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Mesoporous polydopamine with built-in plasmonic core: Traceable and NIR triggered delivery of functional proteins
摘要： Functional proteins are essential for the regulation of cellular behaviors and have found growing therapeutic uses. However, low bioavailability of active proteins to their intracellular targets has been a long-standing challenge to achieve their full potential for cell reprogramming and disease treatment. Here we report mesoporous polydopamine (mPDA) with a built-in plasmonic nanoparticle core as a multifunctional protein delivery system. The mPDA with a unique combination of large surface area, metal-chelating property, and broad-spectrum photothermal transduction allows efficient loading and near-infrared light-triggered release of functional proteins, while the plasmonic core serves as a photostable tracer and fluorescence quencher, collectively leading to real-time monitoring and active cytosolic release of model proteins. In particular, controlled delivery of cytotoxic ribonuclease A has shown excellent performance in in-vivo cancer therapy. The possibility of coating mPDA on a broad range of functional cores, thanks to its universal adhesion, provides opportunities for developing tailored delivery carriers of biologics to overcome intrinsic biological barriers.
关键词： cancer therapy,photothermal-responsive,traceable protein delivery,plasmonic nanostructures,mesoporous polydopamine
更新于2025-09-19 17:13:59
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Attenuation correction for confocal laser scanning microscopy and its application in chromatography
摘要： The applicability of confocal laser scanning microscopy is limited e.g. by attenuation of the excitation and the fluorescence emission beam. As a prerequisite for further processing and analysis of the obtained microscopic images, a new method is presented for correcting this attenuation. The correction is based on beam modeling and on a differential form of the modified Beer-Lambert law. It turns out that the intensity decay can be modeled as a double convolution of the microscopic image with the intensities of the excitation semi-beam and the emission beam. Under weak assumptions made for the intensities of the fluorescent radiation and the detected signal, formulas for the attenuation correction and the attenuation simulation are derived. The method traces back to that one published by Roerdink which is modified concerning a more realistic beam modeling, avoiding the so-called weak attenuation expansion, and considering fluorescence excitation throughout the light cone of the excitation beam. The applicability of the method is demonstrated for synthetic examples as well as microscopic images of chromatographic beads. It is shown that the new method can be successfully applied for reconstructing the true fluorophore distribution in specimens even if the microscopic images are affected by strong attenuation.
关键词： shading correction,protein distribution,chromatographic purification,Fluorescence microscopy
更新于2025-09-19 17:13:59
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Zwitterion Effect of Cow Brain Protein towards Efficiency Improvement of Dye-Sensitized Solar Cell (DSSC)
摘要： Dye-Sensitized Solar Cell (DSSC) constitutes a solar cell using natural dyes from plants that are adsorbed in semiconductors to convert solar energy into electrical energy. DSSC has relatively inexpensive fabrication costs, is easy to produce, works in visible light, and is environmentally friendly. The disadvantage of DSSC is that its e?ciency is still low compared to silicon solar cells. This low e?ciency is due to obstacles in the ?ow of electric current on DSSC. In this study, DSSC has been successfully fabricated with the deposition of clathrin protein from cow brain. The zwitterions e?ect of protein on cow brain is able to reduce resistance and increase electric current on DSSC. The zwitterions e?ect of cow brain protein that ?lls gaps or empty spaces between TiO2 particles generates acidic reactions (capturing electrons) and bases (releasing electrons); hence, proteins in the cow brain are able to function as electron bridges between TiO2 molecules and generate an increase in electric current in DSSC. The method used in this research was to deposit clathrin protein from cow brain in a porous TiO2 semiconductor with a concentration of 0%, 25%, 50%, and 75%. Tests carried out on DSSC that have been performed were X-Ray Di?ractometer (XRD) testing to determine the crystal structure formed, Fourier Transform Infrared Spectroscopy (FTIR) testing to determine the functional groups formed on DSSC, Scanning Electron Microscopy (SEM) testing to determine the surface morphological characteristics of the DSSC layer, and testing the e?ciency using AM 1.5 G solar simulator (1000 W/m2) to determine the e?ciency changes that occur in DSSC. From the XRD test results by increasing the concentration of cow brain protein in DSSC, the structure of amino acid crystals also increased and the crystal size increased with the largest crystal size of 42.25 nm at the addition of 75% of cow brain protein. FTIR test results show that the addition of cow brain protein will form functional protein-forming amino groups on DSSC. FTIR analysis shows the sharp absorption of energy by protein functional groups in the FTIR spectrum with increasing concentration of cow brain protein in DSSC. The SEM test results show that the concentration of additional molecules of protein deposited into TiO2 increases and the cavity or pore between the TiO2 molecules decreases. The reduction of cavities in the layers indicates that protein molecules ?ll cavities that exist between TiO2 molecules. From the results of testing using AM 1.5 G solar simulator (1000 W/m2), the highest e?ciency value is 1.465% with the addition of 75% brain protein concentration.
关键词： DSSC,efficiency improvement,clathrin protein,zwitterions effect,TiO2,Dye-Sensitized Solar Cell,cow brain
更新于2025-09-19 17:13:59
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White-emitting protein-metal nanocluster phosphors for highly performing bio-hybrid light-emitting diodes
摘要： This work presents a simple in-situ synthesis and stabilization of fluorescent gold nanoclusters (AuNCs) with different sizes using engineered protein scaffolds in water. The protein-AuNC hybrids show a dual emission (450 and 700 nm) with a record photoluminescence quantum yield of 20%. These features impelled us to apply them to bio-hybrid light-emitting diodes (Bio-HLEDs) as color down-converting filters or bio-phosphors. Efficient white emission (x/y CIE color coordinates of 0.31/0.29) and stabilities of >800 h were achieved. This represents a two orders of magnitude enhancement compared to the prior art. Besides the outstanding performance, the protein scaffold also infers a unique anisotropic emission character that is considered as a proof-of-concept of high interest for single-point lighting and display.
关键词： bio-phosphors,protein design,Light-emitting diodes,repeat proteins,bio-hybrid materials,metal nanocluster
更新于2025-09-19 17:13:59
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Laser microdissection-based microproteomics of the hippocampus of a rat epilepsy model reveals regional differences in protein abundances
摘要： Mesial temporal lobe epilepsy (MTLE) is a chronic neurological disorder affecting almost 40% of adult patients with epilepsy. Hippocampal sclerosis (HS) is a common histopathological abnormality found in patients with MTLE. HS is characterised by extensive neuronal loss in different hippocampus sub-regions. In this study, we used laser microdissection-based microproteomics to determine the protein abundances in different regions and layers of the hippocampus dentate gyrus (DG) in an electric stimulation rodent model which displays classical HS damage similar to that found in patients with MTLE. Our results indicate that there are differences in the proteomic profiles of different layers (granule cell and molecular), as well as different regions, of the DG (ventral and dorsal). We have identified new signalling pathways and proteins present in specific layers and regions of the DG, such as PARK7, RACK1, and connexin 31/gap junction. We also found two major signalling pathways that are common to all layers and regions: inflammation and energy metabolism. Finally, our results highlight the utility of high-throughput microproteomics and spatial-limited isolation of tissues in the study of complex disorders to fully appreciate the large biological heterogeneity present in different cell populations within the central nervous system.
关键词： microproteomics,rat epilepsy model,hippocampus,protein abundances,Laser microdissection,PARK7,energy metabolism,connexin 31/gap junction,RACK1,inflammation
更新于2025-09-19 17:13:59
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The impact of ultraviolet- and infrared-based laser microdissection technology on phosphoprotein detection in the laser microdissection-reverse phase protein array workflow
摘要： Reversible protein phosphorylation represents a key mechanism by which signals are transduced in eukaryotic cells. Dysregulated phosphorylation is also a hallmark of carcinogenesis and represents key drug targets in the precision medicine space. Thus, methods that preserve phosphoprotein integrity in the context of clinical tissue analyses are crucially important in cancer research. Here we investigated the impact of UV laser microdissection (UV LMD) and IR laser capture microdissection (IR LCM) on phosphoprotein abundance of key cancer signaling protein targets assessed by reverse-phase protein microarray (RPPA). Tumor epithelial cells from consecutive thin sections obtained from four high-grade serous ovarian cancers were harvested using either UV LMD or IR LCM methods. Phosphoprotein abundances for ten phosphoproteins that represent important drug targets were assessed by RPPA and revealed no significant differences in phosphoprotein integrity from those obtained using higher-energy UV versus the lower-energy IR laser methods.
关键词： Reverse phase protein array,Phosphoprotein,Proteomics,Laser capture microdissection,Laser microdissection
更新于2025-09-19 17:13:59
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Probing Specificity of Protein–Protein Interactions with Chiral Plasmonic Nanostructures
摘要： Protein?protein interactions (PPIs) play a pivotal role in many biological processes. Discriminating functionally important well-defined protein?protein complexes formed by specific interactions from random aggregates produced by nonspecific interactions is therefore a critical capability. While there are many techniques which enable rapid screening of binding affinities in PPIs, there is no generic spectroscopic phenomenon which provides rapid characterization of the structure of protein?protein complexes. In this study we show that chiral plasmonic fields probe the structural order and hence the level of PPI specificity in a model antibody?antigen system. Using surface-immobilized Fab′ fragments of polyclonal rabbit IgG antibodies with high specificity for bovine serum albumin (BSA), we show that chiral plasmonic fields can discriminate between a structurally anisotropic ensemble of BSA-Fab′ complexes and random ovalbumin (OVA)-Fab′ aggregates, demonstrating their potential as the basis of a useful proteomic technology for the initial rapid high-throughput screening of PPIs.
关键词： specificity,Protein?protein interactions,structural order,chiral plasmonic nanostructures,high-throughput screening
更新于2025-09-16 10:30:52
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Optical Detection of Denatured Ferritin Protein via Plasmonic Gold Nanoparticles Exposure through Aminosilane Solution
摘要： The presence of denatured proteins within a therapeutic drug product can create a series of serious adverse effects, such as mild irritation, immunogenicity, anaphylaxis, or instant death to a patient. The detection of protein degradation is complicated and expensive due to current methods associated with expensive instrumentation, reagents, and processing time. We have demonstrated here a platform for visual biosensing of denatured proteins that is fast, low cost, sensitive, and user friendly by exploiting the plasmonic properties of noble metal nanoparticles. In this study we have exposed artificially heat stressed ferritin and gold nanoparticles to 3-aminopropyl triethoxysilane, which degrades the protein by showing a systematic blue shift in the absorbance spectra of the gold nanoparticle/ferritin and aminosilane solution. This blue shift in absorbance produces a detectable visual color transition from a blue color to a purple hue. By studying the Raman spectroscopy of the gold nanoparticle/ferritin and aminosilane solution, the extent of ferritin degradation was quantified. The degradation of ferritin was again confirmed using dynamic light scattering and was attributed to the aggregation of the ferritin due to accelerated heat stress. We have successfully demonstrated a proof of concept for visually detecting ferritin from horse spleen that has experienced various levels of degradation, including due to heat stress.
关键词： ferritin,gold nanoparticles,biosensor,denatured protein,degraded protein,visual detection
更新于2025-09-16 10:30:52
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Laser switching contrast microscopy to monitor free and restricted diffusion inside the cell nucleus
摘要： A novel microscopic technique termed laser switching contrast microscopy (LSCM) allows for the imaging of the dynamics of optically switchable proteins in single cell compartments. We present an application for the monitoring of diffusive properties of single molecules of the photo-switchable fluorescent protein Dreiklang (DRK). LSCM in the cell nucleus of Chinese hamster ovary (CHO) cells cytoplasmically expressing DRK unravels quick diffusive equilibration of the DRK molecules inside the whole cytoplasm and inside the cell nucleus within seconds. The nuclear membrane is also highly permeable for DRK. Inside the nucleus entirely distinct regions are found that only partially enable diffusive protein redistribution with mean square displacement proportional to time while in other regions the mobility of the proteins seems to be restricted. After photo-switching string like patterns of light DRK molecules are observed in the cell nucleus. In addition a fraction of these DRK molecules appears immobile. The findings support recent theories of the cell interior described as a random obstacle model with an additional immobile fraction of DRK. Numerical simulations show that at different illumination intensity and different distance from the laser focus similar patterns for fluorescence recovery might be obtained in spite of strongly varying diffusion constants.
关键词： random walk,Fluorescence microscopy,diffusion constant,nanofluidics,Dreiklang,superresolution microscopy,fluorescent protein,photo-switchable molecules
更新于2025-09-16 10:30:52
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Label-free quantum dot conjugates for human protein IL-2 based on molecularly imprinted polymers
摘要： Herein, the development of a fluorescent-based sensor by combining quantum dots (QDs) with molecularly-imprinted technology (MIP), intensively optimized to generate exceptional operating features is presented. This sensor is designed to target human interleukin-2 (IL-2) in synthetic human serum. IL-2 is a regulatory protein released as a triggered response from the immune system towards an inflammation. For this purpose, cadmium telluride (CdTe) QDs are prepared with 3-mercaptopropionic acid (MPA) and modified afterwards to produce an IL-2 imprinted polymer with methacrylic acid and N,N′-methylenebis(acrylamide), upon removal of the template under optimized conditions. During IL-2 rebinding, the fluorescence intensity of CdTe@MPA QDs is quenched in a concentration dependent manner. Using surface imprinting technology, the optimal fluorescence signals yielded a linear response versus logarithm of IL-2 concentration from 35 fg/ml to 39 pg/ml, in a 1000-fold diluted synthetic human serum. The limit of detection obtained is 5.91 fg/ml, lying below the concentration levels of IL-2 with clinical interest for cancer diagnosis (9.4-19.2 pg/ml). Overall, the method presented herein is a demonstration that the combination of MIP and QDs for protein detection constitutes a powerful tool in clinical analysis, providing low cost, sensitive and quick responses. The same concept may be further extended to other proteins of interest.
关键词： interleukin-2,conjugated-QDs,protein,molecularly imprinted polymer,Quantum dots,cancer biomarker
更新于2025-09-16 10:30:52