Search Results (35)

Antibody–drug conjugates (ADCs) comprise a monoclonal antibody covalently bound to a cytotoxic payload by a linker. ADCs minimize off-target effects on healthy tissues, leveraging the specificity of monoclonal antibodies to deliver cytotoxic drugs to the intended tumor site. ADCs can be prone to poor behavior, including aggregation and misfolding, leading to poor efficacy, impaired pharmacokinetics, and immunogenicity. It is advantageous to understand the developability and potential liabilities of a protein candidate prior to costly in vivo studies or clinical trials. This review summarizes biophysical and structural techniques used to characterize ADCs and introduces emerging techniques aimed at accurately assessing the developability of protein candidates. Stability is commonly assayed using techniques like differential scanning calorimetry (DSC), differential scanning fluorimetry (DSF), or spectroscopic probes such as circular dichroism and intrinsic fluorescence. Drug-to-antibody ratio (DAR) is a critical parameter that can be measured using absorbance spectroscopy or chromatographic analysis. Aggregation and self-association can be probed using scattering techniques such as dynamic light scattering (DLS), static light scattering (SLS), and size exclusion chromatography–multi-angle light scattering (SEC-MALS), as well as more specialized approaches such as fluorescence correlation spectroscopy (FCS) and analytical ultracentrifugation (AUC). Mass spectrometry (MS) provides extremely valuable insight into stability, covalent modifications, and, through approaches like hydrogen–deuterium exchange (HDX-MS), structural dynamics of ADCs. Looking forward, the use of biophysical assays in ex vivo matrices and strategic use of artificial intelligence/machine learning (AI/ML) approaches are likely to advance the efficient and rapid development of ADCs and other next-generation protein therapeutics. Full article

Starch systems and their extrudates can be used as edible films, carriers, and encapsulants for bioactive substances in various industries, primarily the food, medicine, and pharmacy industries. Using appropriate modification methods, it is possible to alter their physicochemical properties to improve specific functional parameters, thereby enhancing their application potential. The aim of this study was to characterize potato starch extrudates enriched with two types of edible oils (rapeseed or sunflower) at concentrations of 3%, 6%, and 9%. Chemical modification was carried out using K₂CO₃ as a catalyst. The structure of native and modified starch extrudates was examined using optical/confocal microscopy, FTIR, and LTNA (low-temperature nitrogen adsorption). Analogous starch dispersions were studied using static and dynamic light scattering, SLS/DLS, nephelometric methods, and electrophoretic mobility measurements to determine surface charge levels and stability. Additionally, viscosity curves were determined as a function of time and temperature. It was found that starch extrudates with 6% sunflower oil content showed optimal functional properties, characterized by greater stability, higher structural order, and better oil complexation. These findings directly translate into significant potential applications, including the development of functional products in the food industry. Full article

Studying nanoscale self-assembly in real time using external stimuli unlocks new opportunities for dynamic and adaptive materials. While electrostatic self-assembly is well-established, real-time monitoring of its structural evolution under light irradiation remains largely unexploited. In this study, we employ light-responsive azobenzene dyes (Acid Yellow 38, AY38) and pH-sensitive polyamidoamine (PAMAM) dendrimers to investigate the kinetics of electrostatic self-assembly under UV irradiation. Using a custom in situ small-angle neutron scattering (SANS) setup, we track the real-time morphological transformations of self-assembled structures with sub-minute resolution. We introduce two distinct pathways: method A (pre-irradiated cis-AY38 for controlled, slow kinetics) and method B (direct UV-induced self-assembly, fast kinetics). The results reveal that trans-cis isomerization kinetics dictate the rate of self-assembly, influencing aggregate stability, ζ-potential evolution, and final morphology. Structural analysis using dynamic and static light scattering (DLS and SLS) and SANS elucidates a transition from spherical to ellipsoidal morphologies governed by electrostatic and dipole-dipole interactions. These findings establish photoisomerization-driven self-assembly as a robust mechanism for tunable nanoscale architectures, paving the way for adaptive photonic materials, targeted drug delivery, and reconfigurable nanostructures. Full article

Τhe self-assembly behavior of a series of amphiphilic diblock copolymers, each consisting of a hydrophilic poly(N-vinyl pyrrolidone) (PNVP) block and a hydrophobic block derived from n-alkyl vinyl esters, namely poly(vinyl butyrate) (PVBu), poly(vinyl decanoate) (PVDc), and poly(vinyl stearate) (PVSt), in aqueous solutions was investigated. Dynamic and static light scattering (DLS and SLS) techniques were employed to monitor the micellization behavior. In addition, the self-assembled structures were observed with Transmission Electron Microscopy (TEM). The effect of the nature of the hydrophobic block, the copolymer composition and the copolymer molecular weight on the self-assembly properties was thoroughly examined. The encapsulation of curcumin and indomethacin within the dry cores of the micellar structures was conducted in aqueous solutions for all block copolymers at various curcumin/indomethacin-to-polymer mass ratios. UV-Vis spectroscopy was used to evaluate the drug-loading capacity and efficiency (%DLC and %DLE). In several cases, the encapsulation of both hydrophobic drugs was found to be nearly quantitative. Combined with the observed stability of the micellar structures, these findings suggest that the block copolymers demonstrate significant potential as carriers for drug delivery applications. Full article

The association behavior of amphiphilic block copolymers of N-vinyl pyrrolidone (NVP) and several vinyl esters (Ves) (PNVP-b-PVEs), as exemplified by vinyl butyrate (VBu), vinyl decanoate (VDc), and vinyl stearate (VSt), was studied in tetrahydrofuran (THF), which serves as the selective solvent for the PVE blocks. Static (SLS) and dynamic light scattering (DLS) techniques were adopted as the tools to investigate micellar properties and acquire information regarding the degree of association, the hydrodynamic radii, and the shape of the aggregates. In addition, CONTIN analysis provided insights concerning the association equilibria in THF solutions. The effect of the chemical structure of the corona-forming PVE block on the association process was investigated. Finally, the experimental results were compared with those obtained in previous studies describing the micellization properties of block copolymers consisting of PNVP and polymethacrylate blocks in the same selective solvent. Full article

Protein–polyelectrolyte nanostructures assembled via electrostatic interactions offer versatile applications in biomedicine, tissue engineering, and food science. However, several open questions remain regarding their intermolecular interactions and the influence of external conditions—such as temperature and pH—on their assembly, stability, and responsiveness. This study explores the formation and stability of networks between poly(acrylic acid) (PAA) and lysozyme (LYZ) at the nanoscale upon thermal treatment, using a combination of experimental and simulation measures. Experimental techniques of static and dynamic light scattering (SLS and DLS), Fourier transform infrared spectroscopy (FTIR), and circular dichroism (CD) are combined with all-atom molecular dynamics simulations. Model systems consisting of multiple PAA and LYZ molecules explore collective assembly and complexation in aqueous solution. Experimental results indicate that electrostatic complexation occurs between PAA and LYZ at pH values below LYZ’s isoelectric point. This leads to the formation of nanoparticles (NPs) with radii ranging from 100 to 200 nm, most pronounced at a PAA/LYZ mass ratio of 0.1. These complexes disassemble at pH 12, where both LYZ and PAA are negatively charged. However, when complexes are thermally treated (TT), they remain stable, which is consistent with earlier findings. Atomistic simulations demonstrate that thermal treatment induces partially reversible structural changes, revealing key microscopic features involved in the stabilization of the formed network. Although electrostatic interactions dominate under all pH and temperature conditions, thermally induced conformational changes reorganize the binding pattern, resulting in an increased number of contacts between LYZ and PAA upon thermal treatment. The altered hydration associated with conformational rearrangements emerges as a key contributor to the stability of the thermally treated complexes, particularly under conditions of strong electrostatic repulsion at pH 12. Moreover, enhanced polymer chain associations within the network are observed, which play a crucial role in complex stabilization. These insights contribute to the rational design of protein–polyelectrolyte materials, revealing the origins of association under thermally induced structural rearrangements. Full article

Accurate characterization of oleosome particle size distribution is needed for understanding their functionality in various applications. Traditionally, high-cost methods such as static laser diffraction and confocal or electron microscopy have been used. The current study presents a cost-effective alternative by combining optical microscopy (OM) with image analysis and dynamic light scattering (DLS) to evaluate particle size distribution in safflower (Carthamus tinctorius) oleosomes. Monodisperse and polydisperse standards (2 µm and 1–10 µm, respectively) were selected to validate instrument performance. The use of a smaller cuvette with a shorter path length in DLS extended its detection capabilities by minimizing multiple scattering and thermal effects. DLS and OM produced relatively consistent results, accurate particles’ diameters and distribution widths that agreed well with the standards. In contrast, static light scattering (SLS) showed strong sensitivity to the weighting method used (by number vs. by volume). In the case of polydisperse standard, volume-weighted SLS overestimated the particle size and yielded a broader distribution with a span of 2.2 compared to a span value of 0.8 as reported by the supplier. These findings highlight the importance of method selection and demonstrate the potential of combining DLS and OM as a practical and reliable approach for oleosome characterization. Full article

pH-responsive polyamidoamine (PAMAM) dendrimers are used as well-defined building blocks to design light-switchable nano-assemblies in solution. The complex interplay between the photoresponsive di-anionic azo dye Acid Yellow 38 (AY38) and the cationic PAMAM dendrimers of different generations is presented in this study. Electrostatic self-assembly involving secondary dipole–dipole interactions provides well-defined assemblies within a broad size range (10 nm–1 μm) with various shapes. The size and shape of these assemblies were determined using dynamic and static light scattering (DLS/SLS) and small-angle neutron scattering (SANS); ζ-potential measurements were performed to elucidate the charge characteristics, revealing the effective surface charge density of the nano-objects as an important parameter in the size and shape control. UV–vis spectroscopy and isothermal titration calorimetry (ITC) were employed to investigate the interaction on a molecular level and from a thermodynamic point of view. The results show that the amount of isomerized cis dye depends on the dendrimer generation because of a photoprotective effect through electrostatics for lower generations and through dipole–dipole interactions for higher generations; as the cis dye and trans dye bind with different strength, the amount of cis dye then again encodes the charge density and thereby the particle size and shape. Full article

The tetrazole moiety remains one of the most interesting scaffolds in the development of new high-energy density materials (HEDMs) because of its desired characteristics, such as high nitrogen content and heat of formation (HOF). The combination of several heterocycles with high HOF seems to be a promising strategy for obtaining energetic materials with superior properties. Herein, we report the synthesis and characterization of a tetrazole polymer, polymethylenetetrazole (PMT), as a potential HEDM. The compound was characterized using NMR, IR, and Raman spectroscopy. Its weight average molecular mass was obtained by static light scattering (SLS), and its physical properties by powder XRD analysis. The density, sensitivity to friction (FS), and impact (IS) of the compound were determined as well. The results of the thermal and energetic properties of PMT suggest that this polymer could be an insensitive explosive. Full article

In this work, the effect of sonication on the molecular characteristics of polyacrylic acid (Carbopol^® Ultrez 10), as well as on its rheological behavior in aqueous dispersions and microgels, was analyzed for the first time by rheometry, weight-average molecular weight (M_w) measurements via static light scattering (SLS), Fourier transform infrared (FTIR) spectroscopy and confocal microscopy. For this, the precursor dispersion and the microgels containing 0.25 wt.% of Ultrez 10 were sonicated in a commercial ultrasound bath at constant power and at different times. The main rheological properties of the microgel, namely, shear modulus, yield stress and viscosity, all decreased with increasing sonication time, while the microgel’s Herschel–Bulkley (H-B) behavior, without thixotropy, was preserved. Also, M_w of Ultrez 10 decreased up to almost one-third (109,212 g/mol) of its original value (300,860 g/mol) after 180 min of sonication. These results evidence a softening of the gel microstructure, which results from the reduction in the M_w of polyacrylic acid with sonication time. Separately, FTIR measurements show that sonication produces scission in the C-C links of the Carbopol^® backbone, which results in chains with the same chemistry but lower molecular weight. Finally, confocal microscopy observations revealed a diminution of the size of the microsponge domains and more free solvent with sonication time, which is reflected in a less compact and softer microstructure. The present results indicate that both the microstructure and the rheological behavior of Carbopol^® microgels, in particular, and complex fluids, in general, may be manipulated or tailored by systematic high-power ultrasonication. Full article

In this report, we present the results on the physicochemical characterization of cadmium telluride quantum dots (QDs) stabilized with glutathione and prepared by optimizing the synthesis conditions. An excellent control of emissions and the composition of the nanocrystal surface for its potential application in monoclonal antibody and biomarker testing was achieved. Two samples (QDYellow, QDOrange, corresponding to their emission colors) were analyzed by dynamic light scattering (DLS), and their hydrodynamic sizes were 6.7 nm and 19.4 nm, respectively. Optical characterization by UV-vis absorbance spectroscopy showed excitonic peaks at 517 nm and 554 nm. Photoluminescence spectroscopy indicated that the samples have a maximum intensity emission at 570 and 606 nm, respectively, within the visible range from yellow to orange. Infrared spectroscopy showed vibrational modes corresponding to the functional groups OH-C-H, C-N, C=C, C-O, C-OH, and COOH, which allows for the formation of functionalized QDs for the manufacture of biomarkers. In addition, the hydrodynamic radius, zeta potential, and approximate molecular weight were determined by dynamic light scattering (DLS), electrophoretic light scattering (ELS), and static light scattering (SLS) techniques. Size dispersion and the structure of nanoparticles was obtained by Transmission Electron Microscopy (TEM) and by X-ray diffraction. In the same way, we calculated the concentration of Cd²⁺ ions expressed in mg/L by using the Inductively Coupled Plasma Atomic Emission Spectrometry (ICP-OES). In addition to the characterization of the nanoparticles, the labeling of murine myeloid cells was carried out with both samples of quantum dots, where it was demonstrated that quantum dots can diffuse into these cells and connect mostly with the cell nucleus. Full article

Different microscopy and scattering methods used in the literature to determine the dimensions of cellulose nanocrystals derived from cotton and bacterial cellulose were compared to investigate potential bias and discrepancies. Atomic force microscopy (AFM), small-angle X-ray scattering (SAXS), depolarized dynamic light scattering (DDLS), and static light scattering (SLS) were compared. The lengths, widths, and heights of the particles and their respective distributions were determined by AFM. In agreement with previous work, the CNCs were found to have a ribbon-like shape, regardless of the source of cellulose or the surface functional groups. Tip broadening and agglomeration of the particles during deposition cause AFM-derived lateral dimensions to be systematically larger those obtained from SAXS measurements. The radius of gyration determined by SLS showed a good correlation with the dimensions obtained by AFM. The hydrodynamic lateral dimensions determined by DDLS were found to have the same magnitude as either the width or height obtained from the other techniques; however, the precision of DDLS was limited due to the mismatch between the cylindrical model and the actual shape of the CNCs, and to constraints in the fitting procedure. Therefore, the combination of AFM and SAXS, or microscopy and small-angle scattering, is recommended for the most accurate determination of CNC dimensions. Full article

High-solid-content polystyrene and polyvinyl acetate dispersions of polymer particles with a 50 nm to 500 nm mean particle diameter and 12–55% (w/w) solid content have been produced via emulsion polymerization and characterized regarding their optical and physical properties. Both systems have been analyzed with common particle-size-measuring techniques like dynamic light scattering (DLS) and static light scattering (SLS) and compared to inline particle size distribution (PSD) measurements via photon density wave (PDW) spectroscopy in undiluted samples. It is shown that particle size measurements of undiluted polystyrene dispersions are in good agreement between analysis methods. However, for polyvinyl acetate particles, size determination is challenging due to bound water in the produced polymer. For the first time, water-swelling factors were determined via an iterative approach of PDW spectroscopy error (Χ²) minimization. It is shown that water-swollen particles can be analyzed in high-solid-content solutions and their physical properties can be assumed to determine the refractive index, density, and volume fraction in dispersion. It was found that assumed water swelling improved the reduced scattering coefficient fit by PDW spectroscopy by up to ten times and particle size determination was refined and enabled. Particle size analysis of the water-swollen particles agreed well with offline-based state-of-the-art techniques. Full article

The detection and quantification of micro(nano)plastics in the marine environment are essential requirements to understand the full impacts of plastic pollution on the ecosystem and human health. Here, static light scattering (SLS) and dynamic (DLS) light scattering techniques are assessed for their capacity to detect colloidal particles with diameters between d = 0.1 and 0.8 µm at very low concentrations in seawater. The detection limit of the apparatus was determined using model monodisperse spherical polystyrene latex particles with diameters of 0.2 µm and 0.5 µm. It is shown that the concentration and size of colloids can be determined down to about 10⁻⁶ g/L. Light scattering measurements on seawater obtained from different locations in Western Europe show that colloidal particles were detected with DLS in seawater filtered through 0.8 µm pore size filters. The concentration of these particles was not higher than 1 µg/L, with an average diameter of about 0.6 µm. We stress that these particles are not necessarily plastic. No particles were detected after filtration through 0.45 µm pore size filters. Full article

Block copolymers of N-vinyl pyrrolidone (NVP) and benzyl methacrylate (BzMA), PNVP-b-PBzMA, were prepared by RAFT polymerization techniques and sequential addition of monomers. The copolymers were characterized by Size Exclusion Chromatography (SEC) and NMR spectroscopy. Differential Scanning Calorimetry (DSC), Thermogravimetric Analysis (TGA) and Differential Thermogravimetry (DTG) were employed to study the thermal properties of these copolymers. The micellization behavior in THF, which is a selective solvent for the PBzMA blocks, was examined. For comparison the self-assembly properties of the corresponding statistical copolymers, PNVP-stat-PBzMA, were studied. In addition, the association behavior in aqueous solutions was analyzed for the block copolymers, PNVP-b-PBzMA. In this case, the solvent is selective for the PNVP blocks. Dilute solution viscometry, static (SLS) and dynamic light scattering (DLS) were employed as the tools to investigate the micellar assemblies. The efficient encapsulation of the hydrophobic curcumin within the micellar core of the supramolecular structures in aqueous solutions was demonstrated by UV-Vis spectroscopy and DLS measurements. Full article