Fluorescence Spectroscopic Techniques Research Articles

Myricetin and dihydromyricetin as JNK1 inhibitors protect against ROS‐mediated oxidative stress and apoptosis

Myricetin and dihydromyricetin as JNK1 inhibitors protect against ROS‐mediated oxidative stress and apoptosis

A new water−soluble naphthalene diimide as a highly selective fluorescent chemosensor for Cu(II) ion: Synthesis, DFT calculations, photophysical and electrochemical properties

The highly selective, sensitive, and water-soluble fluorescent sensors are desideratum for optoelectronic, environmental, biological, and biomedical applications. An innovative fluorescence chemosensor, naphthalene diimide (3) with metal binding sites, was designed, synthesized and characterized. The chemosensor's optical, electrochemical, spectroelectrochemical, and morphological properties were investigated, and then the density functional theory (DFT) simulations were conducted. The selectivity of 3 for the metals Cu(II), Ag(I), Hg(II), Mg(II), Fe(III), Ca(II), Co(II), Zn(II), Pb(II) and Cd(II) were investigated through UV–visible and fluorescence spectroscopy techniques. 3 was highly selective and sensitive toward Cu(II) metal ions. The chemosensor can detect Cu(II) in water with a detection limit of 1.11 μM, which is lower than the WHO standard and has good repeatability. Further investigations through the IR, DPV, AFM, UV–visible and fluorescence spectroscopies, SEM, EDX and TEM techniques confirmed the binding capabilities of the 3 with Cu(II).

Silica-supported indole derived fluorometric chemosensor for the selective detection of Ag+ ions in aqueous solution

A novel organic-inorganic hybrid nanomaterial-based fluorescent chemosensor was synthesized by functionalizing rice husk-derived nanosilica with 3-(aminopropyl)triethoxysilane (APTES) and then grafting it with para-aminobenzoic acid, benzaldehyde, and indole organic moieties, respectively, to detect Ag+ ions in the aqueous medium. Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), fluorescence spectroscopic techniques, and thermogravimetric analysis (TGA) were used to characterize the synthesized material. The synthesized ligand showed highly selective and sensitive fluorescence-enhanced, "Turn-ON" chemosensor activity towards Ag+ ions at an excitation wavelength of 355 nm, with observable variations in fluorescence intensity compared to other metal ions. The detection limit for Ag+ was 1.94 × 10−4 M, and the limit of quantification was calculated as 6.49 × 10−4 M. These findings suggest that the silica-supported indole derivative (Indole-APTES@nanosilica) can be utilized as a sensitive and selective, Turn-ON fluorometric chemosensor for detecting Ag+ ions.

Exploration of the Tungsten(VI) complex of hydroxamic acid's RNA binding affinity, histone deacetylase inhibitory activity, and antioxidant activity

Ribonucleic acid (RNA) is a versatile molecule, thus the opportunity to target RNA is abundant. Hence, the current exploration deals with the interaction of a metal complex of hydroxamic acid with t-RNA, along with HDAC8 inhibition activity and antioxidant activity. Binding characteristics of complex 1, Bis(N-phenylbenzohydroxamato)Tungsten(VI), which is represented as [N-PBHA-W(VI)] with Torula yeast RNA (t-RNA) were investigated by using several types of spectroscopic techniques, measurements of viscosity, and computational study through molecular docking. The intrinsic binding constant, Kb was evaluated for complex 1 which shows changes in the shift of absorption spectra, and the intensity of spectra also varies. From fluorescence measurement, the binding constant of complex 1 towards t-RNA is 6.64 ± 0.05 × 105 M−1. Stern-Volmer quenching constant was calculated. Two displacement method sare performed by using the fluorescence spectroscopic technique which discloses a groove mode of binding. The groove mode of binding is also uncovered by viscosity measurement, which was observed by increasing the complex 1 concentration. An electrochemical investigation was carried out by using cyclic voltammetry. A circular dichroism study was also employed, which further suggests a strong binding occurred between the metal complex used and t-RNA. The docked postures of t-RNA with complex 1 expose the strong interactions which suggest a minor groove mode of binding. All the experimental evidence indicates that the interaction of complex 1 with t-RNA is a minor groove mode of binding which complements the molecular docking results. In-silico studies of HDAC8 inhibition activity of complex 1 reveal that the inhibition takes place mainly by hydrophobic interaction. DPPH–radical scavenging method employed for the antioxidant activity.

Appling fluorescence spectroscopy with absolute principal component coefficient to explore dynamic migration of DOM fractions from an urbanized river during torrential rainfall

This research delves into the dynamic interplay between urbanization and the characteristics of Dissolved Organic Matter (DOM) in the Anyang River, particularly under the stress of torrential rain. The motivation stems from a critical need to decipher how urban landscapes influence water quality, focusing on the intricate transformations and movements of DOM. Employing advanced fluorescence spectroscopy techniques like Excitation-Emission Matrices (EEM) and Parallel Factor Analysis (PARAFAC), the study meticulously differentiates DOM compositions in urban and agricultural settings. It unveils a pronounced distinction, with urban streams showing elevated proteinaceous DOM from wastewater, contrasting with the humic substances prevalent in agricultural runoff. The analysis also captures how intense rainfall events catalyze significant shifts in DOM profiles, thereby emphasizing the need for tailored water quality management strategies in urbanized catchments. This comprehensive approach not only bridges gaps in understanding the urban impact on riverine ecosystems but also sets a foundation for future research and policy development in the face of escalating environmental changes.

DFT, real sample and smartphone studies of fluorescent probe, N-doped carbon quantum dots for sensitive and selective detection of bilirubin

In this investigation, nitrogen-doped carbon quantum dots (NCQDs) were successfully synthesized, showcasing robust fluorescence characteristics and photostability. These NCQDs demonstrated biocompatibility and were utilized as effective sensing agents for bilirubin (BR) detection. Their remarkable Stern-Volmer constant, attributed to exceptional molar absorptivity and photoluminescence quantum yield, rendered them highly proficient in BR sensing. Various analytical techniques such as X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDS), transmission electron microscopy (TEM), scanning electron microscopy (SEM), atomic force microscopy (AFM), and X-ray photoelectron spectroscopy (XPS) were employed to characterize the synthesized NCQDs, providing comprehensive insights into their structural, elemental, chemical composition, particle size, morphology, and optical properties. For sensing purposes, UV–Vis, steady-state, and time-resolved fluorescence spectroscopic techniques were utilized. The average particle size of the NCQDs was determined to be approximately 4.5 ± 0.3 nm. Notably, the Stern-Volmer constant (Ksv) for BR sensing from these quantum dots surpassed previously reported values in the literature. A linear relationship between emission intensity ratio and BR concentration within a range of 1.35–14.20 μM was observed. Accurate quantification of BR was achieved with a high Stern-Volmer constant (Ksv) S11 of approximately 7.48 × 105 M-1 and a limit of detection (LOD) of 4.86 μM. Leveraging the NCQDs as fluorescent probes, a highly sensitive and selective BR detection method based on the Förster resonance energy transfer (FRET) mechanism was established, with emission occurring at 360 nm. Density functional theory (DFT) studies supported FRET as the primary mechanism for fluorescence quenching, indicating electron transfer from NCQDs to BR, as evidenced by the energies of the lowest unoccupied molecular orbitals (LUMOs) of both NCQDs and BR. Real sample analyses of BR levels in human serum yielded a recovery range of 94.8 % to 104.21 %, validating the practical applicability of the method. Furthermore, smartphone-based BR detection using NCQDs achieved an LOD of 11.74 µM.

Study transport of hesperidin based on the DPPC lipid model and the BSA transport model

Hesperidin (HE), a significant flavonoid polyphenolic compound present in citrus plants, exhibits diverse pharmacological effects. Considering the crucial involvement of biological membranes and transporter proteins in the transportation and biological processes of HE, it becomes essential to comprehend the potential mechanisms through which HE interacts with membranes and transporter proteins. In order to simulate the process of active molecule transport, a cell membrane model consisting of 1,2-dipalmitoyl-n-glycero-3-phosphatidylcholine (DPPC) and a transporter protein model of bovine serum albumin (BSA) were employed for investigation. The present study aimed to investigate the mechanism of action of hesperidin (HE) in DPPC and BSA using fluorescence quenching, Fourier transform infrared spectroscopy (FTIR), and differential scanning calorimetry (DSC). The localization and interaction of HE within liposomes were also elucidated. Furthermore, the binding of BSA and HE was analyzed through UV/Vis absorption spectroscopy, fluorescence spectroscopy, infrared spectroscopy, and computational biology techniques. Computational biology analysis revealed that the binding between HE and BSA primarily occurred via hydrogen bonding and hydrophobic interactions. This study aimed to investigate the role and mechanism of HE in the DPPC cell membrane model and the BSA transporter protein model, thereby offering novel insights into the action of HE in DPPC and BSA.

Pyridine appended pyrimidine bis hydrazone: Zn2+/ATP detection, bioimaging and functional properties of its dinuclear Zn(II) complex

A pyridine functionalized pyrimidine-based system, H2P was successfully synthesized, characterized, and evaluated for its remarkable selective characteristics towards Zn2+ and ATP ions. The chemical sensing capabilities of H2P were demonstrated through absorption, fluorescence, and NMR spectroscopic techniques. The probe exhibited outstanding sensitivity when interacting with the ions, demonstrating relatively strong association constants and impressively low detection limits. The comprehensive binding mechanism of H2P with respect to Zn2+ and ATP ions was investigated using a combination of analytical methods, including Job's plot, NMR spectroscopy, mass spectrometry, and density functional theory (DFT) experiments. The interesting sensing ability of H2P for Zn2+/ATP ions was harnessed for live cell bioimaging and other diverse on-site detection purposes, including paper strips, cotton swabs, and applications involving mung bean sprouts. Further, the fluorescent probe demonstrated its effectiveness in detecting Zn2+ and ATP within live cells, indicating its significant potential in the realm of biological imaging applications. Moreover, the molecular configuration of the zinc complex (H2P–Zn2Cl4), derived from H2P, was elucidated using X-ray crystallography. This complex exhibited intriguing multifunctional attributes, encompassing its capability for detecting picric acid and for reversible acid/base sensing responses. The enhanced conducting behavior of the complex as well as its resistance properties were investigated by performing I–V characteristics and electrochemical impedance spectroscopic (EIS) experiments respectively.

C3-Symmetric Indole-Based Truxenes: Design, Synthesis, and Photophysical Studies.

In recent years, truxenes and related polyaromatic hydrocarbons (PAHs) have engrossed ample interest of the scientific community because of their ease of synthesis, functionalizations, and use as building blocks for the synthesis of fullerene fragments, liquid crystals, larger polyarenes, and C3-tripod materials. In the present work, we have disclosed an ingenious method for the construction of various indolo-truxene hybrid molecules in good yields (52-90%), by means of the acid-catalyzed cotrimerization, Friedel-Crafts acylation, and Fischer indole synthesis, and fully characterized them through the standard spectroscopic techniques. The photophysical properties of the thus-prepared compounds have also been investigated using steady-state absorption and fluorescence and time-resolved fluorescence spectroscopy techniques. Moreover, the density functional theory (DFT) and time-dependent DFT (TD-DFT) calculations have been studied to correlate them with the measured photophysical properties of the synthesized indolo-truxene derivatives.

Chemical Vapor Generation Non-Dispersive Atomic Fluorescence Spectrometry Technique for the Determination of Arsenic, Antimony, Selenium, and Mercury in Geological Samples by One-Time Digestion

In this paper, a one-time digestion method for the determination of arsenic (As), antimony (Sb), selenium (Se), and mercury (Hg) in geological samples was established. The optimum digestion conditions were determined with aqua regia-hydrofluoric acid (HF)-H3BO3 as a complexing open system, and the contents of As, Sb, Se, and Hg in the same digestion solution were determined by chemical vapor generation non-dispersive atomic fluorescence spectrometry (CVG-AFS). For the determination of As, Sb, Se, and Hg, the method detection limits were 0.53 mg/kg, 0.047 mg/kg, 0.009 mg/kg, and 0.00047 mg/kg, respectively, Δ log C was -0.043–0.048, and the %RSD values were lower than 8.5%, performing with good accuracy and precision. This digestion method avoids the fractional processing of each element, simplifies the digestion steps, saves pretreatment time, and improves efficiency. The digestion method also showed wide applicability, high sensitivity, and low detection limits, and it can be used as a quick method for monitoring and controlling heavy metal pollution in large-scale geological samples.

Exploring the mechanism of interaction between TBG and halogenated thiophenols: Insights from fluorescence analysis and molecular simulation

Thyroxine-binding globulin (TBG) plays a vital role in regulating metabolism, growth, organ differentiation, and energy homeostasis, exerting significant effects in various key metabolic pathways. Halogenated thiophenols (HTPs) exhibit high toxicity and harmfulness to organisms, and numerous studies have demonstrated their thyroid-disrupting effects. To understand the mechanism of action of HTPs on TBG, a combination of competitive binding experiments, multiple fluorescence spectroscopy techniques, molecular docking, and molecular simulations was employed to investigate the binding mechanism and identify the binding site. The competition binding assay between HTPs and ANS confirmed the competition of HTPs with thyroid hormone T4 for the active site of TBG, resulting in changes in the TBG microenvironment upon the binding of HTPs to the active site. Key amino acid residues involved in the binding process of HTPs and TBG were further investigated through residue energy decomposition. The distribution of high-energy contributing residues was determined. Analysis of root-mean-square deviation (RMSD) demonstrated the stability of the HTPs-TBG complex. These findings confirm the toxic mechanism of HTPs in thyroid disruption, providing a fundamental reference for accurately assessing the ecological risk of pollutants and human health. Providing mechanistic insights into how HTPS causes thyroid diseases.

Unveiling angular sweep total fluorescence Spectroscopy:A novel multidimensional technique for analysis of complex multi-fluorophoric systems

Analyzing multi-fluorophoric real systems without pre-processing is challenging, often addressed with unconventional fluorescence techniques and chemometrics. In this context, we introduce a novel addition - ‘Angular Sweep Total Fluorescence Spectroscopy (ASTFS),' to the arsenal of multidimensional steady-state fluorescence spectroscopic techniques. ASTFS utilizes a series of variable angle synchronous fluorescence spectra, strategically covering the fluorescence region between the first and second-order Rayleigh scattering ridges. The plot features a minimal data matrix size, avoids Rayleigh scattering signals, and incurs no blind regions. The study delves into the instrumental configurations for spectral acquisition, highlights the enhanced spectral resolution due to the band-narrowing effect, and discusses other notable features of the ASTFS plot. Further, this technique is reported to be effective in analyzing analytes in complex systems with strong background fluorescence, such as milk. The antibiotic— Norfloxacin is quantified via minimal pre-processing in milk samples and yields excellent analytical figures of merit.

Single molecule technique unveils the role of electrostatic interactions in ssDNA-gp32 molecular complex stability.

The exploration of single-strand DNA-binding protein (SSB)-ssDNA interactions and their crucial roles in essential biological processes lagged behind other types of protein-nucleic acid interactions, such as protein-dsDNA and protein-RNA interactions. The ssDNA binding protein gene product 32 (gp32) of the T4 bacteriophage is a central integrating component of the replication complex that must continuously bind to and unbind from transiently exposed template strands during the DNA synthesis. To gain deeper insights into the electrostatic conditions influencing the stability of the ssDNA-gp32 molecular complex, like the salt concentration or some metal ions proven to specifically bind to gp32, we employed a method that performs rapid measurements of the DNA-protein stability using an α-Hemolysin (α-HL) protein nanopore. We indirectly probed the stability of a protein-nucleic acid complex by monitoring the dissociation process between the gp32 protein and the ssDNA molecular complex in single-molecular electrophysiology experiments, but also through fluorescence spectroscopy techniques. We have shown that the complex is more stable in 0.5 M KCl solution than in 2 M KCl solution and that the presence of Zn2+ ions further increases this stability for any salt used in the present study. This method can be applied to other nucleic acid-protein molecular complexes, as well as for an accurate determination of the drug-protein carrier stability.

Delineating the core and surface state heterogeneity of carbon dots during electron transfer.

Exploring the heterogeneity of carbon dots (C-Dots) is challenging because of the existence of complex structural diversity, and it is a demanding task for the development and designing of efficient C-Dots for various applications. Herein, we studied the role of the core state and surface state of C-Dots in heterogeneity via the successful investigation of the electron transfer (ET) process between different (blue, green, and red) emitting C-Dots and an electron acceptor methyl viologen (MV2+) using steady-state and time-resolved fluorescence and ultrafast transient absorption (TA) spectroscopic techniques. Selective excitation in the steady-state and time-resolved mode shows that the ET ability of the core state is higher than that of the surface state. Moreover, the kinetics of MV+˙ generation was probed using TA spectroscopy after the excitation of the core and surface state, where we observed that the surface state becomes less efficient due to the presence of an oxygen-containing functional group in the surface state, which acts as an electron scavenger. Moreover, the heterogeneity of the core and surface state was explored through the detection of the MV+˙ generation yield after the irradiation of UV and visible light (exciting the core and surface state). The result indicates that the graphitic nitrogen content in the core state and the oxygen-containing functional group in the surface state play an important role in the heterogeneity in the structure and the ET process. Our findings on the fundamental understanding of the heterogeneity of different emissive C-Dots will provide a new way of designing and developing a metal-free light-harvesting system.

Eco-friendly novel deconvoluted synchronous spectrofluorimetric approach for the determination of favipiravir, levodropropizine and moxifloxacin hydrochloride as an effective therapeutic combination for COVID-19; application in laboratory prepared mixtures and spiked human plasma

In this work, a green, fast, and simple synchronous spectrofluorimetric approach has been developed to simultaneously determine favipiravir, levodropropizine, and moxifloxacin hydrochloride as co-administered medications for COVID-19 treatment in pure form and spiked human plasma. The synchronous fluorescence spectroscopy technique to analyze the studied drugs at Δλ = 110 nm enabled the determination of levodropropizine at 360 nm. Then, applying Fourier Self-Deconvolution to each spectra to measure favipiravir and moxifloxacin hydrochloride at peak amplitudes of 431 nm and 479 nm, respectively, without any interference. Favipiravir, levodropropizine, and moxifloxacin hydrochloride could be sensitively determined using the described approach over concentration ranges of 20–300 ng/mL, 10–600 ng/mL, and 50–500 ng/mL, respectively. The method's validation was carried out effectively in accordance with guidelines recommended by the ICH. Finally, the Eco-scale and Green Analytical Procedure Index (GAPI) techniques have been used to evaluate the greenness of the proposed method.

Shedding light on the ageing of extra virgin olive oil: Probing the impact of temperature with fluorescence spectroscopy and machine learning techniques

This work systematically investigates the oxidation of extra virgin olive oil (EVOO) under accelerated storage conditions with UV absorption and total fluorescence spectroscopy. With the large amount of data collected, it proposes a method to monitor the oil's quality based on machine learning (ML) applied to highly-aggregated data.EVOO is a high-quality vegetable oil that has earned worldwide reputation for its numerous health benefits and excellent taste. Despite its outstanding quality, EVOO degrades over time due to oxidation, which can affect both its health qualities and flavour. Therefore, it is highly relevant to quantify the effects of oxidation on EVOO and develop methods to assess it that can be easily implemented under field conditions, rather than in specialized analytical laboratories.The ML approach indicates that the two excitation wavelengths (480 nm) and (300 nm) exhibit the maximum relative change in fluorescence intensity during the ageing for the majority of the oils, thus identifying the wavelengths which are more informative for quality prediction. Also, the paper proposes a method for the prediction of olive oil quality using highly-aggregated data. Such a method is of interest because it paves the way to the realization of a low-cost, portable device for in-field quality control.The following study demonstrates that fluorescence spectroscopy has the capability to monitor the effect of oxidation and assess the quality of EVOO, even when the data are highly aggregated. It shows that complex laboratory equipment is not necessary to exploit fluorescence spectroscopy using the proposed method and that cost-effective solutions, which can be used in-field by non-scientists, could provide an easily-accessible assessment of the quality of EVOO.

Insight into the effect of synthesis parameters on the properties of graphene quantum dots for theranostic application

Ultra-small size and tunable optical properties of graphene quantum dots (GQDs) make them potentially suitable for theranostic applications. This investigation focuses on the effect of process parameters for the synthesis of GQDs of desirable optical properties for biomedical applications. The effect of the raw materials (graphene oxide sheets, glucose, and starch), reaction temperature (170 to 200 °C), and reaction time (3 to 6 h) was investigated on GQDs prepared through hydrothermal treatment. The dispersion of GQDs was characterized using dynamic light scattering (DLS), UV–visible, fluorescence, Raman, and X-ray photon spectroscopic techniques. The effect of the solution temperature and pH on the fluorescence properties of GQDs was also studied. The small hydrodynamic diameter (8.57 ± 1.47 nm) was observed with GQDs prepared from glucose using hydrothermal treatment time of 4 h at 190 °C (GQDs-D). The GQDs-D showed blue fluorescence (λem= 460 nm) with a quantum yield (QY) 67.09 ± 1.25%. The fluorescence characteristic of the GQDs was unchanged up to 6 months during storage at room temperature. GQDs-D revealed their antioxidant nature with a DPPH scavenging activity of 15.08 ± 2.07%. Despite having relatively low DPPH scavenging activity, excellent hemocompatibility (0.23 ± 0.04%) and colloidal stability against bovine serum albumin (BSA) showed GQDs-D as a safe nanocarrier for theranostic application. This was further substantiated with circular dichroism (CD) spectroscopy, which exhibited a preserved α-helical structure of BSA by GQDs-D. High-resolution transmission electron microscopic image of GQDs-D showed an average particle size of 5.70 nm and homogeneous particle distribution between 2–8 nm. Conclusively, all the results revealed the potential of GQDs-D for theranostic application.

Smart and low-cost fluorometer for identifying breast cancer malignancy based on lipid droplets accumulation.

The most common cause of breast cancer-related death is tumor recurrence. To develop more effective treatments, the identification of cancer cell specific malignancy indicators is therefore critical. Lipid droplets are known as an emerging hallmark in aggressive breast tumors. A common technique that can be used for observing molecules in cancer microenvironment is fluorescence microscopy. We describe the design, development and applicability of a smart fluorometer to detect lipid droplet accumulation based on the emitted fluorescence signals from highly malignant (MDA-MB-231) and mildly malignant (MCF7) breast cancer cell lines, that are stained with BODIPY dye. This device uses a visible-range light source as an excitation source and a spectral sensor as the detector. A commercial imaging system was used to examine the fluorescent cancer cell lines before being validated in a preclinical setting with the developed prototype. The outcomes indicate that this low-cost fluorometer can effectively detect the alterations levels of lipid droplets and hence distinguish between "moderately malignant" and "highly malignant" cancer cells. In comparison to prior research that used fluorescence spectroscopy techniques to detect cancer biomarkers, this study revealed enhanced capability in classifying mildly and highly malignant cancer cell lines.

Exploring protein-protein interactions and oligomerization state of pulmonary surfactant protein C (SP-C) through FRET and fluorescence self-quenching.

Pulmonary surfactant (PS) is a lipid-protein complex that forms films reducing surface tension at the alveolar air-liquid interface. Surfactant protein C (SP-C) plays a key role in rearranging the lipids at the PS surface layers during breathing. The N-terminal segment of SP-C, a lipopeptide of 35 amino acids, contains two palmitoylated cysteines, which affect the stability and structure of the molecule. The C-terminal region comprises a transmembrane α-helix that contains a ALLMG motif, supposedly analogous to a well-studied dimerization motif in glycophorin A. Previous studies have demonstrated the potential interaction between SP-C molecules using approaches such as Bimolecular Complementation assays or computational simulations. In this work, the oligomerization state of SP-C in membrane systems has been studied using fluorescence spectroscopy techniques. We have performed self-quenching and FRET assays to analyze dimerization of native palmitoylated SP-C and a non-palmitoylated recombinant version of SP-C (rSP-C) using fluorescently labeled versions of either protein reconstituted in different lipid systems mimicking pulmonary surfactant environments. Our results reveal that doubly palmitoylated native SP-C remains primarily monomeric. In contrast, non-palmitoylated recombinant SP-C exhibits dimerization, potentiated at high concentrations, especially in membranes with lipid phase separation. Therefore, palmitoylation could play a crucial role in stabilizing the monomeric α-helical conformation of SP-C. Depalmitoylation, high protein densities as a consequence of membrane compartmentalization, and other factors may all lead to the formation of protein dimers and higher-order oligomers, which could have functional implications under certain pathological conditions and contribute to membrane transformations associated with surfactant metabolism and alveolar homeostasis.

Anionic Hyperbranched Amphiphilic Polyelectrolytes as Nanocarriers for Antimicrobial Proteins and Peptides.

This manuscript presents the synthesis of hyperbranched amphiphilic poly (lauryl methacrylate-co-tert-butyl methacrylate-co-methacrylic acid), H-P(LMA-co-tBMA-co-MAA) copolymers via reversible addition fragmentation chain transfer (RAFT) copolymerization of tBMA and LMA, and their post-polymerization modification to anionic amphiphilic polyelectrolytes. The focus is on investigating whether the combination of the hydrophobic characters of LMA and tBMA segments, as well as the polyelectrolyte and hydrophilic properties of MAA segments, both distributed within a unique hyperbranched polymer chain topology, would result in intriguing, branched copolymers with the potential to be applied in nanomedicine. Therefore, we studied the self-assembly behavior of these copolymers in aqueous media, as well as their ability to form complexes with cationic proteins, namely lysozyme (LYZ) and polymyxin (PMX). Various physicochemical characterization techniques, including size exclusion chromatography (SEC) and proton nuclear magnetic resonance (1H-NMR), verified the molecular characteristics of these well-defined copolymers, whereas light scattering and fluorescence spectroscopy techniques revealed promising nanoparticle (NP) self- and co-assembly properties of the copolymers in aqueous media.