RESUMEN
Squamous cell carcinoma (SCC) is a common nonmelanoma skin cancer. Radiotherapy plays an integral role in treating SCC due to its characteristics, such as diminished intercellular adhesion, heightened cell migration and invasion capabilities, and immune evasion. These problems lead to inaccurate tumor boundary positioning and radiotherapy tolerance in SCC treatment. Thus, accurate localization and enhanced radiotherapy sensitivity are imperative for effective SCC treatment. To address the existing limitations in SCC therapy, we developed monoglyceride solid lipid nanoparticles (MG SLNs) and enveloped them with the A431 cell membrane (A431 CM) to create A431@MG. The characterization results showed that A431@MG was spherical. Furthermore, A431@MG had specific targeting for A431 cells. In A431 tumor-bearing mice, A431@MG demonstrated prolonged accumulation within tumors, ensuring precise boundary localization of SCC. We further advanced the approach by preparing MG SLNs encapsulating 5-aminolevulinic acid methyl ester (MLA) and desferrioxamine (DFO) with an A431 CM coating to yield A431@MG-MLA/DFO. Several studies have revealed that DFO effectively reduced iron content, impeding protoporphyrin IX (PpIX) biotransformation and promoting PpIX accumulation. Simultaneously, MLA was metabolized into PpIX upon cellular entry. During radiotherapy, the heightened PpIX levels enhanced reactive oxygen species (ROS) generation, inducing DNA and mitochondrial damage and leading to cell apoptosis. In A431 tumor-bearing mice, the A431@MG-MLA/DFO group exhibited notable radiotherapy sensitization, displaying superior tumor growth inhibition. Combining A431@MG-MLA/DFO with radiotherapy significantly improved anticancer efficacy, highlighting its potential to serve as an integrated diagnostic and therapeutic strategy for SCC.
Asunto(s)
Carcinoma de Células Escamosas , Membrana Celular , Nanopartículas , Fármacos Sensibilizantes a Radiaciones , Neoplasias Cutáneas , Animales , Ratones , Nanopartículas/química , Humanos , Línea Celular Tumoral , Neoplasias Cutáneas/tratamiento farmacológico , Neoplasias Cutáneas/patología , Carcinoma de Células Escamosas/tratamiento farmacológico , Carcinoma de Células Escamosas/radioterapia , Fármacos Sensibilizantes a Radiaciones/química , Fármacos Sensibilizantes a Radiaciones/farmacología , Fármacos Sensibilizantes a Radiaciones/administración & dosificación , Membrana Celular/metabolismo , Ácido Aminolevulínico/química , Ácido Aminolevulínico/farmacología , Ácido Aminolevulínico/administración & dosificación , Lípidos/química , Ensayos Antitumor por Modelo de Xenoinjerto , Deferoxamina/química , Deferoxamina/farmacología , Ratones Desnudos , Femenino , Ratones Endogámicos BALB C , Especies Reactivas de Oxígeno/metabolismo , Apoptosis/efectos de los fármacos , LiposomasRESUMEN
Single biofilm biomimetic nanodrug delivery systems based on single cell membranes, such as erythrocytes and cancer cells, have immune evasion ability, good biocompatibility, prolonged blood circulation, and high tumor targeting. Because of the different characteristics and functions of each single cell membrane, more researchers are using various hybrid cell membranes according to their specific needs. This review focuses on several different types of biomimetic nanodrug-delivery systems based on composite biofilms and looks forward to the challenges and possible development directions of biomimetic nanodrug-delivery systems based on composite biofilms to provide reference and ideas for future research.
Asunto(s)
Antineoplásicos , Biopelículas , Biomimética , Sistemas de Liberación de Medicamentos , Neoplasias , Humanos , Neoplasias/tratamiento farmacológico , Biopelículas/efectos de los fármacos , Biomimética/métodos , Antineoplásicos/administración & dosificación , Antineoplásicos/farmacología , Materiales Biomiméticos/química , Animales , Portadores de Fármacos/químicaRESUMEN
Based on the examination of the basic properties, the solvent extraction process (SEP) was applied with high efficiency in the extraction of bitumen from Indonesian oil sands. To separate the oil sands, different organic solvents were first screened, and the extraction effects were analyzed to select a suitable solvent. Then, the effects of operating conditions on the extraction rate of bitumen were investigated. Finally, the compositions and structures of the bitumen obtained under suitable conditions were analyzed. The results showed that the Indonesian oil sands were oil-wet oil sands with a bitumen content of 24.93%, containing a large number of asphaltenes and resins with high polarity and complex structures. The separation performance was affected by different organic solvents and operating conditions. It was shown that the closer the structure and polarity of the selected solvent is to the solute, the better the extraction effect. The extraction rate of bitumen reached 18.55% when toluene was used as the extraction solvent under the operating conditions of V (solvent):m (oil sands) 3:1, temperature 40 °C, stirring velocity 300 r/min, time 30 min. The method could also be applied to the separation of other oil-wet oil sands. The compositions and structures of bitumen can guide the separation and comprehensive use of industrial oil sands.
Asunto(s)
Hidrocarburos , Yacimiento de Petróleo y Gas , Indonesia , Hidrocarburos/química , Solventes , AlbertaRESUMEN
An efficient synthesis of a variety of 1,2-disubstituted-5,6-dihydropyrrolo[2,1-α]isoquinoline derivatives via an acid-promoted cyclization reaction between 1,2,3,4-tetrahydroisoquinoline (THIQ) and substituted α,ß-unsaturated aldehyde derivatives is reported. This cycloaddition allows access to structurally diverse multisubstituted dihydropyrrolo[2,1-α]isoquinolines in moderate to good yields, which was the core scaffold of marine natural alkaloid lamellarins.
RESUMEN
An efficient synthesis of a variety of [1,2,3]triazolo-[1,5-a]quinoxalin-4(5H)-ones via a [3 + 2] cyclization reaction by photoredox catalysis between quinoxalinones and hypervalent iodine(III) reagents is reported. A range of quinoxalinones and hypervalent iodine(III) reagents were tolerated well. This cyclization reaction allows access to structurally diverse [1,2,3]triazolo-[1,5-a]quinoxalin-4(5H)-ones in moderate to good yields.
RESUMEN
The dynamic binding status between the thrombin and its G-quadruplex aptamers and the stability of its interaction partners were probed using our previously established fluorescence-coupled capillary electrophoresis method. A 29-nucleic acid thrombin binding aptamer was chosen as a model to study its binding affinity with the thrombin ligand. First, the effects of the cations on the formation of G-quadruplex from unstructured 29-nucleic acid thrombin binding aptamer were examined. Second, the rapid binding kinetics between the thrombin and 6-carboxyfluorescein labeled G-quadruplex aptamer was measured. Third, the stability of G-quadruplex aptamer-thrombin complex was also examined in the presence of the interfering species. Remarkably, it was found that the complementary strand of 29-nucleic acid thrombin binding aptamer could compete with G-quadruplex aptamer and thus disassociated the G-quadruplex structure into an unstructured aptamer. These data suggest that our in-house established fluorescence-coupled capillary electrophoresis assay could be applied to binding studies of the G-quadruplex aptamers, thrombin, and their ligands, while overcoming the complicated and costly approaches currently available.
Asunto(s)
Aptámeros de Nucleótidos , Electroforesis Capilar , G-Cuádruplex , Trombina/química , Cationes , Fluorescencia , LigandosRESUMEN
We present a molecular characterization of metal-affinity driven self-assembly between CdSe-ZnS quantum dots and a series of hexahistidine peptides with different charges. In particular, we uti- lized fluorescence coupled capillary electrophoresis to test the self-assembly process of quantum dots with peptides in solution. Four peptides with different charges can be efficiently separated by fluorescence coupled capillary electrophoresis. The migration time appeared to be influenced by the charges of the peptide. In addition, the kinetics of self-assembly process of quantum dots with one of the peptides manifested a bi-phasic kinetics followed by a saturating stage. This work revealed that there exist two types of binding sites on the surface of quantum dots for peptide 1: one type termed "high priority" binding site and a "low priority" site which is occupied after the first binding sites are fully occupied. The total self-assembly process finishes in solution within 80 s. Our work represents the systematic investigation of the details of self-assembly kinetics utilizing high-resolution fluorescence coupled capillary electrophoresis. The charge effect of peptide coating quantum dots provides a new way of preparing bioprobes.
Asunto(s)
Cristalización/métodos , Electroforesis Capilar/métodos , Mediciones Luminiscentes/métodos , Péptidos , Puntos Cuánticos , Espectrometría de Fluorescencia/métodos , Ensayo de Materiales/métodos , Electricidad EstáticaRESUMEN
Nanoparticles provide significantly enhanced binding characteristics. However, fast online probing of the self-assembly process remains hard to achieve in practice. Herein, we report a fluorescence coupled CE method for probing the self-assembly events between quantum dots (QDs) and engineered Jumonji domain-containing protein 6 (Jmjd6) enzyme. QDs and Jmjd6 were sequentially injected into the capillary, where the self-assembly took place in a nanoliter scale. In particular, we showed that the Jmjd6/QD ratio, the interval time, and the injection volume had a great effect on the online self-assembly. The current approach may allow for a better understanding of QDs-enzyme self-assembly and enzymatic activity detection.
Asunto(s)
Electroforesis Capilar/métodos , Histona Demetilasas con Dominio de Jumonji/metabolismo , Puntos Cuánticos/metabolismo , Proteínas Recombinantes/metabolismo , Histona Demetilasas con Dominio de Jumonji/química , Histona Demetilasas con Dominio de Jumonji/genética , Puntos Cuánticos/química , Proteínas Recombinantes/química , Proteínas Recombinantes/genéticaRESUMEN
Herein, we report a technique for detecting the fast binding of antibody-peptide inside a capillary. Anti-HA was mixed and interacted with FAM-labeled HA tag (FAM-E4 ) inside the capillary. Fluorescence coupled capillary electrophoresis (CE-FL) was employed to measure and record the binding process. The efficiency of the antibody-peptide binding on in-capillary assays was found to be affected by the molar ratio. Furthermore, the stability of anti-HA-FAM-E4 complex was investigated as well. The results indicated that E4 YPYDVPDYA (E4) or TAMRA-E4 YPYDVPDYA (TAMRA-E4) had the same binding priorities with anti-HA. The addition of excess E4 or TAMRA-E4 could lead to partial dissociation of the complex and take a two-step mechanism including dissociation and association. This method can be applied to detect a wide range of biomolecular interactions.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Electroforesis Capilar/métodos , Péptidos/inmunología , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/química , Anticuerpos Monoclonales/aislamiento & purificación , Sitios de Unión de Anticuerpos , Fluorescencia , Hibridomas , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Péptidos/química , Péptidos/aislamiento & purificaciónRESUMEN
HAT tag, a natural His affinity tag, is one of the most popular fusion tags. However, HAT tag containing three positive charges limited its self-assembly with quantum dots (QDs). Herein, ATTO 590-labeled HAT peptide was synthesized to self-assemble with QDs inside the capillary. QDs and ATTO 590-labeled HAT peptide were sequentially injected into the capillary and probed by fluorescence-coupled CE (CE-FL), showing an obvious Förster resonance energy transfer signal. Online self-assembly, separation, and detection were achieved within 10 min. CE-FL further revealed that imidazole and H6 G6 peptide could partially outcompete with HAT tag on the QDs surface inside the capillary. The displacement intermediates were separated clearly using CE-FL. Our study demonstrates the power of online CE-FL in analyzing the binding interaction between ligands containing positive charges and QDs.
RESUMEN
A new method using fluorescence coupled capillary electrophoresis (CE-FL) for monitoring self-assembly and proteolytic cleavage of hexahistidine peptide capped quantum dots (QDs) inside a capillary has been developed in this report. QDs and the ATTO 590-labeled hexahistidine peptide (H6-ATTO) were injected into a capillary, sequentially. Their self-assembly inside the capillary was driven by a metal-affinity force which yielded a new fluorescence signal due to Förster resonance energy transfer (FRET). The highly efficient separation of fluorescent complexes and the FRET process were analyzed using CE-FL. The self-assembly of QDs and biomolecules was found to effectively take place inside the capillary. The kinetics of the assembly was monitored by CE-FL, and the approach was extended to the study of proteolytic cleavage of surface conjugated peptides. Being the first in-depth analysis of in-capillary nanoparticle-biomolecule assembly, the novel approach reported here provides inspiration to the development of QD-based FRET probes for biomedical applications.
Asunto(s)
Electroforesis Capilar , Histidina/química , Proteolisis , Puntos Cuánticos , FluorescenciaRESUMEN
Herein, we designed four peptides appended with different numbers of histidine (Hisn -peptide). We launched a systematic investigation on quantum dots (QDs) and Hisn -peptide self-assembly in solution using fluorescence coupled CE (CE-FL). The results indicated that CE-FL was a powerful method to probe how ligands interaction on the surface of nanoparticles. The self-assembly of QDs and peptide was determined by the numbers of histidine. We also observed that longer polyhistidine tags (n ≤ 6) could improve the self-assembly efficiency. Furthermore, the formation and separation of QD-peptide assembly were also studied by CE-FL inside a capillary. The total time for the mixing, self-assembly, separation, and detection was less than 10 min. Our method greatly expands the application of CE-FL in QDs-based biolabeling and bioanalysis.
Asunto(s)
Electroforesis Capilar/métodos , Histidina/química , Péptidos/química , Puntos Cuánticos/químicaRESUMEN
As a vast number of novel materials in particular inorganic nanoparticles have been invented and introduced to all aspects of life, public concerns about how they might affect our ecosystem and human life continue to arise. Such incertitude roots at a fundamental question of how inorganic nanoparticles self-assemble with biomolecules in solution. Various techniques have been developed to probe the interaction between particles and biomolecules, but very few if any can provide advantages of both rapid and convenient. Herein, we report a systematic investigation on quantum dots (QDs) and protein self-assembly inside a capillary. QDs and protein were injected to a capillary one after another. They were mixed inside the capillary when a high voltage was applied. Online separation and detection were then achieved. This new method can also be used to study the self-assembly kinetics of QDs and protein using the Hill equation, the KD value for the self-assembly of QDs and protein was calculated to be 8.8 µM. The obtained results were compared with the previous out of-capillary method and confirmed the effectiveness of the present method.
Asunto(s)
Electroforesis Capilar/instrumentación , Puntos Cuánticos/química , Albúmina Sérica Bovina/química , Animales , Bovinos , Fluorescencia , CinéticaRESUMEN
In this report, fluorescence detection coupled capillary electrophoresis (CE-FL) was used to detect Protein A. Antibody was first labeled with Cy5 and then mixed with quantum dots (QDs) to form QDs-antibody bioprobe. Further, we observed fluorescence resonance energy transfer (FRET) from QDs donor to Cy5 acceptor. The bioprobe was formed and brought QDs and Cy5 close enough to allow FRET to occur. After adding protein A, the FRET system was broken and caused the FRET signal to decrease. Thus, a new method for the determination of protein A was proposed based on the FRET signal changes. This study provides a new trail of thought for the detection of protein.
Asunto(s)
Anticuerpos/química , Técnicas Biosensibles , Electroforesis Capilar , Transferencia Resonante de Energía de Fluorescencia , Puntos Cuánticos/química , Proteína Estafilocócica A/química , Anticuerpos/metabolismo , Carbocianinas/química , Inmunoglobulina G/química , Inmunoglobulina G/metabolismo , Proteína Estafilocócica A/metabolismoRESUMEN
Natural antibodies adopt multivalent constructs to effect superior binding affinities with their antigens. Notwithstanding that the structure of antibodies have been well understood, how antibodies harness multivalency effect to achieve superior binding affinity toward the antigens still remains unclear. Such investigation is often hampered by the difficulty in resolving receptor-ligand complexes with different stoichiometries in the binding solution, especially when the ligand is a small molecule or a short peptide. Here we employed a unique anti-FLAG(™) mAb M2 and FLAG(™) peptides system, together with fluorescence detection coupled capillary electrophoresis (CE-FL) to reveal how M2 achieves exceptional high affinity with FLAG peptides through multivalency. Complexation of fluorescently labeled FLAG(™) peptides with M2 leads to a pronounced mobility shift of the fluorescent peak in CE. Remarkably, CE-FL rendered a base-line separation of 1:1 and 1:2 M2-FLAG(™) complexes, allowing the quantification of different M2-FLAG(™) species. The quantitative analysis leads to a detailed dissection of the first (functional affinity) and second binding affinities (intrinsic affinity) between M2 and FLAG1 peptide. The marked difference (10(3)-10(4) fold) between these two affinities indicated that multivalency effect plays a key role for M2 to achieve highly efficient binding to FLAG(™) peptides.
Asunto(s)
Anticuerpos Monoclonales/química , Afinidad de Anticuerpos , Antígenos/química , Electroforesis Capilar/métodos , Oligopéptidos/química , Animales , Anticuerpos Monoclonales/inmunología , Especificidad de Anticuerpos , Antígenos/inmunología , Colorantes Fluorescentes , Cinética , Ligandos , Ratones , Oligopéptidos/síntesis química , Oligopéptidos/inmunología , Técnicas de Síntesis en Fase Sólida , Coloración y Etiquetado , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
In this report, the use of fluorescence detection coupled capillary electrophoresis (CE-FL) allowed us to fully characterize the antigen-antibody interaction. CE-FL allowed separation of unbound quantum dots (QDs) and ligand bound QDs and also revealed an ordered assembly of biomolecules on QDs. Further, we observed FRET from QDs donor to DyLight acceptor, which were covalently conjugated with human IgG and goat anti-human IgG, respectively. The immunocomplex was formed and the mutual affinity of the antigen and antibody brought QDs and DyLight close enough to allow FRET to occur. This novel CE-based technique can be easily extended to other FRET systems based on QDs and may have potential application in the detection of antibodies.
Asunto(s)
Complejo Antígeno-Anticuerpo/química , Antígenos/inmunología , Electroforesis Capilar , Colorantes Fluorescentes/química , Inmunoglobulina G/inmunología , Animales , Transferencia Resonante de Energía de Fluorescencia , Cabras , Humanos , Inmunoglobulina G/química , Puntos Cuánticos/químicaRESUMEN
Engineered peptide ligands with exceptionally high affinity for metal can self-assemble with nanoparticles in biological fluids. A high-affinity dendrimeric peptide ligand for CdSe-ZnS quantum dots (QDs) exhibited very fast association kinetics with QDs and reached equilibrium within 2 s. Here, we have combined a droplet-based microfluidic device with fluorescence detection based on Förster resonance energy transfer (FRET) to provide subsecond resolution in dissecting this fast self-assembly kinetics in solution. This work represents the first application of microfluidic devices to ligand-particle assembly for the measurement of fast assembly kinetics in solution.