RESUMEN
We have successfully developed a target-responsive aptamer cross-linked hydrogel for the visual detection of glucose, an important biomedical analyte. In this work, the glucose-responsive hydrogel was prepared using the target aptamer and its two short complementary DNA strands grafted onto a linear polyacrylamide chain as cross-linkers. Gold nanoparticles (AuNPs) modified with thiol-PEG were encapsulated in the gel and used as the output signal for visible detection. The complex of glucose and its ligand of boronic acid derivatives (Shinkai's receptor) can bind with the aptamer to disrupt the hydrogel, leading to the release of AuNPs with a distinct red colour in the supernatant. By this method glucose can be detected with the naked eye, and the sensor has a detection limit of 0.44 mM in buffer with the help of UV-Vis spectrophotometry. Furthermore, glucose spiked in 50% urine and 30% serum could also be detected respectively with the naked eye, and glucose was quantitatively detected in 50% urine. The hydrogel system provides a non-enzymatic and visual method for glucose detection, and offers promising applications in biotechnology and biomedicine.
Asunto(s)
Técnicas Biosensibles , ADN/química , Glucosa/análisis , Hidrogeles , Nanopartículas del Metal , Aptámeros de Nucleótidos/química , OroRESUMEN
It is demonstrated that DNA can be used to control the synthesis of silver nanoplates with different morphologies using spherical silver seeds. UV-vis spectroscopy, transmission electron microscopy, scanning electron microscopy, X-ray photoelectron spectroscopy, and Raman spectroscopy are used to characterize the synthesized nanoparticles. Silver nanoprisms are encoded by poly C and poly G, while silver flower bouquets and silver nanodiscs are synthesized using poly A and poly T, respectively. The length of DNA is found to have little effect on the morphology of silver nanoparticles. Moreover, the synthesized silver nanoplates are found to have high surface enhanced Raman scattering enhancement ability, good antibacterial activity, and good biocompatibility. These discoveries will broaden the application of DNA in nanoscience and will provide a new platform to investigate the interaction between DNA sequences and silver nanoparticles.
Asunto(s)
ADN/química , Nanopartículas del Metal/química , Plata/química , Secuencia de Bases , Células HeLa , Humanos , Cinética , Nanopartículas del Metal/ultraestructura , Espectroscopía de Fotoelectrones , Espectrofotometría UltravioletaRESUMEN
Fabrication of organic thin films with highly dispersed inorganic nanoparticles is a very challenging topic. In this work, a new approach that combines electron-induced molecular self-assembly with simultaneous nanoparticle formation by room temperature electron reduction was developed to prepare peptide thin films with highly dispersed noble metal nanoparticles. Argon glow discharge was employed as the resource of electrons. The peptide motif KLVFF (Aß16-20) self-assembled into two-dimensional membranes under the influence of hydrated electrons, while the metal ions in solution can be simultaneously reduced by electrons to form nanoparticles. Our TEM imaging reveals that metal nanoparticles were well-distributed in the resulting peptide thin films. Our results also suggest that the size of metal nanoparticles can be tuned by varying the initial concentration of the metal ion. This simple approach can be viewed as a promising strategy to create hybrid thin films that integrate functional inorganics into biomolecule scaffolds.
Asunto(s)
Nanopartículas del Metal/química , Péptidos/química , Argón/química , Oro/química , Microscopía de Fuerza Atómica , Propiedades de SuperficieRESUMEN
A new method based on a functional DNA crosslinked hydrogel as a target-responsive unit and gold nanorods (AuNRs) as a multicolor signal readout circuit was developed for the sensitive and visual detection of different targets. The color variation of the AuNR solution was correlated with the concentration of the target. This system can be extended to detect various targets by designing the corresponding target-responsive DNA hydrogels.
Asunto(s)
Colorimetría/métodos , ADN/química , Oro/química , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Nanotubos/químicaRESUMEN
Aflatoxin B1 (AFB1), as the secondary metabolite of molds, is the most predominant and toxic mycotoxin that seriously threatens the health of humans and animals. In this work, an AFB1-responsive hydrogel was synthesized for highly sensitive and portable detection of AFB1. The AFB1-responsive hydrogel was prepared using an AFB1 aptamer and its two short complementary DNA strands as cross-linkers. For visual detection of AFB1, the hydrogel is preloaded with gold nanoparticles (AuNPs). Upon introduction of AFB1, the AFB1 aptamer binds with AFB1, leading to the disruption of the hydrogel and release of the AuNPs with a distinct color change of the supernatant from colorless to red. In order to lower the detection limit and extend the method to quantitative analysis, a distance-readout volumetric bar chart chip (V-chip) was combined with an AFB1-responsive hydrogel preloaded with platinum nanoparticles (PtNPs). In the presence of AFB1, the hydrogel collapses and releases PtNPs which can catalyze the decomposition of H2O2 to generate O2. The increasing gas pressure moves a red ink bar in the V-chip and provides a quantitative relationship between the distance and the concentration of AFB1. The method was applied for detection of AFB1 in beer, with a detection limit of 1.77 nM (0.55 ppb) where an immunoaffinity column (IAC) of AFB1 was used to cleanup and pre-concentrate the sample, which satisfies the testing requirement of 2.0 ppb set by the European Union. The combination of an AFB1-responsive hydrogel with a distance-based readout V-chip offers a user-friendly POCT device, which has great potential for rapid, portable, selective, and quantitative detection of AFB1 in real samples to ensure food safety and avoid subsequent economic losses.
Asunto(s)
Aflatoxina B1/análisis , Carcinógenos Ambientales/análisis , Hidrogeles/química , Dispositivos Laboratorio en un Chip , Modelos Químicos , Aptámeros de Nucleótidos/química , ADN Complementario/química , Estudios de Factibilidad , Inspección de Alimentos/instrumentación , Oro/química , Ácidos Nucleicos Inmovilizados/química , Límite de Detección , Nanopartículas del Metal/química , Platino (Metal)/químicaRESUMEN
There is considerable demand for sensitive, selective, and portable detection of disease-associated proteins, particularly in clinical practice and diagnostic applications. Portable devices are highly desired for detection of disease biomarkers in daily life due to the advantages of being simple, rapid, user-friendly, and low-cost. Herein we report an enzyme-encapsulated liposome-linked immunosorbent assay for sensitive detection of proteins using personal glucose meters (PGM) for portable quantitative readout. Liposomes encapsulating a large amount of amyloglucosidase or invertase are surface-coated with recognition elements such as aptamers or antibodies for target recognition. By translating molecular recognition signal into a large amount of glucose with the encapsulated enzyme, disease biomarkers such as thrombin or C-reactive protein (CRP) can be quantitatively detected by a PGM with a high detection limit of 1.8 or 0.30 nM, respectively. With the advantages of portability, ease of use, and low-cost, the method reported here has potential for portable and quantitative detection of various targets for different POC testing scenarios, such as rapid diagnosis in clinic offices, health monitoring at the bedside, and chemical/biochemical safety control in the field.