共查询到20条相似文献,搜索用时 31 毫秒
1.
Cristina Muoz‐SanMartín María Pedrero F. Javier ManueldeVillena María Garranzo‐Asensio Nuria Rodríguez Gemma Domínguez Rodrigo Barderas Susana Campuzano Jos M. Pingarrn 《Electroanalysis》2019,31(2):309-317
This paper describes the results obtained in the development of the first electrochemical immunosensor described to date for the detection of E‐cadherin (E‐cad) protein, a relevant biomarker of prognosis and metastasis in cancer, based on the use of magnetic microcarriers (MBs) and amperometric transduction at screen‐printed carbon electrodes (SPCEs). Thus, the determination of E‐cad protein involved the use of two specific antibodies against this protein (one of them labelled with HRP) in a sandwich configuration onto HOOC‐MBs. The magnetic bioconjugates were captured onto SPCEs and the amperometric transduction was performed using the H2O2/hydroquinone (HQ) system. Under optimal conditions, this bioplatform demonstrated a wide linear concentration range (0.50–25 ng mL?1) and a detection limit as low as 0.16 ng mL?1, well below the optimal cut‐off level for the E‐cad protein (defined as 10,000 ng mL?1 for soluble E‐cad levels in serum). The developed sensor also showed a good reproducibility among measurements with seven different sensors constructed in the same manner (RSD, 5.4 %), stability for more than 15 days and good specificity towards other proteins commonly found on biological samples. The applicability of this simple handling bioplatform for the direct determination of this protein in cell lysates with different metastatic potential and extracts from paraffined‐embedded human colorectal cancer tissues of different grade were also demonstrated. 相似文献
2.
Ludmila Jirakova Roman Hrstka Susana Campuzano Jose M. Pingarrn Martin Bartosik 《Electroanalysis》2019,31(2):293-302
There is an urgent need for development of rapid and inexpensive techniques for detection of microRNAs (miRNAs), which are potential biomarkers of various types of cancer. In this paper, we describe a multiplexed electrochemical platform for determination of three cancer‐relevant miRNAs: miR‐21, let‐7a and miR‐31. The strategy combines the use of magnetic beads (MBs) modified with a commercial antibody for the efficient capture of the heteroduplexes formed by hybridization of the target miRNA with DNA probe. Free non‐hybridized region of the DNA probe was thereafter hybridized with two biotin‐labeled auxiliary DNA probes in a process of hybridization chain reaction (HCR), resulting in a long hybrid bearing a large number of biotin molecules. Labeling of these multiple biotin units with streptavidin‐peroxidase conjugates allowed an amplification of the amperometric signal measured after capturing the modified MBs at a screen‐printed carbon electrode array of eight electrodes. The combined strategy demonstrated in a similar assay time significantly higher sensitivity than those previously described using modified MBs with the same capture antibody (without amplification by HCR) or a HCR strategy implemented on the surface of MBs, respectively. The methodology exhibits a good selectivity for discriminating single mismatches and was applied to the determination of the three target miRNAs in total RNA (RNAt) extracted from various cancer cell lines and from cervical precancerous lesions. 相似文献
3.
Beatriz Arévalo Verónica Serafín Susana Campuzano Paloma Yáñez-Sedeño José M. Pingarrón 《Electroanalysis》2021,33(9):2096-2104
This paper reports a dual immunosensor for the simultaneous determination of two important fertility-related hormones: 17β-estradiol, E2, and follicle-stimulating hormone, FSH. The implemented method involves direct competitive (E2) or sandwich-type (FSH) immunoassays carried out on magnetic microparticles (MBs) and amperometric detection at screen-printed dual carbon electrodes (SPdCEs) involving the hydroquinone (HQ)/H2O2 system. The developed immune platform demonstrates LOD values of 6.88 pg mL−1 and 0.11 mIU mL−1 for E2 and FSH standards, respectively and usefulness for the determination in saliva samples collected from different volunteers, giving results in agreement with the conventional ELISA methodologies. 相似文献
4.
Multiplexed Determination of Amino‐Terminal Pro‐B‐Type Natriuretic Peptide and C‐Reactive Protein Cardiac Biomarkers in Human Serum at a Disposable Electrochemical Magnetoimmunosensor 下载免费PDF全文
Berta Esteban‐Fernández de Ávila Vanessa Escamilla‐Gómez Valentina Lanzone Susana Campuzano María Pedrero Dario Compagnone José M. Pingarrón 《Electroanalysis》2014,26(2):254-261
A rapid magnetoimmunosensor for the simultaneous determination of two cardiac biomarkers, amino‐terminal pro‐B‐type natriuretic peptide (NT‐proBNP) and C‐reactive protein (CRP), in human serum is described. Specific capture antibodies were covalently immobilized onto carboxylic acid‐modified magnetic beads. The quantification of NT‐proBNP and CRP was performed by using indirect competitive and sandwich configurations, respectively, and horseradish peroxidase‐labeled tracers. The use of dual screen‐printed carbon electrodes allowed the achievement of simultaneous independent amperometric readout for each cardiac biomarker. The developed methodology showed very low limits of detection (0.47 ng mL?1). An international standard for CRP serum spiked with NT‐proBNP was analyzed to evaluate the usefulness of the magnetoimmunosensor. 相似文献
5.
《Electrophoresis》2018,39(12):1452-1459
Harvesting rare circulating tumor cells (CTCs) from human blood is distinctly substantial to monitor tumor stage and evaluate therapeutic efficacy. As a proof‐of‐concept study, a microfluidic chip with twin‐layer herringbone grooves was developed to isolate and recover tumor cells with high efficiency based on the immunoreaction between cells and antibody‐conjugated microbeads (MBs) under local magnetic field. Functional MBs were initially localized onto the internal channel wall through the magnetic guidance. Then, infused tumor cells were deviated into the herringbone groove via passive microvortex and were further trapped through an irreversible interaction with MBs. Upon the removal of magnet, the captured cells and residual MBs were released from the channel and collected for further analysis in cell adhesion and proliferation in vitro. Capture efficiency of tumor cells reached up to ∼90% and limit of detection was down to 50 cells per mL based on this approach. Furthermore, recovery rate of tumor cells was as high as ∼94%, and potencies of cell attachment and proliferation was well maintained in retrieved cells. Hence, the present technique has a great potential for the isolation, quantitation and recovery of CTCs for cancer theranostic guidance and biomolecular analysis. 相似文献
6.
Direct Determination of miR‐21 in Total RNA Extracted from Breast Cancer Samples Using Magnetosensing Platforms and the p19 Viral Protein as Detector Bioreceptor 下载免费PDF全文
S. Campuzano E. López‐Hernández R. Granados J. M. Sánchez‐Puelles J. M. Pingarrón 《Electroanalysis》2014,26(10):2080-2087
A novel magnetobiosensing approach for the rapid, sensitive and selective miR‐21 detection is reported involving the use of a specific RNA probe (antimiR‐21), streptavidin‐magnetic beads (Strep‐MBs), the siRNA Binding Protein p19 as detector bioreceptor, and amperometric detection with the H2O2/hydroquinone (HQ) system at disposable screen‐printed carbon electrodes. The magnetosensor exhibited a dynamic range from 1.4 to 10 nM and a detection limit of 4.2 fmol of synthetic target miR‐21 without any amplification step in 75 min. The usefulness of the approach was evaluated by analyzing total RNA (RNAt) extracted from metastatic breast cancer cell lines, human tissues and breast cytologies. 相似文献
7.
Irene Ojeda María Moreno‐Guzmán Araceli González‐Cortés Paloma Yáñez‐Sedeño José M. Pingarrón 《Electroanalysis》2013,25(9):2166-2174
Electrochemical immunosensors for ceruloplasmin (Cp) are reported for the first time. Two configurations involving magnetic beads (MBs) functionalized with Protein A or Streptavidin for immobilization of Cp antibodies were compared, using competitive immunoassay with synthesized alkaline phosphatase‐Cp conjugate. Upon capturing MBs‐immunoconjugates onto screen‐printed carbon electrodes, quantification of Cp was accomplished by DPV measurement of 1‐naphthol generated after 1‐naphthylphosphate addition. Linear ranges of calibration curves and detection limits were 0.1–1000 µg/mL and 0.040 µg/mL (Protein A‐MBs), and 0.025–20 µg/mL and 0.018 µg/mL (Strept‐MBs). Good results were obtained in the determination of Cp in spiked human serum samples. 相似文献
8.
Sensitive and selective magnetoimmunosensing platform for determination of the food allergen Ara h 1
V. Ruiz-Valdepeñas Montiel S. Campuzano A. Pellicanò R.M. Torrente-Rodríguez A.J. Reviejo M.S. Cosio J.M. Pingarrón 《Analytica chimica acta》2015
A highly sensitive disposable amperometric immunosensor based on the use of magnetic beads (MBs) is described for determination of Ara h 1, the major peanut allergen, in only 2 h. The approach uses a sandwich configuration involving selective capture and biotinylated detector antibodies and carboxylic acid-modified MBs (HOOC-MBs). The MBs bearing the immunoconjugates are captured by a magnet placed under the surface of a disposable screen-printed carbon electrode (SPCE) and the affinity reactions are monitored amperometrically at −0.20 V (vs a Ag pseudo-reference electrode) in the presence of hydroquinone (HQ) as electron transfer mediator and upon addition of H2O2 as the enzyme substrate. The developed immunosensor exhibits a wide range of linearity between 20.8 and 1000.0 ng mL−1 Ara h 1, a detection limit of 6.3 ng mL−1, a great selectivity, a good reproducibility with a RSD of 6.3% for six different immunosensors and a useful lifetime of 25 days. The usefulness of the immunosensor was demonstrated by determining Ara h 1 in different matrices (food extracts and saliva). The results correlated properly with those provided by a commercial ELISA method offering a reliable and promising analytical screening tool in the development of user-friendly devices for on-site determination of Ara h 1. 相似文献
9.
Targeted Ultrasound‐Assisted Cancer‐Selective Chemical Labeling and Subsequent Cancer Imaging using Click Chemistry 下载免费PDF全文
Hua Wang Marianne Gauthier Jamie R. Kelly Rita J. Miller Ming Xu Prof. Dr. William D. O'Brien Jr. Prof. Dr. Jianjun Cheng 《Angewandte Chemie (International ed. in English)》2016,55(18):5452-5456
Metabolic sugar labeling followed by the use of reagent‐free click chemistry is an established technique for in vitro cell targeting. However, selective metabolic labeling of the target tissues in vivo remains a challenge to overcome, which has prohibited the use of this technique for targeted in vivo applications. Herein, we report the use of targeted ultrasound pulses to induce the release of tetraacetyl N‐azidoacetylmannosamine (Ac4ManAz) from microbubbles (MBs) and its metabolic expression in the cancer area. Ac4ManAz‐loaded MBs showed great stability under physiological conditions, but rapidly collapsed in the presence of tumor‐localized ultrasound pulses. The released Ac4ManAz from MBs was able to label 4T1 tumor cells with azido groups and significantly improved the tumor accumulation of dibenzocyclooctyne (DBCO)‐Cy5 by subsequent click chemistry. We demonstrated for the first time that Ac4ManAz‐loaded MBs coupled with the use of targeted ultrasound could be a simple but powerful tool for in vivo cancer‐selective labeling and targeted cancer therapies. 相似文献
10.
Two different D‐dimer disposable amperometric immunosensing designs based on indirect competitive or sandwich formats and the use of carboxylic acid‐modified magnetic beads (COOH‐MBs) and screen‐printed carbon electrodes (SPCEs) have been developed and compared. In both approaches, the resulting modified MBs were magnetically captured on the surface of a SPCE which was used as the transducer for the electrochemical detection at ?0.20 V upon addition of H2O2, and hydroquinone (HQ). Both configurations exhibited linear ranges of clinical usefulness and detection limits quite below the clinical threshold (0.5 µg mL?1 D‐dimer). The sandwich configuration has been successfully tested with serum samples. 相似文献
11.
Magnetobiosensors Based on Viral Protein p19 for MicroRNA Determination in Cancer Cells and Tissues 下载免费PDF全文
Dr. Susana Campuzano Dr. Eva López‐Hernández Felipe Conzuelo Dr. Rosario Granados Dr. José M. Sánchez‐Puelles Prof. José M. Pingarrón 《Angewandte Chemie (International ed. in English)》2014,53(24):6168-6171
MicroRNAs (miRs) have emerged as important clinical biomarkers with both diagnostic and prognostic value for relevant diseases, such as cancer. MiRs pose unique challenges for detection and are currently detected by northern blotting, real‐time PCR, and microarray techniques. These expensive, complicated, and time‐consuming techniques are not feasible for on‐site miR determination. In this study, amperometric magnetobiosensors involving RNA‐binding viral protein p19 as a selective biorecognition element were developed for miR quantification. The p19‐based magnetosensors were able to detect 0.4 fmol of a synthetic target and endogenous miR‐21 (selected as a model for its role in a wide variety of cancers) in only 2 h in total RNA extracted from cancer cells and human breast‐tumor specimens without PCR amplification and sample preprocessing. These results open up formidable perspectives for the diagnosis and prognosis of human cancers and for drug‐discovery programs. 相似文献
12.
《Journal of mass spectrometry : JMS》2017,52(12):ii-ii
The identification of biomarkers would be of benefit for the diagnosis and treatment of colorectal cancer. DNA methylation in specific genomic regions, which had shown strongly association with disease genotypes, was an effective indicator to reveal the occurrence and development of cancers. To screen out methylation biomarkers for colorectal cancer (CRC), genomic DNA was isolated from colorectal cancerous and corresponding cancer‐adjacent tissues collected from 30 CRC patients and then bisulfite‐converted. The exon regions of 5 targeted genes (CNRIP1 , HIC1 , RUNX3 , p15 , and SFRP2 ) were amplified by using nested polymerase chain reaction with specific primers, and the amplicon was purified and hydrolyzed. The methylation levels of these specific regions were detected by liquid chromatography tandem mass spectrometry (LC‐MS/MS). The results showed that 5 targeted exon regions were successfully amplified and confirmed by sequencing. The methodological validations indicated that LC‐MS/MS was highly sensitive and accurate. The methylation levels of CNRIP1 and RUNX3 were remarkably high in CRC tissues with statistical difference when compared with corresponding cancer‐adjacent individuals, while that of HIC1 , p15 , and SFRP2 had no difference between 2 subjects. These findings supported CNRIP1 and RUNX3 as potential DNA methylation biomarkers for CRC diagnosis and treatment, and our LC‐MS/MS approach exhibited great advantages in the identification of regional DNA methylation biomarkers. 相似文献
13.
Electrochemical biosensors are highly compatible with modern advancements in magnetic nanomaterials. In particular, the versatile nature of magnetic nanomaterials as a universal platform for selective isolation of diverse forms of cancer biomarkers in body circulation, is highly synergistic with electrochemical biosensors for elevating biosensing performance to unprecedented levels. Such diverse circulating target biomolecules include cell surface proteins of circulating tumor cells and extracellular vesicles (EVs), as well as circulating tumor nucleic acids (i.e. ctDNA/ctRNA). This focussed review serves to discuss the latest work in the fields of magnetic nanomaterials and electrochemistry to tackle existing analysis challenges of diverse circulating biomarkers in cancer. 相似文献
14.
《Electrophoresis》2018,39(11):1375-1381
Methyl parathion (MP) is a highly toxic organophosphate and its exposure may lead to substantial adverse effects to human health. The existence of 4‐nitrophenol (4‐NP) in the form of free phenol, glucuronide (4‐NP‐G) or as a sulfate ester (4‐NP‐S) can be used as biomarkers to assess the duration and extent of MP exposure. In this work, a MC‐CE device incorporating post‐CE amperometric detection using multi‐walled carbon nanotubes (MWNTs) modified carbon fiber microelectrode (CFME) was fabricated and assessed for simultaneous determination of 4‐NP, 4‐NP‐G, and 4‐NP‐S in human urine. The detection sensitivity and stability was greatly enhanced by the modification of MWNTs. The capability of the MC‐CE device with dual MWNTs modified CFME for detecting impurity was assessed and reliability established by high recoveries from 95 to 97% for spiked MP biomarkers. The method developed is shown to provide a simple, sensitive, and reliable means for monitoring 4‐NP, 4‐NP‐G, and 4‐NP‐S in human urine. 相似文献
15.
Ling‐Ling Wu Zhi‐Ling Zhang Man Tang Dong‐Liang Zhu Xiao‐Juan Dong Jiao Hu Chu‐Bo Qi Hong‐Wu Tang Dai‐Wen Pang 《Angewandte Chemie (International ed. in English)》2020,59(28):11240-11244
Comprehensive phenotypic profiling of heterogeneous circulating tumor cells (CTCs) at single‐cell resolution has great importance for cancer management. Herein, a novel spectrally combined encoding (SCE) strategy was proposed for multiplex biomarker profiling of single CTCs using a multifunctional nanosphere‐mediated microfluidic platform. Different cellular biomarkers uniquely labeled by multifunctional nanosphere barcodes, possessing identical magnetic tags and distinct optical signatures, enabled isolation of heterogeneous CTCs with over 91.6 % efficiency and in situ SCE of phenotypes. By further trapping individual CTCs in ordered microstructures on chip, composite single‐cell spectral signatures were conveniently and efficiently obtained, allowing reliable spectral‐readout for multiplex biomarker profiling. This SCE strategy exhibited great potential in multiplex profiling of heterogeneous CTC phenotypes, offering new avenues for cancer study and precise medicine. 相似文献
16.
U. Eletxigerra J. Martinez-Perdiguero S. Merino R. Villalonga J.M. Pingarrón S. Campuzano 《Analytica chimica acta》2014
An amperometric immunoassay for the determination of tumor necrosis factor alpha (TNFα) protein biomarker in human serum based on the use of magnetic microbeads (MBs) and disposable screen-printed carbon electrodes (SPCEs) has been developed. The specifically modified microbeads were magnetically captured on the working electrode surface and the amperometric responses were measured at −0.20 V (vs. Ag pseudo-reference electrode), upon addition of hydroquinone (HQ) as electron transfer mediator and H2O2 as the enzyme substrate. After a thorough optimization of the assay, extremely low limits of detection were achieved: 2.0 pg mL−1 (36 fM) and 5.8 pg mL−1 (105 fM) for standard solutions and spiked human serum, respectively. The simplicity, robustness and this clinically interesting LOD proved the developed TNFα immunoassay as a good contender for real clinical application. 相似文献
17.
Electrochemical Biosensors for Cancer Biomarkers Detection: Recent Advances and Challenges 下载免费PDF全文
Biomarkers are described as characteristics that provide information about biological conditions whether normal or pathological. Detection of biomarkers at the earliest stage of the cancer is of utmost importance for clinical diagnosis. Electrochemical biosensors allow detecting the low levels of specific analytes in blood, urine or saliva and providing a sensitive approach for direct measurement for cancer biomarker detection. Moreover, the integration of electrochemical devices with nanomaterials, such as carbon nanotubes, gold and magnetic particles offer amplification and multiplexing capabilities for simultaneous measurements of cancer biomarkers very sensitively. This review summarizes the recent developments of electrochemical biosensors systems for the detection of cancer biomarkers with emphasis on voltammetric, amperometric and impedimetric biosensors. A special attention is paid to aptamers and miRNAs that are very promising for the ultra‐sensitive and specific cancer biomarker detection. 相似文献
18.
《Biomedical chromatography : BMC》2017,31(11)
Colorectal cancer (CRC) is one of the most common human malignancies and encompasses cancers of the colon and rectum. Although the gold‐standard colonoscopy screening method is effective in detecting CRC, this method is invasive and can result in severe complications for patients. The purpose of this study was to determine differences in metabolites between CRC and matched adjacent nontumor tissues from CRC patients, to identify potential biomarkers that may be informative and developed screening methods. Metabolomic analysis was performed on clinically localized CRC tissue and matched adjacent nontumor tissue from 20 CRC patients. Unsupervised analysis, supervised analysis, univariate analysis and pathway analysis were used to identify potential metabolic biomarkers of CRC. The levels of 25 metabolites in CRC tissues were significantly altered compared with the matched adjacent nontumor tissues. Four metabolites (lactic acid, alanine, phosphate and aspartic acid) demonstrated good area under the curve of receiver‐operator characteristic with acceptable sensitivities and specificities, indicating their potential as important biomarkers for CRC. Alterations of amino acid metabolism and enhanced glycolysis may be major factors in the development and progression of CRC. Lactic acid, alanine, phosphate, and aspartic acid could be effective diagnostic indicators for CRC. 相似文献
19.
Structurally Defined αMHC‐II Nanobody–Drug Conjugates: A Therapeutic and Imaging System for B‐Cell Lymphoma 下载免费PDF全文
Dr. Tao Fang Dr. Joao N. Duarte Jingjing Ling Zeyang Li Jonathan S. Guzman Prof. Dr. Hidde L. Ploegh 《Angewandte Chemie (International ed. in English)》2016,55(7):2416-2420
Antibody–drug conjugates (ADCs) of defined structure hold great promise for cancer therapies, but further advances are constrained by the complex structures of full‐sized antibodies. Camelid‐derived single‐domain antibody fragments (VHHs or nanobodies) offer a possible solution to this challenge by providing expedited target screening and validation through switching between imaging and therapeutic activities. We used a nanobody (VHH7) specific for murine MHC‐II and rendered “sortase‐ready” for the introduction of oligoglycine‐modified cytotoxic payloads or NIR fluorophores. The VHH7 conjugates outcompeted commercial monoclonal antibodies (mAbs) for internalization and exhibited high specificity and cytotoxicity against A20 murine B‐cell lymphoma. Non‐invasive NIR imaging with a VHH7–fluorophore conjugate showed rapid tumor targeting on both localized and metastatic lymphoma models. Subsequent treatment with the nanobody–drug conjugate efficiently controlled tumor growth and metastasis without obvious systemic toxicity. 相似文献
20.
Esteban-Fernández de Ávila B Pedrero M Campuzano S Escamilla-Gómez V Pingarrón JM 《Analytical and bioanalytical chemistry》2012,403(4):917-925
The preparation and characteristics of a disposable amperometric magnetoimmunosensor, based on the use of functionalized magnetic
beads (MBs) and gold screen-printed electrodes (Au/SPEs), for the specific detection and quantification of Staphylococcal
protein A (ProtA) and Staphylococcus aureus (S. aureus) is reported. An antiProtA antibody was immobilized onto ProtA-modified MBs, and a competitive immunoassay involving ProtA
antigen labelled with HRP was performed. The resulting modified MBs were captured by a magnetic field on the surface of tetrathiafulvalene-modified
Au/SPEs and the amperometric response obtained at −0.15 V vs the silver pseudo-reference electrode of the Au/SPEs after the
addition of H2O2 was used as transduction signal. The developed methodology showed very low limits of detection (1 cfu S. aureus/mL of raw milk samples), and a good selectivity against the most commonly involved foodborne pathogens originating from milk.
These features, together with a short analysis time (2 h), the simplicity, and easy automation and miniaturization of the
required instrumentation make the developed methodology a promising alternative in the development of devices for on-site
analysis. 相似文献