Sandwich-Type Electrochemical Paper-Based Immunosensor for Claudin 7 and CD81 Dual Determination on Extracellular Vesicles from Breast Cancer Patients

Ortega, Francisco G.; Regiart, Matías; Rodríguez–Martínez, A.; Pérez, Diego; Serrano, María José; Lorente, José A.; Tortella, Gonzalo; Rubilar, Olga; Sapag, Karim; Bertotti, Mauro; Fernández‐Baldo, Martín A.

doi:10.1021/acs.analchem.0c04180

Cited by 37 publications

(20 citation statements)

References 46 publications

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“…In the literature, several EVs associated proteins have been suggested as possible specific markers for detecting EVs released by cancer cells [ 10 ]. For example, EDIL3 [ 11 ] and Claudin7 [ 12 ] have been presented as possible makers of early breast cancer. Other studies, including smaller cohorts of subjects, also pointed out the possible use of EGRF [ 13 ], EpCAM [ 14 ], GPC-1 [ 15 ], CD82 [ 16 ], and Mucin-1 [ 17 ] present on the surface of cancer released EVs as possible diagnostic tools.…”

Section: Introductionmentioning

confidence: 99%

Fast quantification of extracellular vesicles levels in early breast cancer patients by Single Molecule Detection Array (SiMoA)

Morasso

Ricciardi

Sproviero

et al. 2021

Breast Cancer Res Treat

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Purpose Preliminary reports suggest that extracellular vesicles (EVs) might be a promising biomarker for breast cancer (BC). However, the quantification of plasmatic levels of EVs is a complex task. To overcome these limitations, we developed a new, fast, and easy to use assay for the quantification of EVs directly in plasma based on the use of Single-Molecule Array (SiMoA). Methods By using SiMoA to identify CD9+/CD63+ EVs, we analyzed plasma samples of 181 subjects (95 BC and 86 healthy controls, HC). A calibration curve, made of a serial dilution of lyophilized standards from human plasma, was used in each run to ensure the obtainment of quantitative results from the assay. In a subgroup of patients, EVs concentrations were estimated in plasma before and after 30 days from cancer surgery. Additional information on the size of EVs were also acquired using a Nanosight system to obtain a clearer understanding of the mechanism underlying the releases of EVs associated with the presence of cancer. Results The measured levels of EVs resulted significantly higher in BC patients (median values 1179.1 ng/µl vs 613.0 ng/µl, p < 0.0001). ROC curve was used to define the optimal cut-off level of the test at 1034.5 ng/µl with an AUC of 0.75 [95% CI 0.68–0.82]. EVs plasmatic concentrations significantly decreased after cancer surgery compared to baseline values (p = 0.014). No correlation was found between EVs concentration and clinical features of BC. Conclusion SiMoA assay allows plasmatic EVs levels detection directly without any prior processing. EVs levels are significantly higher in BC patients and significantly decreases after cancer surgery.

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Section: Introductionmentioning

confidence: 99%

Fast quantification of extracellular vesicles levels in early breast cancer patients by Single Molecule Detection Array (SiMoA)

Morasso

Ricciardi

Sproviero

et al. 2021

Breast Cancer Res Treat

View full text Add to dashboard Cite

show abstract

“…In addition, since the anti-CD63 has been widely used for the specific capture of sEVs, the specificity of P-ELISA is expected to be satisfactory. In addition, compared to other paper-based devices for EV detection (Table S4), our colorimetric instrument-free method possesses comparable sensitivity to other instrument-dependent methods (such as fluorescent, electrochemistry, , etc. ), ensuring its application for a rapid home test.…”

Section: Resultsmentioning

confidence: 93%

Highly Efficient Isolation and Sensitive Detection of Small Extracellular Vesicles Using a Paper-Based Device

et al. 2022

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Small extracellular vesicles (sEVs) play important roles in mediating intercellular communication and regulating biological processes. Facile sEV isolation is the essential and preliminary issue for their function investigation and downstream biomedical applications, while the traditional methods are challenged by tedious procedures, low purity, low yield, and potential damage. In this work, we developed an sEV isolation paper-based device (sEV-IsoPD) based on a three-dimensional (3D) paper chip, which is composed of a porous membrane for size exclusion and a metal–organic framework (MOF)/antibody-modified paper for immunoaffinity capture. In combination with a peristaltic pump-driven flow system, the sEV-IsoPD can efficiently isolate EV from cell culture medium and serum. Compared with the ultracentrifugation method, sEV-IsoPD exhibited a 5.1 times higher yield (1.7 × 109 mL–1), 1.6 times higher purity (1.6 × 1011 mg–1), and 7.5 times higher recovery (77.3%) with only 8.3% of the time (30 min) and 1.0% of the instrument cost ($710). Moreover, sEV concentration can be visually detected in a quantitative manner with this paper-based device with a linear range from 3.0 × 106 to 3.0 × 1010 mL–1 and a detection limit of 2.2 × 106 mL–1. The sEV-IsoPD provides an efficient and practical approach for the rapid isolation and visible detection of sEVs, which are promising for the preparation of sEVs and diagnosis of disease.

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“…Both claudin-7 and claudin-7/CD81 ratios were more accurate as diagnostic markers than the classic serum markers carcinoembryonic antigen (AUC, 0.5217 ± 0.068 SD) and mucin-1 (AUC, 0.7683 ± 0.0056 SD). Furthermore, claudin-7 and claudin-7/CD81 levels in EVs showed no significant association with ER, PR, and HER2 status in patients with BC (33), suggesting that claudin-7 might be a universal marker for the early diagnosis of BC. In addition, the presence of fibronectin on EVs derived from BC cells was revealed through proteomics.…”

Section: Candidate Ev Proteins For Early Bc Diagnosismentioning

confidence: 90%

Protein biomarkers in breast cancer-derived extracellular vesicles for use in liquid biopsies

Lai

Fan

et al. 2021

American Journal of Physiology-Cell Physiology

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Breast cancer is the most common malignant disease in women worldwide. Early diagnosis and treatment can greatly improve the management of breast cancer. Liquid biopsies are becoming convenient detection methods for diagnosing and monitoring breast cancer due to their non-invasiveness and ability to provide real-time feedback. A range of liquid biopsy markers, including circulating tumor proteins, circulating tumor cells, and circulating tumor nucleic acids, have been implemented for breast cancer diagnosis and prognosis, with each having its own advantages and limitations. Circulating extracellular vesicles are messengers of intercellular communication that are packed with information from mother cells and are found in a wide variety of bodily fluids; thus, they are emerging as ideal candidates for liquid biopsy biomarkers. In this review, we summarize extracellular vesicle protein markers that can be potentially used for the early diagnosis and prognosis of breast cancer or determining its specific subtypes.

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Sandwich-Type Electrochemical Paper-Based Immunosensor for Claudin 7 and CD81 Dual Determination on Extracellular Vesicles from Breast Cancer Patients

Cited by 37 publications

References 46 publications

Fast quantification of extracellular vesicles levels in early breast cancer patients by Single Molecule Detection Array (SiMoA)

Fast quantification of extracellular vesicles levels in early breast cancer patients by Single Molecule Detection Array (SiMoA)

Highly Efficient Isolation and Sensitive Detection of Small Extracellular Vesicles Using a Paper-Based Device

Protein biomarkers in breast cancer-derived extracellular vesicles for use in liquid biopsies

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