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Anticipating metastasis through electrochemical immunosensing of tumor hypoxia biomarkers

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Abstract

Metastasis is responsible for about 90% of cancer-associated deaths. In the context of solid tumors, the low oxygen concentration in the tumor microenvironment (hypoxia) is one of the key factors contributing to metastasis. Tumor cells adapt to these conditions by overexpressing certain proteins such as programmed death ligand 1 (PD-L1) and hypoxia-inducible factor 1 alpha (HIF-1α). However, the determination of these tumor hypoxia markers that can be used to follow-up tumor progression and improve the efficiency of therapies has been scarcely addressed using electrochemical biosensors. In this work, we report the first electrochemical bioplatform for the determination of PD-L1 as well as the first one allowing its simultaneous determination with HIF-1α. The target proteins were captured and enzymatically labeled on magnetic microbeads and amperometric detection was undertaken on the surface of screen-printed dual carbon electrodes using the hydrogen peroxide/peroxidase/hydroquinone system. Sandwich immunoassays were implemented for both the HIF-1α and PD-L1 sensors and the analytical characteristics were evaluated providing LOD values of 86 and 279 pg mL−1 for the amperometric determination of PD-L1 and HIF-1α standards, respectively. The developed electrochemical immunoplatforms are competitive versus the only electrochemical immunosensor reported for the determination of HIF-1α and the “gold standard” ELISA methodology for the single determination of both proteins in terms of assay time, compatibility with the simultaneous determination of both proteins making their use suitable for untrained users at the point of attention. The dual amperometric immunosensor was applied to the simultaneous determination of HIF-1α and PD-L1 in cancer cell lysates. The analyses lasted only 2 h and just 0.5 μg of the sample was required.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

The financial support of PID2019-103899RB-I00 (Spanish Ministerio de Ciencia e Innovación), RTI2018-096135-B-I00 (Spanish Ministerio de Ciencia, Innovación y Universidades) Research Projects, PI17CIII/00045 and PI20CIII/00019 Grants from the AES-ISCIII Program co-founded by FEDER funds, and the TRANSNANOAVANSENS-CM Program from the Comunidad de Madrid (Grant S2018/NMT-4349) are gratefully acknowledged. C.M.-S.M. acknowledges a predoctoral contract from Universidad Complutense de Madrid. A.M-C. is supported by a FPU predoctoral contract supported by the Spanish Ministerio de Educación, Cultura y Deporte.

Funding

Spanish Ministerio de Ciencia e Innovación (PID2019-103899RB-I00), Spanish Ministerio de Ciencia, Innovación y Universidades (RTI2018–096135-B-I00), AES-ISCIII Program co-founded by FEDER funds (PI17CIII/00045 and PI20CIII/00019 Grants), TRANSNANOAVANSENS-CM Program from the Comunidad de Madrid (Grant S2018/NMT-4349).

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Authors and Affiliations

  1. Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, 28040, Madrid, Spain

    Cristina Muñoz-San Martín, Maria Gamella, María Pedrero, Víctor Pérez-Ginés, José M. Pingarrón & Susana Campuzano

  2. UFIEC, Instituto de Salud Carlos III, 28220, Majadahonda, Madrid, Spain

    Ana Montero-Calle & Rodrigo Barderas

  3. Unitat de Recerca Biomèdica, Hospital Universitari Sant Joan, Institut d’Investigació Sanitària Pere Virgili, Universitat Rovirai Virgili, 43204, Reus, Spain

    Jordi Camps & Meritxell Arenas

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  1. Cristina Muñoz-San Martín
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Contributions

Cristina Muñoz-San Martín: conceptualization, investigation, writing, review, and editing—original draft. Maria Gamella: conceptualization, investigation, writing, review, and editing—original draft. María Pedrero: conceptualization, investigation, writing, review, and editing—original draft. Ana Montero-Calle: investigation, resources, review, and editing—original draft. Víctor Pérez-Ginés: investigation. Rodrigo Barderas: conceptualization, supervision, resources, review, and editing—original draft and funding acquisition. José M. Pingarrón: Review and editing—original draft. Susana Campuzano: conceptualization, supervision, writing, review, and editing—original draft and funding acquisition.

Corresponding author

Correspondence to Susana Campuzano.

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Muñoz-San Martín, C., Gamella, M., Pedrero, M. et al. Anticipating metastasis through electrochemical immunosensing of tumor hypoxia biomarkers. Anal Bioanal Chem 414, 399–412 (2022). https://doi.org/10.1007/s00216-021-03240-8

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