Anticipating metastasis through electrochemical immunosensing of tumor hypoxia biomarkers
C. Muñoz-San Martín, M. Gamella, M. Pedrero, A. Montero-Calle, V. Pérez-Ginés, J. Camps, M. Arenas, R. Barderas, J.M. Pingarrón, S. Campuzano
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-1alpha). 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-1alpha.
Electrochemical LAMP-based assay for detection of RNA biomarkers in prostate cancer
L. Moranova, M. Stanik, R. Hrstka, S. Campuzano, M. Bartosik
A long non-coding RNA biomarker, PCA3, demonstrated excellent specificity for prostate cancer, and thus has become an interesting alternative to PSA monitoring. Its detection utilizes mostly reverse transcription PCR with optical detection, making the protocol longer and more expensive. To avoid PCR, we have developed an electrochemical assay coupled with LAMP, an isothermal amplification technique showing high sensitivities at constant temperatures and shorter reaction times. This is to our knowledge a first such attempt to apply electrochemistry to determine two RNA biomarkers directly in urine samples of prostate cancer patients in a minimally invasive diagnostics format.
Open Access – Carbon nanodot-based electrogenerated chemiluminescence biosensor for miRNA-21 detection
L. Gutiérrez-Gálvez, T. García-Mendiola, C. Gutiérrez-Sánchez, T. Guerrero-Esteban, C. García-Diego, I. Buendía, M. Laura García-Bermejo, F. Pariente and E. Lorenzo
A simple carbon nanodot-based electrogenerated chemiluminescence biosensor is described for sensitive and selective detection of microRNA-21 (miRNA-21), a biomarker of several pathologies including cardiovascular diseases (CVDs). The method does not require complex labeling steps, and has a rapid response. It was successfully used to detect miRNA-21 directly in serum samples from heart failure patients without previous RNA extraction neither amplification process.
A. Montero-Calle, Itziar Aranguren-Abeigon, M. Garranzo-Asensio, C. Poves, M.J. Fernández-Aceñero, J. Martínez-Useros, R. Sanz, J. Dziaková, J. Rodriguez-Cobos, G. Solís-Fernández, E. Povedano, M. Gamella, R.M. Torrente-Rodríguez, M. Alonso-Navarro, V. de los Ríos, J. Ignacio Casal, G. Domínguez, A. Guzman-Aranguez, A. Peláez-García, J.M. Pingarrón, S. Campuzano, R. Barderas
Autoantibodies against specific CRC autoantigens (tumor-associated antigens TAAs) in the sera of patients of colorectal cancer have been widely demonstrated to aid in early diagnosis. Thus, we herein aim to identify autoantigens target of autoantibodies specific to CRC that possess a significant ability to discriminate between CRC patients and healthy individuals by means of liquid biopsy. To that end, we examined the protein content of the exosomes released by five CRC cell lines and tissue samples from CRC patients by means of immunoprecipitation coupled with mass spectrometry analysis.
T. Guerrero-Esteban, C. Gutiérrez-Sánchez, T. García-Mendiola, M. Revenga-Parra, F. Pariente, E. Lorenzo
In this work, a promising disposable electrochemiluminescent immunosensor has been developed for simple, efficient detection of the HER2 protein, a breast cancer biomarker. The reliability of this biosensor was confirmed by analyzing HER2 in the presence of another tumor biomarker (CEA), as well as various proteins and sugars. In addition, this proposed strategy presented good stability and applicability in the analysis of human serum samples, showing great potential for applications in the early diagnosis of breast cancer.
Open Access – Electrochemical Immunosensing of ST2: A Checkpoint Target in Cancer Diseases
R. M. Torrente-Rodríguez, C. Muñoz-San Martín, M. Gamella, M. Pedrero, N. Martínez-Bosch, P. Navarro, P. García de Frutos, J. M. Pingarrón, S. Campuzano
A magnetic beads (MB)-involved amperometric immunosensor for the determination of ST2, a member of the IL 1 receptor family, es reported in this work. The method utilizar a sandwich immunoassay and disposable scree-printed carbon electrodes (SPCEs). The good correlation of the obtained results with those provided by an ELISA kit performed using the same immoreagentes demonstrates the potential of the developed strategy for early diagnosis and/or prognosis of the fatal PDAC disease.
B. Arévalo, A. ben Hassine, A. Valverde, V. Serafín, A. Montero-Calle, N. Raouafi, J. Camps, M. Arenas, R. Barderas, P. Yáñez-Sedeño, S. Campuzano, J.M. Pingarrón
Matrix metalloproteinase 9 (MMP-9) is a zinc-dependent endopeptidase that promotes angiogenesis, tumor growth, metastasis and cell invasion through the degradation of extracellular matrix. This work reports a magnetic microbeads (MBs)-based sandwich immunoassay for the amperometric determination of MMP-9 at screen-printed carbon electrodes (SPCEs). The results obtained show that the disposable and affordable immunoplatform developed is able not only to discriminate BC patients from healthy individuals but also to do it for the worst outcome triple negative (TNBC) subtype.
E. Povedano, M. Gamella, R. M. Torrente-Rodríguez, A. Montero-Calle, M. Pedrero, G. Solís-Fernández, F. Navarro-Villoslada, R. Barderas, S. Campuzano, J. M. Pingarrón
This work describes the preparation of an immunoplatform for the sensitive and selective determination of N6_methyladenosine (m6A). The simple and fast protocol involves for the first time the use of micromagnetic immunoconjugates to establish a direct competitive assay between the m6A target and a biotinylated RNA oligomer bearing a single m6A enzymatically labelled with a commercial conjugate of streptavidin-peroxidase (Strep-HRP) as tracer.
Open Access – Revisiting electrochemical biosensing in the 21st century society for inflammatory cytokines involved in autoimmune, neurodegenerative, cardiac, viral and cancer diseases – Review
S. Campuzano, P. Yáñez-Sedeño, J.M. Pingarrón
This review article puts forward electrochemical biosensing methods reported in the last five years for the determination of cytokines, summarizes recent developments and trends through a comprehensive discussion of selected strategies, and highlights the challenges to solve in this field. Considering the key role demonstrated in the last years by different materials (with nano or micrometric size and with or without magnetic properties), in the design of analytical performance-enhanced electrochemical biosensing strategies, special attention is paid to the methods exploiting these approaches.
Open Access – Breast cancer biomarker detection through the photoluminescence of epitaxial monolayer MoS2 flakes
S. Catalán-Gómez, M. Briones, S. Cortijo-Campos, T. García-Mendiola, A. de Andrés, S. Garg, P. Kung, E. Lorenzo, J. L. Pau, A. Redondo-Cubero
In this work we report on the characterization and biological functionalization of 2D MoS2 flakes, epitaxially grown on sapphire, to develop an optical biosensor for the breast cancer biomarker miRNA21. The results demonstrate the benefits of optical biosensors based on MoS2 monolayer for future commercial devices.
Influence of carbon nanodots on DNA-Thionine interaction. Application to breast cancer diagnosis
T. Garcia-Mendiola, S. Requena-Sanz, E. Martínez-Periñán, I. Bravo, F. Pariente, E. Lorenzo
In this work we present for the first time the effect of carbon nanodots on the interaction of DNA layers deposited on gold electrodes and Thionine. The acquired knowledge is applied to develop a new carbon nanodots based disposable DNA biosensor for the selective mutation detection in BRCA1 gene directly in clinical samples, using Thionine as redox indicator.
M. Zouari, S. Campuzano, J. M. Pingarrón, N. Raouafi
The preparation of an integrated biosensor for the easy, fast, and sensitive determination of miRNAs is described based on a direct hybridization format and a label-free voltammetric detection. The bioplatform allowed the determination of the target miRNA in a small amount of total RNA extracted from breast cancer (BC) cells or directly in serum samples collected from BC patients without the need for prior extraction, purification, amplification, or reverse transcription of the genetic material and with no matrix effect.
A. Valverde, V. Serafín, J. Garoz, A. Montero-Calle, A. González-Cort‚ s, M. Arenas, J. Camps, R. Barderas, P. Yañez-Sedeño, S. Campuzano, J.M. Pingarrón
This paper describes a dual immunosensor using neutravidin-functionalized magnetic microbeads (Neu-MBs) and dual screen-printed carbon electrodes (SPdCEs) for the simultaneous amperometric determination of two emerging biomarkers related to breast cancer (BC) and metastasis: Receptor Activator of Nuclear Factor-kB Ligand (RANKL) and Tumor Necrosis Factor alpha (TNF). The obtained results show the potential of this immunoplatform to improve the reliability of BC diagnosis using fast and cost-effective procedures.
Electrochemical biosensing to move forward in cancer epigenetics and metastasis: A review – Review
S. Campuzano, R. Barderas, M. Pedrero, P. Yáñez-Sedeño, J.M. Pingarrón
This review article critically discusses the main characteristics, opportunities and versatility exhibited by electrochemical biosensing, through highlighting representative examples published during the last two years, for the reliable determination of these emerging biomarkers, with great diagnostic, predictive and prognostic potencial.
A. Valverde, A. ben Hassine, V. Serafní, C. Muñoz-San Martín, M. Pedrero, M. Garranzo-Asensio, M. Gamella, N. Raouafi, R. Barderas, P. Yañez-Sedeño, S. Campuzano, J.M. Pingarrón
This paper reports the development of a dual immunosensor using magnetic microcarriers (MBs) and amperometric transduction at dual screen-printed carbon electrodes (SPdCEs) for the simultaneous determination of two biomarkers: interleukin-13 receptor alpha2 and E-cadherin (E-CDH), with both extracellular and soluble fraction; oncogenic and tumor suppressor markers, respectively, of great relevance in metastatic processes. The obtained results reveal great potential of this configuration to improve the reliability in diagnosing metastatic CRC.
M. Zouari, S. Campuzano, J.M. Pingarrón, N. Raouafi
A disposable enzyme-free biosensing platform for the sensitive and selective voltammetric determination of miRNAs is reported. The biosensor was able to determine accurately the target miRNA directly in scarcely diluted serum from breast cancer (BC) patients with no need for a previous total RNA extraction and in a very small amount of RNAt extracted from breast adenocarcinoma cells without the need for amplification or reverse transcription to complementary DNA.
Functionalization of a Few-Layer Antimonene with Oligonucleotides for DNA Sensing
T. García-Mendiola, C. Gutiérrez-Sánchez, C. Gibaja, I. Torres, C. Busó-Rogero, F. Pariente, J. Solera, Z. Razavifar, J.J. Palacios, F. Zamora, E. Lorenzo
Antimonene, a novel group 15 two-dimensional material, is functionalized with an oligonucleotide as a first step to DNA sensor development. The potential utility of this antimonene-based sensing device is assessed using, as a case of study, a sequence from the BRCA1 gene as the target DNA. The selectivity of the device allows not only recognition of a specific DNA sequence but also detection of a mutation in this gene associated with breast cancer, directly in clinical samples.
Magnetic beads-based electrochemical immunosensing of HIF-1α, a biomarker of tumoral hypoxia
C. Muñoz-San Martín, M. Gamella, M. Pedrero, A. Montero-Calle, R. Barderas, S. Campuzano, J.M. Pingarrón
This work reports the first magnetic beads (MBs)-based electrochemical immunoassay for the determination of Hypoxia-inducible factor-1 alpha, a transcription factor involver in tumor growth and metastasis by regulating genes involved in response to hypoxia. The design involves a sandwich immunoassay using MBs and amperometric detection at disposable screen-printed carbon electrodes (SPCEs).
A. Valverde, V. Serafin, A. Montero-Calle, A. González-Cortés‚ R. Barderas, P. Yañez-Sedeño, S. Campuzano, J.M. Pingarrón
This work reports the development of the first integrated electrochemical immunosensor for the determination of the ligand receptor activator nuclear factor-kB (RANKL), a biomarker that plays an important role in the regulation of bone resorption process and, as reported recently, in oncology by modulating the immune system response facilitating the tumor growth and metastatic process during cancer development. The developed immunosensor was applied to the determination of RANKL in human serum of patients diagnosed with colorectal cancer (CRC) or rheumatoid arthritis (RA).
M. Garranzo-Asensio, A. Guzmán, E. Povedano, V. Ruiz, C. Poves, M.J. Fernandez, A. Montero-Calle, G. Solís-Fernández, S. Fernandez, J. Campos, M. Arenas, N. Cabré, J. Joven, N. Rodriguez, G. Dominguez, P. Yáñez, J.M. Pingarrón, S. Campuzano, R. Barderas
The humoral immune response in cancer patients can be used for early detection of the disease. Autoantibodies raised against tumor-associated antigens (TAAs) are promising clinical biomarkers for reliable cancer diagnosis, prognosis, and therapy monitoring. In this study, an electrochemical disposable multiplexed immunosensing platform able to integrate difficult – and easy-to-express colorectal cancer (CRC) TAAs is reported for the sensitive determination of eight CRC-specific autoantibodies.
Nano/micromotors for diagnosis and therapy of cancer and infectious diseases – Review
K. Yuan, Z. Jiang, B. Jurado-Sánchez, A. Escarpa
This review highlights current progress and prospects in the use of nano and micromotors for diagnosis and treatment of infectious diseases and cancer. Motion-based sensing and fluorescence switching detection strategies along with therapeutic approaches based on direct cell capture; killing by direct contact or specific drug delivery to the affected site, will be comprehensively covered.
C. Muñoz San Martín, M. Pedrero, M. Gamella, A. Montero-Calle, R. Barderas, S. Campuzano, J.M. Pingarrón
Proteases are involved in cancer, taking part in immune (dis)regulation, malignant progression and tumour growth. Recently, it has been found that expression levels of one of the members of the serine protease family, trypsin, is upregulated in human cancer cells of several organs, being considered as a specific cancer biomarker. Considering the great attention that electrochemical peptide sensors have nowadays, in this work, we propose a novel electroanalytical strategy for the determination of this important biomolecule.
Advances in Electrochemical (Bio)Sensing Targeting Epigenetic Modifications of Nucleic Acids – Review
S. Campuzano, M. Pedrero, P. Yañez Sedeño, J.M. Pingarron
In this review article, a critical overview of recently reported advances in their development and use for the determination of epigenetic modification of nucleic acids, including the presence of methylated bases in DNA and RNA and the aberrant expression of microRNAs and IncRNAs, is provided. Special emphasis is made on methodologies successfully applied to real clinical samples, unmet challenges or key points towards future research.
Open Access – Opportunities, Challenges, and Prospects in Electrochemical Biosensing of Circulating Tumor DNA and Its Specific Features – Review
S. Campuzano, V. Serafín, M. Gamella, M. Pedrero, P. Yáñez-Sedeño, J. M. Pingarrón
Nowadays, analyzing circulating tumor DNA (ctDNA), a very small part of circulating free DNA (cfDNA) carried by blood, is considered to be an interesting alternative to conventional single-site tumor tissue biopsies. The determination of ctDNA and/or mapping its characteristic features, including tumor-specific mutations, chromosomal aberrations, microsatellite alterations, and epigenetic changes, are minimally invasive, powerful and credible biomarkers for early diagnosis, follow-up, prediction of therapy response/resistance, relapse monitoring, and tracking the rise of new mutant subclones, leading to improved cancer outcomes. This review provides an outline of advances published in the last five years in electrochemical biosensing of ctDNA and surrogate markers.
V. Serafín, A. Valverde, M. Garranzo-Asensio, R. Barderas, S. Campuzano, P. Yáñez-Sedeño & J. M. Pingarrón
This paper describes a dual electrochemical immunoassay for the simultaneous determination of IL-13Ralpha2 and CDH-17, two biomarkers of emerging relevance in metastatic processes. The simultaneous determination of these biomarkers was accomplished in lysates from breast and colorectal cancer cells with differents metastatic potential, and in paraffin-embedded tumot tissues extracts from patients diagnosed with colorectal cancer at different stages.
Open Access – Fluorescent C-NanoDots for rapid detection of BRCA1, CFTR and MRP3 gene mutations
T. García-Mendiola, C. Garcia Elosegui, I. Bravo, F. Pariente, A. Jacobo-Martin, C. Navio, Isabel Rodriguez, R. Wannemacher, E. Lorenzo
The authors report on a fluorometric method for the rapid detection of BRCA1, CFRT and MRP3 gene mutations. These are associated with breast cancer, cystic fibrosis and autoimmune hepatitios diseases, respectively. Carbon nanodots with blue fluorescence were synthesized and characterized, and their interactions with DNA were invetigated.