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1.
Biomed Pharmacother ; 176: 116826, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38838507

RESUMEN

BACKGROUND: Phosphatidylinositol-4-phosphate 5-kinase type 1 alpha (PIP5K1A) acts upstream of the Akt regulatory pathway and is abnormally expressed in many types of malignancies. However, the role and mechanism of PIP5K1A in colorectal cancer (CRC) have not yet been reported. In this study, we aimed to determine the association between PIP5K1A and progression of CRC and assess the efficacy and mechanism by which rupatadine targets PIP5K1A. METHODS: Firstly, expression and function of PIP5K1A in CRC were investigated by human colon cancer tissue chip analysis and cell proliferation assay. Next, rupatadine was screened by computational screening and cytotoxicity assay and interactions between PIP5K1A and rupatadine assessed by kinase activity detection assay and bio-layer interferometry analysis. Next, rupatadine's anti-tumor effect was evaluated by in vivo and in vitro pharmacodynamic assays. Finally, rupatadine's anti-tumor mechanism was explored by quantitative real-time reverse-transcription polymerase chain reaction, western blot, and immunofluorescence. RESULTS: We found that PIP5K1A exerts tumor-promoting effects as a proto-oncogene in CRC and aberrant PIP5K1A expression correlates with CRC malignancy. We also found that rupatadine down-regulates cyclin-dependent kinase 2 and cyclin D1 protein expression by inhibiting the PIP5K1A/Akt/GSK-3ß pathway, induces cell cycle arrest, and inhibits CRC cell proliferation in vitro and in vivo. CONCLUSIONS: PIP5K1A is a potential drug target for treating CRC. Rupatadine, which targets PIP5K1A, could serve as a new option for treating CRC, its therapeutic mechanism being related to regulation of the Akt/GSK-3ß signaling pathway.


Asunto(s)
Proliferación Celular , Neoplasias Colorrectales , Ciproheptadina , Fosfotransferasas (Aceptor de Grupo Alcohol) , Proteínas Proto-Oncogénicas c-akt , Transducción de Señal , Humanos , Proliferación Celular/efectos de los fármacos , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/patología , Neoplasias Colorrectales/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Animales , Transducción de Señal/efectos de los fármacos , Fosfotransferasas (Aceptor de Grupo Alcohol)/metabolismo , Ciproheptadina/farmacología , Ciproheptadina/análogos & derivados , Ratones Desnudos , Línea Celular Tumoral , Ratones Endogámicos BALB C , Masculino , Proto-Oncogenes Mas , Ensayos Antitumor por Modelo de Xenoinjerto , Ratones , Antineoplásicos/farmacología
2.
ACS Sens ; 9(6): 2897-2906, 2024 Jun 28.
Artículo en Inglés | MEDLINE | ID: mdl-38776471

RESUMEN

Ovarian cancer (OC) has the highest mortality rate among malignant tumors, primarily because it is difficult to diagnose early. Exosomes, a type of extracellular vesicle rich in parental information, have garnered significant attention in the field of cancer diagnosis and treatment. They play an important regulatory role in the occurrence, development, and metastasis of OC. Consequently, exosomes have emerged as noninvasive biomarkers for early cancer detection. Therefore, identifying cancer-derived exosomes may offer a novel biomarker for the early detection of OC. In this study, we developed a metal-organic frameworks assembled "double hook"-type aptamer electrochemical sensor, which enables accurate early diagnosis of OC. Under optimal experimental conditions, electrochemical impedance spectroscopy technology demonstrated a good linear relationship within the concentration range of 31-3.1 × 106 particles per microliter, with a detection limit as low as 12 particles per microliter. The universal exosome detection platform is constructed, and this platform can not only differentiate between high-grade serous ovarian cancer (HGSOC) patients and healthy individuals but also distinguish between HGSOC patients and nonhigh-grade serous OC (non-HGSOC). Consequently, it provides a novel strategy for the early diagnosis of OC and holds great significance in clinical differential diagnosis.


Asunto(s)
Detección Precoz del Cáncer , Neoplasias Ováricas , Femenino , Neoplasias Ováricas/diagnóstico , Humanos , Detección Precoz del Cáncer/métodos , Técnicas Electroquímicas/métodos , Técnicas Biosensibles/métodos , Aptámeros de Nucleótidos/química , Estructuras Metalorgánicas/química , Exosomas/química , Límite de Detección , Espectroscopía Dieléctrica/métodos , Biomarcadores de Tumor/análisis
3.
Anal Methods ; 16(19): 3039-3046, 2024 May 16.
Artículo en Inglés | MEDLINE | ID: mdl-38682261

RESUMEN

Beta-lactoglobulin (ß-Lg), a prominent milk protein, is a major contributor to milk allergies. The quantitative assessment of ß-Lg is a valuable method for assessing the allergenic potential of dairy products. In this study, a specific aptamer, ß-Lg-01, with an affinity constant (KD) of 28.6 nM for ß-Lg was screened through seven rounds of magnetic bead SELEX (MB-SELEX). A novel bio-layer interferometry (BLI)-based aptasensor was developed, which had a limit of detection (LOD) of 0.3 ng mL-1, a linear range of 1.5 ng mL-1-15 µg mL-1, and a recovery rate of 102-116% among the milk samples. This aptasensor provides a potential tool for the detection and risk assessment of ß-Lg within 10 min.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Lactoglobulinas , Leche , Técnica SELEX de Producción de Aptámeros , Lactoglobulinas/análisis , Lactoglobulinas/química , Leche/química , Técnicas Biosensibles/métodos , Animales , Aptámeros de Nucleótidos/química , Técnica SELEX de Producción de Aptámeros/métodos , Límite de Detección , Interferometría/métodos
4.
Molecules ; 29(8)2024 Apr 13.
Artículo en Inglés | MEDLINE | ID: mdl-38675595

RESUMEN

The COVID-19 pandemic over recent years has shown a great need for the rapid, low-cost, and on-site detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this study, an aptamer-based colloidal gold nanoparticle lateral flow test strip was well developed to realize the visual detection of wild-type SARS-CoV-2 spike proteins (SPs) and multiple variants. Under the optimal reaction conditions, a low detection limit of SARS-CoV-2 S proteins of 0.68 nM was acquired, and the actual detection recovery was 83.3% to 108.8% for real-world samples. This suggests a potential tool for the prompt detection of SARS-CoV-2 with good sensitivity and accuracy, and a new method for the development of alternative antibody test strips for the detection of other viral targets.


Asunto(s)
Aptámeros de Nucleótidos , COVID-19 , Oro , Nanopartículas del Metal , SARS-CoV-2 , Glicoproteína de la Espiga del Coronavirus , Humanos , Aptámeros de Nucleótidos/química , COVID-19/diagnóstico , COVID-19/virología , Oro/química , Límite de Detección , Nanopartículas del Metal/química , Tiras Reactivas , SARS-CoV-2/química , Glicoproteína de la Espiga del Coronavirus/química
5.
Biosens Bioelectron ; 257: 116313, 2024 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-38688229

RESUMEN

The emergence and rapid spread of Mpox (formerly monkeypox) have caused significant societal challenges. Adequate and appropriate diagnostics procedures are an urgent necessity. Herein, we discover a pair of aptamers through the systematic evolution of ligands by exponential enrichment (SELEX) that exhibit high affinity and bind to different sites towards the A29 protein of the Mpox virus. Subsequently, we propose a facile, sensitive, convenient CRISPR/Cas12a-mediated aptasensor for detecting the A29 antigen. The procedure employs the bivalent aptamers recognition, which induces the formation of a proximity switch probe and initiates subsequent cascade strand displacement reactions, then triggers CRISPR/Cas12a DNA trans-cleavage to achieve the sensitive detection of Mpox. Our method enables selective and ultrasensitive evaluation of the A29 protein within the range of 1 ng mL-1 to 1 µg mL-1, with a limit of detection (LOD) at 0.28 ng mL-1. Moreover, spiked A29 protein recovery exceeds 96.9%, while the detection activity remains above 91.9% after six months of storage at 4 °C. This aptasensor provides a novel avenue for exploring clinical diagnosis in cases involving Mpox as facilitating development in various analyte sensors.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Sistemas CRISPR-Cas , Límite de Detección , Técnica SELEX de Producción de Aptámeros , Técnicas Biosensibles/métodos , Aptámeros de Nucleótidos/química , Humanos , Antígenos Virales/análisis , Proteínas Asociadas a CRISPR/química , Proteínas Bacterianas , Endodesoxirribonucleasas
6.
Nano Lett ; 23(24): 11438-11446, 2023 Dec 27.
Artículo en Inglés | MEDLINE | ID: mdl-38051760

RESUMEN

Single-molecule antigen detection using nanopores offers a promising alternative for accurate virus testing to contain their transmission. However, the selective and efficient identification of small viral proteins directly in human biofluids remains a challenge. Here, we report a nanopore sensing strategy based on a customized DNA molecular probe that combines an aptamer and an antibody to enhance the single-molecule detection of mpox virus (MPXV) A29 protein, a small protein with an M.W. of ca. 14 kDa. The formation of the aptamer-target-antibody sandwich structures enables efficient identification of targets when translocating through the nanopore. This technique can accurately detect A29 protein with a limit of detection of ∼11 fM and can distinguish the MPXV A29 from vaccinia virus A27 protein (a difference of only four amino acids) and Varicella Zoster Virus (VZV) protein directly in biofluids. The simplicity, high selectivity, and sensitivity of this approach have the potential to contribute to the diagnosis of viruses in point-of-care settings.


Asunto(s)
Mpox , Nanoporos , Humanos , Proteínas/química , Nanotecnología/métodos , ADN/química , Anticuerpos , Oligonucleótidos
7.
Nat Commun ; 14(1): 7362, 2023 11 14.
Artículo en Inglés | MEDLINE | ID: mdl-37963924

RESUMEN

We report on single-molecule nanopore sensing combined with position-encoded DNA molecular probes, with chemistry tuned to simultaneously identify various antigen proteins and multiple RNA gene fragments of SARS-CoV-2 with high sensitivity and selectivity. We show that this sensing strategy can directly detect spike (S) and nucleocapsid (N) proteins in unprocessed human saliva. Moreover, our approach enables the identification of RNA fragments from patient samples using nasal/throat swabs, enabling the identification of critical mutations such as D614G, G446S, or Y144del among viral variants. In particular, it can detect and discriminate between SARS-CoV-2 lineages of wild-type B.1.1.7 (Alpha), B.1.617.2 (Delta), and B.1.1.539 (Omicron) within a single measurement without the need for nucleic acid sequencing. The sensing strategy of the molecular probes is easily adaptable to other viral targets and diseases and can be expanded depending on the application required.


Asunto(s)
Antígenos Virales , Nanoporos , Humanos , Antígenos Virales/genética , Sondas Moleculares , ARN , ARN Viral/genética
8.
Anal Methods ; 15(16): 2039-2043, 2023 04 27.
Artículo en Inglés | MEDLINE | ID: mdl-37066673

RESUMEN

In this study, we constructed and optimized a semi-automatic instrument to perform aptamer SELEX targeting multiple proteins simultaneously. Our work provides a simple SELEX platform characterized by real-time feedback, which is time efficient and can reduce human intervention. A number of aptamers were rapidly screened by this method. Moreover, the binding affinities of these aptamers were verified by various methods, including SPR and flow cytometry, which supports the applicability and reliability of our newly established aptamer SELEX system.


Asunto(s)
Aptámeros de Nucleótidos , Humanos , Aptámeros de Nucleótidos/química , Técnica SELEX de Producción de Aptámeros/métodos , Reproducibilidad de los Resultados , Citometría de Flujo
9.
Talanta ; 256: 124312, 2023 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-36738621

RESUMEN

The CRISPR/Cas systems have provided wide biosensing applications. Particularly, the aptamer-involved CRISPR/Cas sensor system powerfully expanded to non-nucleic-acid targets. However, tailoring the sequence of the aptamer to explore the relationship between affinity and the activation of CRISPR/Cas12a trans-cleavage activity has not been reported yet. Herein, we developed a series of new aptamers toward the spike protein 1(S1) of SARS-CoV-2. Surface plasmon resonance measurements showed that the affinity of these aptamers to S1 was at the nM level. Subsequently, a "SET" effect (Sequence Essential Trans-cleavage activity) is discovered for the activation of CRISPR/Cas12a trans-cleavage activity. That is, an aptamer, as the activator, sequence needs to be tailored to activate CRISPR/Cas12a efficiently. A balance should be reached between affinity and activation ability. On the one hand, high affinity ensures target recognition performance, and on the other hand, activation can achieve adequate amplification and output of recognition signals. The optimized sequence (with 27 nucleotides, for short 27-nt) not only recognizes the target with a high affinity and specificity but also can trigger the CRISPR/Cas12a trans-cleavage activity efficiently, showing an excellent detection performance in electrochemical biosensors. The detection limit for SARS-CoV-2 S1 can be low at 1.5 pg mL-1. The new CRISPR/Cas12a-derived aptasensor also displays a remarkable ability to detect Beta, Delta, and Omicron variants but is selective toward other kinds of proteins. Above all, it is robust for point-of-care testing (POCT) in complex biological fluids, such as saliva, urine, and serum, and provides a universal and scalable detecting platform. Our results provide new insights into aptamer development and a different strategy for COVID-19 antigen detection and biosensor development.


Asunto(s)
Técnicas Biosensibles , COVID-19 , Humanos , COVID-19/diagnóstico , Sistemas CRISPR-Cas , SARS-CoV-2/genética , Oligonucleótidos , Resonancia por Plasmón de Superficie
10.
Sens Actuators B Chem ; 380: 133387, 2023 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-36694572

RESUMEN

Antigen-detecting rapid diagnostic testing (Ag-RDT) has contributed to containing the spread of SARS-CoV-2 variants of concern (VOCs). In this study, we proposed a biomimetic clamp assay for impedimetric SARS-CoV-2 nucleocapsid protein (Np) detection. The DNA biomimetic clamp (DNA-BC) is formed by a pair of Np aptamers connected via a T20 spacer. The 5'- terminal of the DNA-BC is phosphate-modified and then anchored on the surface of the screen-printed gold electrode, which has been pre-coated with Au@UiO-66-NH2. The integrated DNA-material sensing biochip is fabricated through the strong Zr-O-P bonds to form a clamp-type impedimetric aptasensor. It is demonstrated that the aptasensor could achieve Np detection in one step within 11 min and shows pronounced sensitivity with a detection limit of 0.31 pg mL-1. Above all, the aptasensor displays great specificity and stability under physiological conditions as well as various water environments. It is a potentially promising strategy to exploit reliable Ag-RDT products to confront the ongoing epidemic.

11.
Crit Rev Anal Chem ; 53(7): 1433-1454, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-35085047

RESUMEN

In view of ever-increasing environmental pollution, there is an immediate requirement to promote cheap, multiplexed, sensitive and fast biosensing systems to monitor these pollutants or contaminants. Aptamers have shown numerous advantages in being used as molecular recognition elements in various biosensing devices. Graphene and graphene-based materials/nanohybrids combined with several detection methods exhibit great potential owing to their exceptional optical, electronic and physicochemical properties which can be employed extensively to monitor environmental contaminants. For environmental monitoring applications, aptamers have been successfully combined with graphene-based nanohybrids to produce a wide range of innovative methodologies. Aptamers are immobilized at the surface of graphene based nanohybrids via covalent and non-covalent strategies. This review highlights the design, working principle, recent developmental advances and applications of graphene based nanohybrid aptasensors (GNH-Apts) (since January 2014 to September 2021) with a special emphasis on two major signal-transduction methods, i.e., optical and electrochemical for the monitoring of pesticides, heavy metals, bacteria, antibiotics, and organic compounds from different environmental samples (e.g., water, soil and related). Lastly, the challenges confronted by scientists and the possible future outlook have also been addressed. It is expected that high-performance graphene-based nanohybrid aptasensors would find broad applications in the field of environmental monitoring.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Grafito , Plaguicidas , Grafito/química , Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , Monitoreo del Ambiente
12.
Biosens Bioelectron ; 215: 114509, 2022 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-35940005

RESUMEN

Despite their high analytical performance, conventional analytical biosensor devices are usually difficult to handle, time-consuming, bulky and expensive. As a result, their applications remain restricted to resource-limited environments. In particular, the transportation of conventional analytical equipment is challenging for the proper in-situ point of need (PON)/point of care (POC) detection of biomolecules. In this context, smartphones, the most widely utilized cutting-edge mobile gadgets, continue to be a favored option because of their ease of portability and revolutionary sensing capabilities. On the other hand, aptamers are molecular recognition units consisting of nucleic acids with highly sensitive and selective recognition capabilities towards their respective targets. The coupling of smartphones with aptamers have led the development of advanced, user-friendly, portable, and cost-effective in-situ PON/POC biosensors for the detection of biomolecules. Such sensors are well-suited for a variety of applications, including food safety, environmental monitoring, and disease diagnostics. Herein, for the first time, achievements made between 2017 and 2022 in the concept and design of the smartphone-enabled aptasensors for biosensing applications are reviewed. The review covers different fabrication strategies and the discussion of several operating systems, underlying programs, and related software. At the end, the important merits, challenges, and future prospects of smart phone-driven aptasensors are presented. This report intends to assist scientists and engineers in comprehending the fabrication of smartphone-based aptasensors and their underlying sensing principles, as well as stimulating the future developments in the direction of affordable, portable, simple, and readily available sensing devices.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Sistemas de Atención de Punto , Teléfono Inteligente
13.
Biosens Bioelectron ; 213: 114436, 2022 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-35716641

RESUMEN

The emergence of the COVID-19 epidemic has affected the lives of hundreds of millions of people globally. There is no doubt that the development of fast and sensitive detection methods is crucial while the worldwide effective vaccination programs are miles away from actualization. In this study, we have reported an electrochemical N protein aptamer sensor with complementary oligonucleotide as probe for the specific detection of COVID-19. The electrochemical aptasensor was prepared by fixing the double-stranded DNA hybrid obtained by the hybridization of N protein aptamer and its Fc-labeled complementary strand on the surface of a gold electrode. After incubation with the target, the aptamer dissociated from the labeled complementary DNA oligonucleotide hybrid to preferentially bind with N protein in the solution. The concentration of N protein was measured by detecting the changes in electrochemical current signals induced by the conformational transformation of the complementary DNA oligonucleotide left on the electrode surface. The sensor had a linear relationship between the logarithm of the N protein concentration from 10 fM to 100 nM (ΔIp = 0.098 log CN protein/fM - 0.08433, R2 = 0.99), and the detection limitation was 1 fM (S/N = 3). The electrochemical aptamer sensor was applied to test the spiked concentrations of throat swabs and blood samples from three volunteers, and the obtained results proved that the sensor has great potentials for the early detection of COVID-19 in patients.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , COVID-19 , Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , COVID-19/diagnóstico , ADN Complementario , Técnicas Electroquímicas/métodos , Electrodos , Oro/química , Humanos , Límite de Detección , Unión Proteica
14.
Talanta ; 242: 123302, 2022 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-35180537

RESUMEN

In this paper, we report the development of a new strategy termed integrated digital PCR-fluorescence activated sorting based SELEX (IFS-SELEX) that enables rapid screening of aptamers against fluorescent targets. Initially, this strategy employs an integrated digital PCR system to amplify each sequence of a preliminarily enriched library, which is obtained by a traditional SELEX method, on the surface of polystyrene beads. Then, the as-prepared beads are incubated with the fluorescent target and then subjected to fluorescence-activated sorting. Since only those sequences with high binding affinity for the target are collected and sequenced, unnecessary analysis of ineligible sequences is avoided by this method, and the aptamer selection process is thereby greatly streamlined. As a proof-of-concept, we applied this strategy for the screening of aptamers against two fluorescent targets, i.e., ciprofloxacin (CFX) and thioflavin T (ThT), and successfully obtained corresponding sequences with low dissociation constants. The binding affinities of aptamers for ThT were well associated with the sorting regions defined in the fluorescence channel of the flow cytometry process. The experimental results demonstrated that the as-designed IFS-SELEX method can serve as a universal platform for rapid, facile, and efficient aptamer selection against various fluorescent targets.


Asunto(s)
Aptámeros de Nucleótidos , Técnica SELEX de Producción de Aptámeros , Aptámeros de Nucleótidos/genética , Aptámeros de Nucleótidos/metabolismo , Colorantes , Citometría de Flujo/métodos , Reacción en Cadena de la Polimerasa , Técnica SELEX de Producción de Aptámeros/métodos
15.
Biosens Bioelectron ; 200: 113922, 2022 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-34990959

RESUMEN

Fast, affordable, portable, and sensitive technology to detect COVID-19 is critical to address the current outbreak. Here, we present a CRISPR/Cas12a-derived electrochemical aptasensor for cost-effective, fast, and ultrasensitive COVID-19 nucleocapsid protein (Np) detection. First, an electrochemical sensing interface was fabricated by immobilizing methylene blue labeled poly adenines DNA sequence (polyA-MB electrochemical reporter) on a gold electrode surface. Second, an arched probe was prepared via hybridization of Np aptamer and an activator strand. In the presence of COVID-19 Np, the activator strand could be released from the arched probe due to the specific interaction between the target and the aptamer, which then activated the trans-cleavage activity of the CRISPR/Cas12a system. Subsequently, the polyA-MB reporters were cleaved from the electrode surface, decreasing the current of differential pulse voltammetry (DPV) at a potential of -0.27 V(vs. Ag/AgCl). The CRISPR/Cas12a-derived electrochemical aptasensor shows a highly efficient performance for COVID-19 Np detection in 50 pg mL-1 to 100 ng mL-1 with a limit of detection (LOD) low to 16.5 pg mL-1. Notably, the whole process of one test can be completed within 30 min. Simultaneously, the aptasensor displays a high selectivity to other proteins. The further measurements demonstrate that the aptasensor is robust in a natural system for point-of-care testing, such as in tap water, milk, or serum. The aptasensor is universal and expandable and holds great potential in the COVID-19 early diagnosis, environmental surveillance, food security, and other aspects.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , COVID-19 , Sistemas CRISPR-Cas , Técnicas Electroquímicas , Electrodos , Oro , Humanos , Límite de Detección , Proteínas de la Nucleocápside , SARS-CoV-2
16.
Anal Chem ; 93(49): 16646-16654, 2021 12 14.
Artículo en Inglés | MEDLINE | ID: mdl-34847324

RESUMEN

With the outbreak of COVID-19, which is fast transmitting and highly contagious, the development of rapid, highly specific, and sensitive detection kits has become a research hotspot. The existing assay methods for SARS-CoV-2 are mainly based on enzymatic reactions, which require expensive reagents, hindering popular use, especially in resource-constrained areas. Herein, we propose an aptamer-based method for the assay of SARS-CoV-2 via binding of the spike protein using functionalized biomimetic nanochannels. To get the analogous effect of human ACE2, a receptor for the spike protein, the aptamer to bind to the spike S1 protein has been first screened by a SELEX technique and then immobilized on the previously prepared nanochannels. In the presence of SARS-CoV-2, the changes in steric hindrance and charge density on the surface of the nanochannels will affect the ion transport, along with a rapid electrochemical response. Our method has been successfully applied to detect the viral particles in clinical pharyngeal swab specimens in one step without sample treatment. We expect this rapid, reagent-free, and sensitive assay method to be developed as a useful tool for diagnosing COVID-19.


Asunto(s)
COVID-19 , SARS-CoV-2 , Humanos
17.
Nanomaterials (Basel) ; 11(4)2021 Mar 25.
Artículo en Inglés | MEDLINE | ID: mdl-33806173

RESUMEN

Misuse of antibiotics has recently been considered a global issue because of its harmful effects on human health. Since conventional methods have numerous limitations, it is necessary to develop fast, simple, sensitive, and reproducible methods for the detection of antibiotics. Among numerous recently developed methods, aptasensors are fascinating because of their good specificity, sensitivity and selectivity. These kinds of biosensors combining aptamer with colorimetric applications of gold nanoparticles to recognize small molecules are becoming more popular owing to their advantageous features, for example, low cost, ease of use, on-site analysis ability using naked eye and no prerequisite for modern equipment. In this review, we have highlighted the recent advances and working principle of gold nanoparticles based colorimetric aptasensors as promising methods for antibiotics detection in different food and environmental samples (2011-2020). Furthermore, possible advantages and disadvantages have also been summarized for these methods. Finally, the recent challenges, outlook, and promising future perspectives for developing novel aptasensors are also considered.

18.
ACS Appl Mater Interfaces ; 13(14): 16118-16126, 2021 Apr 14.
Artículo en Inglés | MEDLINE | ID: mdl-33787226

RESUMEN

Diagnosis and treatment of ovarian cancer are based on intraoperative pathology and debulking surgery. The development of a novel molecular tool is significant for rapid intraoperative pathologic diagnosis, which instructs the decision-making on excision surgery and effective chemotherapy. In this work, we represent a DNA aptamer named mApoc46, which is generated from cell-SELEX by targeting patient-derived primary serous ovarian cancer (pSOC) cells. An average dissociation constant (Kd) was determined to be 0.15 ± 0.05 µM by flow cytometry. The mApoc46 aptamer displays a robust specificity to pSOC cells. Labeled with FAM, mApoc46 can selectively stain living pSOC cells in 30 min without staining commercial OC cell lines and cell lines associated with other cancers. Interestingly, FAM-mApoc46 displayed superb selectivity toward high-grade serous ovarian cancer (HG-SOC) tissues in frozen sections against low-grade SOC, ovarian borderline tumor, other nonepithelial ovarian tumors, and healthy ovarian tissue. These results lead to a potential application in the identification of OCs' histological subtypes during operation. In the patient-derived tumor xenograft NCG mice model, Cy5-labeled mApoc46 was found to accumulate at the tumor area and served as an in vivo imaging probe. The mApoc46 probe shows a robust and stable performance to visualize SOC tumors in the body. Therefore, aptamer mApoc46 holds great potential in rapid intraoperative detection, pathological diagnosis, fluorescence image-guided cancer surgery, and targeted drug delivery and therapy.


Asunto(s)
Aptámeros de Nucleótidos , Neoplasias Ováricas/diagnóstico , Animales , Femenino , Xenoinjertos , Humanos , Periodo Intraoperatorio , Ratones , Neoplasias Ováricas/diagnóstico por imagen , Técnica SELEX de Producción de Aptámeros
19.
Front Oncol ; 11: 632165, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33718215

RESUMEN

Cancer is a life-threatening concern worldwide. Sensitive and early-stage diagnostics of different cancer types can make it possible for patients to get through the best available treatment options to combat this menace. Among several new detection methods, aptamer-based biosensors (aptasensors) have recently shown promising results in terms of sensitivity, identification, or detection of either cancerous cells or the associated biomarkers. In this mini-review, we have summarized the most recent (2016-2020) developments in different approaches belonging to optical aptasensor technologies being widely employed for their simple operation, sensitivity, and early cancer diagnostics. Finally, we shed some light on limitations, advantages, and current challenges of aptasensors in clinical diagnostics, and we elaborated on some future perspectives.

20.
Nucleic Acids Res ; 48(19): 10680-10690, 2020 11 04.
Artículo en Inglés | MEDLINE | ID: mdl-33021630

RESUMEN

Circular DNA aptamers are powerful candidates for therapeutic applications given their dramatically enhanced biostability. Herein we report the first effort to evolve circular DNA aptamers that bind a human protein directly in serum, a complex biofluid. Targeting human thrombin, this strategy has led to the discovery of a circular aptamer, named CTBA4T-B1, that exhibits very high binding affinity (with a dissociation constant of 19 pM), excellent anticoagulation activity (with the half maximal inhibitory concentration of 90 pM) and high stability (with a half-life of 8 h) in human serum, highlighting the advantage of performing aptamer selection directly in the environment where the application is intended. CTBA4T-B1 is predicted to adopt a unique structural fold with a central two-tiered guanine quadruplex capped by two long stem-loops. This structural arrangement differs from all known thrombin binding linear DNA aptamers, demonstrating the added advantage of evolving aptamers from circular DNA libraries. The method described here permits the derivation of circular DNA aptamers directly in biological fluids and could potentially be adapted to generate other types of aptamers for therapeutic applications.


Asunto(s)
Aptámeros de Nucleótidos/química , ADN Circular/química , Trombina/metabolismo , Aptámeros de Nucleótidos/sangre , Aptámeros de Nucleótidos/metabolismo , ADN Circular/sangre , ADN Circular/metabolismo , G-Cuádruplex , Humanos , Unión Proteica , Trombina/química
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