural characterisation of aptamers.

    The aptamer-enriched population will be sequenced against the target, the secondary structure of the identified aptamers will be analysed and up to 4 representative aptamers will be synthesised.

    Confirmation of interaction and SPR affinity study (4 aptamers)

    The interaction of the selected aptamers will be confirmed and the affinity of the chemically synthesised aptamers will be studied by SPR. The sequence of the aptamers showing the highest affinity for the target will be provided.

    Optimisation of aptamers

    The structure of the aptamers will be studied and modified aptamers will be designed by eliminating sequences that do not alter the active structure of the aptamer.

    In vitro stability: 1. In human, rat and monkey plasma (NHP); 2. Stability against nucleases (λ-exonuclease and DNAse I)

    The interaction of the selected aptamers will be confirmed and the affinity of the chemically synthesised aptamers will be studied by SPR. The sequence of the aptamers showing the highest affinity for the target will be provided.

    In vitro toxicity: assessment of aptamer cytotoxicity by MTT and LDH quantification.

    Toxicity of samples in cell cultures shall be assessed by MTT and LDH quantification.

    Functional assays

    Studies to determine the agonist or antagonist activity of the aptamers shall be performed using appropriate models.

    Technical advice and support

    Advice related to aptamer technology. Development of new protocols. Training and implementation of specific techniques. Technical and methodological support.