RESUMO
The development of sensitive methods for alkaline phosphatase (ALP) activity analysis is an important analytical topic. Based on the stimulus-responsive lanthanide coordination polymer, a simple ratiometric fluorescence sensing strategy was proposed to detect ALP activity. A carbon dot (CD) doped fluorescent supramolecular lanthanide coordination polymer (CDs@Tb-GMP) was prepared with Tb3+ and the ligand guanine single nucleotide (GMP). To construct a ratiometric fluorescence biosensor, the fluorescence of Tb-GMP was used as a response signal, and the fluorescence of CDs was used as a reference signal due to its good stability. When excited at 290 nm, the polymer network Tb-GMP emits characteristic fluorescence at 545 nm, while the CDs encapsulated in the polymer network emit fluorescence at 370 nm. After adding ALP to the system, the substrate GMP can be hydrolyzed by ALP, resulting in the destruction of the polymer network. Accordingly, the fluorescence of Tb-GMP significantly decreased, while the fluorescence of CDs slightly increased due to their release from the polymer network. By comparing the relationship between the fluorescence intensity ratio of the two signals and the concentration of ALP, sensitive detection of ALP could be achieved with the linear range from 0.5 to 80 U L-1 and a detection limit of 0.13 U L-1. Furthermore, the proposed ratiometric sensing system was applied to the detection of ALP in human serum samples with desirable results, indicating potential application in clinical diagnosis.
Assuntos
Elementos da Série dos Lantanídeos , Nanocompostos , Fosfatase Alcalina , Carbono , Corantes Fluorescentes , Humanos , PolímerosRESUMO
Exosomes, as emerging "next-generation" biotherapeutics and drug delivery vectors, hold immense potential in diverse biomedical fields, ranging from drug delivery and regenerative medicine to disease diagnosis and tumor immunotherapy. However, the rapid clearance by traditional bolus injection and poor stability of exosomes restrict their clinical application. Microneedles serve as a solution that prolongs the residence time of exosomes at the administration site, thereby maintaining the drug concentration and facilitating sustained therapeutic effects. In addition, microneedles also possess the ability to maintain the stability of bioactive substances. Therefore, we introduce a microneedle patch for loading and delivering exosomes and share the methods, including isolation of exosomes, fabrication, and characterization of exosome-loaded microneedle patches. The microneedle patches were fabricated using trehalose and hyaluronic acid as the tip materials and polyvinylpyrrolidone as the backing material through a two-step casting method. The microneedles demonstrated robust mechanical strength, with tips able to withstand 2 N. Pig skin was used to simulate human skin, and the tips of microneedles completely melted within 60 s after skin puncture. The exosomes released from the microneedles exhibited morphology, particle size, marker proteins, and biological functions comparable to those of fresh exosomes, enabling dendritic cells uptake and promoting their maturation.
Assuntos
Sistemas de Liberação de Medicamentos , Exossomos , Ácido Hialurônico , Microinjeções , Agulhas , Exossomos/química , Animais , Suínos , Sistemas de Liberação de Medicamentos/métodos , Sistemas de Liberação de Medicamentos/instrumentação , Microinjeções/métodos , Microinjeções/instrumentação , Ácido Hialurônico/química , Humanos , Povidona/química , Adesivo Transdérmico , Trealose/químicaRESUMO
In the past decades, various nanomaterials with unique properties have been explored for bioapplications. Meanwhile, aptamers, generated from the systematic evolution of ligands by exponential enrichment technology, are becoming an indispensable element in the design of functional nanomaterials because of their small size, high stability, and convenient modification, especially endowing nanomaterials with recognition capability to specific targets. Therefore, the incorporation of aptamers into nanomaterials offers an unprecedented opportunity in the research fields of diagnostics and therapeutics. Here, we focus on recent advances in aptamer-embedded nanomaterials for bioapplications. First, we briefly introduce the properties of nanomaterials that can be functionalized with aptamers. Then, the applications of aptamer-embedded nanomaterials in cellular analysis, imaging, targeted drug delivery, gene editing, and cancer diagnosis/therapy are discussed. Finally, we provide some perspectives on the challenges and opportunities that have arisen from this promising area.
Assuntos
Aptâmeros de Nucleotídeos/química , Ácidos Nucleicos Imobilizados/química , Nanopartículas Metálicas/química , Neoplasias/diagnóstico por imagem , Neoplasias/tratamento farmacológico , Animais , Antineoplásicos/uso terapêutico , Sequência de Bases , Portadores de Fármacos/química , Humanos , Hidrogéis/química , Lipossomos/química , Estruturas Metalorgânicas/química , Micelas , NanoporosRESUMO
We report an ultrasensitive electrochemical immunosensor designed for the detection of protein biomarkers using horseradish peroxidase (HRP)-loaded silica-poly(acrylic acid) brushes (SiO2-SPAABs) as labels. HRP could be efficiently and stably accommodated in the three-dimensional architecture of the SiO2-SPAABs and the SiO2-SPAABs-HRP exhibited high catalytic performance towards o-phenylenediamine (OPD) oxidation in the presence of H2O2, which resulted in significant differential pulse voltammetric (DPV) response change and color change. Using human IgG (HIgG) as a model analyte, a sandwich-type immunosensor was constructed. In particular, graphene oxide (GO) and SiO2-SPAABs-HRP were used to immobilize capture antibody (Ab1) and bind a layer of detection antibody (Ab2), respectively. The current biosensor exhibited a good linear response of HIgG from 100pg/mL to 100µg/mL with a detection limit of 50pg/mL (S/N=5). The sensitivity was 6.70-fold higher than the conventional enzyme-linked immunosorbent assays. The immunosensor results were validated through the detection of HIgG in serum samples.
Assuntos
Biomarcadores/sangue , Técnicas Biossensoriais , Técnicas Eletroquímicas , Imunoglobulina G/isolamento & purificação , Resinas Acrílicas/química , Anticorpos/química , Anticorpos/imunologia , Biomarcadores/química , Ouro , Grafite/química , Peroxidase do Rábano Silvestre/química , Humanos , Peróxido de Hidrogênio/química , Imunoensaio , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Fenilenodiaminas/química , Dióxido de Silício/químicaRESUMO
The ultrasensitive monitoring of glucose with a fast and accurate method is significant in potential therapeutics and optimizes protein biosynthesis. Incorporation of enzyme into matrix is considered as promising candidates for constructing highly sensitive glucose-responsive systems. In this study, three-dimensional poly(acrylic acid) brushes-nanospherical silica (PAA-nano silica) with high amplification capability and stability were used to covalently immobilize bienzymes for cascade enzymatic catalysis. The major advantages of PAA-nano silica-bienzyme co-incorporation is that the enzymes are proximity distribution, and such close confinement both minimized the diffusion of intermediates among the enzymes in the consecutive reaction and improve the utilization efficiency of enzymes, thereby enhancing the overall reaction efficiency and specificity. Thus, this present bienzymatic biosensor shows robust signal amplification and ultrasensitivity of glucose-responsive properties with a detection limit of 0.04µM.
Assuntos
Resinas Acrílicas/química , Biocatálise , Glucose Oxidase/metabolismo , Glucose/análise , Peroxidase do Rábano Silvestre/metabolismo , Nanopartículas/química , Dióxido de Silício/química , Técnicas Biossensoriais , Difusão , Glucose/química , Glucose Oxidase/química , Peroxidase do Rábano Silvestre/química , Peróxido de Hidrogênio/química , Modelos Moleculares , Conformação MolecularRESUMO
A novel colorimetric immunoassay was designed for the sensitive detection of carbohydrate antigen 125 (CA125). The success of this immunoassay relies on the use of hollow polydopamine-gold nanoparticles (PDA-Au) for signal amplification to achieve sensitive nonenzymatic colorimetric detection. In particular, PDA-Au was used as a stable and sensitive label and aminated-Fe3O4 magnetic nanoparticles (Fe3O4 NPs) were employed to immobilize capture antibody (Ab1) and acted as a separable immunosensing probe. PDA-Au exhibited high catalytic performance towards p-nitrophenol reduction and thus resulted in significant color change and UV/vis signal variations. The immunoassay was performed based on sandwich protocol. As compared to pure Au nanoparticles, the signal amplification and sensitivity of PDA-Au-based assay was significantly improved. For instance, the dynamic range of the developed colorimetric immunoassay for CA125 was 0.1-100 U/mL with a detection limit of 0.1 U/mL at S/N=3. In addition, this immunoassay was also tested for the analysis of clinical serum samples, which demonstrated its potential for practical diagnostic applications.
Assuntos
Anticorpos Imobilizados/química , Técnicas Biossensoriais/métodos , Antígeno Ca-125/sangue , Ouro/química , Imunoensaio/métodos , Indóis/química , Nanopartículas/química , Polímeros/química , Colorimetria/métodos , Humanos , Limite de Detecção , Nanopartículas de Magnetita/química , Nanopartículas de Magnetita/ultraestrutura , Nanopartículas/ultraestruturaRESUMO
A new kind of signal amplification strategy based on ferrocene (Fc) incorporated polystyrene spheres (PS-Fc) was proposed. The synthesized PS-Fc displayed narrow size distribution and good stability. PS-Fc was applied as label to develop immunosensors for prostate specific antigen (PSA) after the typical sandwich immunoreaction by linking anti-PSA antibody (Ab2) onto PS-Fc. After the fabrication of the immunosensor, tetrahydrofuran (THF) was dropped to dissolve PS and release the contained Fc for the following stripping voltammetric detection. PS-Fc as a new electrochemical label prevented the leakage of Fc and greatly amplified the immunosensor signal. In addition, the good biocompatibility of PS could maintain the bioactivity of the antibodies. The response current was linear to the logarithm of PSA concentration in the range from 0.01 ng mL(-1) to 20 ng mL(-1) with a detection limit of 1 pg mL(-1). The immunosensor results were validated through the detection of PSA in serum samples with satisfactory results.
Assuntos
Técnicas Eletroquímicas , Compostos Ferrosos/química , Imunoensaio , Poliestirenos/química , Antígeno Prostático Específico/sangue , Anticorpos/imunologia , Compostos Ferrosos/metabolismo , Grafite/química , Humanos , Masculino , Metalocenos , Óxidos/química , Antígeno Prostático Específico/imunologiaRESUMO
We present the synthesis and multifunctional utilization of core-shell Fe3O4 polydopamine nanoparticles (Fe3O4@PDA NPs) to serve as the enabling platform for a range of applications including responsive drug delivery, recyclable catalyst support, and adsorbent. Magnetite Fe3O4 NPs formed in a one-pot process by the hydrothermal approach were coated with a polydopamine shell layer of ~20 nm in thickness. The as prepared Fe3O4@PDA NPs were used for the controlled drug release in a pH-sensitive manner via reversible bonding between catechol and boronic acid groups of PDA and the anticancer drug bortezomib (BTZ), respectively. The facile deposition of Au NPs atop Fe3O4@PDA NPs was achieved by utilizing PDA as both the reducing agent and the coupling agent. The nanocatalysts exhibited high catalytic performance for the reduction of o-nitrophenol. Furthermore, the recovery and reuse of the catalyst was demonstrated 10 times without any detectible loss in activity. Finally, the PDA layers were converted into carbon to obtain Fe3O4@C and used as an adsorbent for the removal of Rhodamine B from an aqueous solution. The synergistic combination of unique features of PDA and magnetic nanoparticles establishes these core-shell NPs as a versatile platform for multiple applications.
Assuntos
Carbono/química , Portadores de Fármacos/química , Óxido Ferroso-Férrico/química , Indóis/química , Nanopartículas de Magnetita/química , Polímeros/química , Adsorção , Ácidos Borônicos/química , Bortezomib , Catálise , Tamanho da Partícula , Pirazinas/química , Rodaminas/química , Água/químicaRESUMO
Conducting polymer film was prepared by electrochemical polymerization of aniline. Multiwalled carbon nanotubes (MWNTs) were treated with a mixture of concentrated sulfuric and nitric acid to introduce carboxylic acid groups to the nanotubes. By using the layer-by-layer method, homogeneous and stable MWNTs and polyaniline (PANI) multilayer films were alternately assembled on glassy carbon (GC) electrodes. Conducting polymer of PANI had three main functions: (i) excellent antiinterference ability, (ii) protection ability in favor of increasing the amount of the MWNTs immobilized on GC electrodes, and (iii) superior transducing ability. The protection effect of PANI film and the electrostatic interaction between positively charged PANI and negatively charged MWNTs both attributed to immobilizing abundant MWNTs stably, thereby enhancing the catalytic activity. The layer-by-layer assembled MWNTs and PANI-modified GC electrodes offered a significant decrease in the overvoltage for the H2O2 and were shown to be excellent amperometric sensors for H2O2 from +0.2V over a wide range of concentrations. As an application example, by linking choline oxidase (CHOD), an amplified biosensor toward choline was prepared. The choline biosensor exhibited a linear response range of 1x10(-6) to 2x10(-3) M with a correlation coefficient of 0.997, and the response time and detection limit (S/N=3) were determined to be 3 s and 0.3 microM, respectively. The antiinterference biosensor displays a rapid response and an expanded linear response range as well as excellent reproducibility and stability.