I22-Small angle scattering & Diffraction
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Eleni
Axioti
,
Nana A.
Berfi
,
Philippa L.
Jacob
,
Klara M.
Saller
,
Georgia L.
Maitland
,
Anisha
Patel
,
Sri Nithya
Paruchuri
,
Paul D.
Topham
,
Matthew J.
Derry
,
Shreyasi
Chatterjee
,
Benoit
Couturaud
,
Luciano
Galantini
,
Iolanda
Francolini
,
Valentina
Cuzzucoli Crucitti
,
Veeren M.
Chauhan
,
Robert J.
Cavanagh
,
Vincenzo
Taresco
Diamond Proposal Number(s):
[38357]
Open Access
Abstract: Recent studies have highlighted the limitations of conventional high degrees of PEGylation in drug delivery systems, including immune recognition and reduced efficacy. Approaches such as poly(ethylene glycol) (PEG) isomerization and shortening of PEG chains have emerged as strategies to mitigate anti-PEG immune responses while preserving key physicochemical properties required for drug delivery. Inspired by these advancements, this study aims to enzymatically synthesize new hybrid polymers incorporating a limited fraction of PEG and biosourced polyols, such as glycerol and diglycerol, as the hydrophilic counterpart, minimizing the amount of PEG by 50% (compared to our previous work). These novel adipate-based tetrapolymers, generated using four different starting materials, outperformed previous systems, offering a tunable and sustainable design for nanomedicine. By strategically limiting the PEG fraction, we preserved the functional benefits of PEGylation, including stealth and amphiphilicity, while advancing toward greener chemistry. The resulting biodegradable PEGylated polyesters were formulated from film rehydration of solid dispersions and increased the water solubility of the model drug curcumin via direct encapsulation of the compound in polymeric nanoparticles. The best performing polymer variant consisted of diglycerol, 1,6-hexanediol, and PEG combined with divinyl adipate (PEGDGA-Hex 50%). Its drug interactions, colloidal stability, biodegradability, and biocompatibility, in both in vitro (Caco2, human intestinal epithelial cells MCF-7, human breast cancer cells, and MDA-MB-231 late-stage triple-negative breast cancer cells) and invertebrate in vivo models that align with 3R principles (Caenorhabditis elegans and Drosophila melanogaster), support its potential use in systemic drug delivery.
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Jun 2026
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B21-High Throughput SAXS
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Diamond Proposal Number(s):
[29895]
Open Access
Abstract: A lipopeptide is designed that contains an epitope from simian virus T-antigen (SV40T, PKKKRKV) conjugated to an N-terminal palmitoyl (C16-) moiety, with the aim to act as an effective cell-penetrating lipopeptide, with additional aggregation propensity conferred by the lipid chain. A combination of cryo-TEM and small-angle X-ray scattering (SAXS) is used to show that the lipopeptide forms micelles, but mixtures with DNA lead to formation of fractal cluster-like co-assemblies due to intercalation of the DNA and peptide. Spectroscopic studies using fluorescence and circular dichroism (along with fiber X-ray diffraction) show that the peptide interacts with DNA and inserts into the groove. Confocal microscopy along with flow cytometry confirms delivery of DNA into both HeLa and mouse embryonic stem cells (mESCs) in pluripotent state, and the system shows excellent cytocompatibility as confirmed by MTT assays. Our data indicate that the lipopeptide may outperform the DNA transfection agent lipofectamine in DNA delivery into these stem cells and it enables DNA delivery into the cytoplasm and nucleus.
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Mar 2026
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B21-High Throughput SAXS
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Zimeng
Wang
,
Bei
Cheng
,
Christine
Tkaczyk
,
Michael
Newton
,
Shuolin
Cui
,
Bohdan
Andreiuk
,
Sanjib
Saha
,
Sabrina
Khan
,
Charles
Chen
,
Bin
Yang
,
Adam
Gamson
,
Sarah
Siddiqui
,
George
Thom
,
Antonio
Digiandomenico
,
Liping
Zhou
Open Access
Abstract: The recent success of mRNA COVID-19 vaccines using lipid nanoparticles (LNPs) underscores the potential of this technology for delivering gene-encoded biologics in humans. Passive immunization by in vivo expression of monoclonal antibodies (mAbs) could provide advantages over traditional vaccines due to its immediate protection and broader applicability. The use of mRNA allows for rapid development and easily scalable production, while in vivo expression of biologics (IVEB) reduces the complexity and cost associated with traditional mAb manufacturing by leveraging the body as a bioreactor. However, the requirement of cold chain storage and transportation remains a critical challenge for the distribution of mRNA-LNP formulations. Lyophilization of the mRNA-LNP formulations to produce dry powder offers a potential solution to this issue. In this study, we have investigated several lyophilization conditions for mRNA-LNP and evaluated their efficacy both in vitro and in vivo. Further physical stability and structure characterization provided insights into the structure-efficacy relationship. We successfully identified the lyophilization condition using 5 mM Tris buffer at pH 8 with 10% sucrose, which retained LNP particle size within an acceptable range, preserved mRNA encapsulation efficiency, and maintained consistent level of in vivo expressed mAbs after being stored at room temperature for a month.
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Mar 2026
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B21-High Throughput SAXS
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Diamond Proposal Number(s):
[37575, 38954]
Open Access
Abstract: Semaglutide is a therapeutically important lipopeptide that comprises a lipidated peptide with a glucagon-like peptide-1 (GLP-1) sequence, and may be prone to aggregation. We show that semaglutide in low pH 2.4 solutions forms β-sheet fibrils, in contrast to the oligomeric and micellar structures formed at higher pH. Based on cryo-TEM images showing twisted fibrils and the modeling of SAXS data (and with knowledge from fiber XRD) and molecular dynamics simulations, a model for the β-sheet structure is proposed, which comprises curved β-strands arranged in an antiparallel fashion around a core that comprises the lipidated lysine residue. This structure results from the patterning of the charged, polar, hydrophobic, and lipidated residues. Remarkably, it is possible to form a glass from the base form of semaglutide with crotonic acid, an organic salt capable of hydrogen bonding. Semaglutide glasses may have applications in biomedicine or therapeutics (for example, as slow-release depots).
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Mar 2026
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B21-High Throughput SAXS
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Diamond Proposal Number(s):
[29895]
Abstract: A hydrogel formed by a short peptide is presented that exhibits remarkable stimuli-responsiveness and plasticity, undergoing a morphological transformation from nanofibers to nanospheres in the presence of monovalent (Li+, Na+, K+) or trivalent (Al3+, Fe3+) metal ions under physiological conditions. The nanofibrillar structure of the hydrogel was examined using transmission electron microscopy (TEM), atomic force microscopy (AFM), small-angle X-ray scattering (SAXS), and X-ray diffraction (XRD) studies and atomistic molecular dynamics simulations, in complement, to explain the nanostructural transitions at the microscopic level. Interestingly, exposure to divalent metal ions (Mg2+, Ca2+, Co2+, Ni2+) induces a unique shrinking (syneresis) behavior, accompanied by a morphological shift to nanoribbons. Both simulations and SAXS analysis confirm that these ions cause a contraction in the packing of gelator peptides, significantly reducing the interpeptide distance. This ion-specific adaptability confers tunable physicochemical properties and morphological plasticity. Hydrogels incorporating mono- or trivalent ions exhibit enhanced thermal stability and mechanical strength relative to ion-free counterparts, underscoring the reinforcing role of metal coordination. Strikingly, shrunken gels formed in the presence of divalent ions display even greater stiffness than freshly prepared gels in the absence of any metal ions, suggesting that syneresis acts as a postassembly strengthening mechanism. These findings highlight a versatile, stimuli-responsive soft material in which ion-peptide interactions orchestrate nanoscale morphology, mesoscale network architecture, and macroscopic mechanical performance-opening avenues for adaptive hydrogel systems in targeted biomedical, sensing, and controlled-release applications.
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Feb 2026
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B21-High Throughput SAXS
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Open Access
Abstract: Amphiphilic compounds are important in many fields including pharmaceutical processes and development. Synthetic surfactants are often toxic to biological systems and frequently display poor biodegradability. Biosurfactants, such as lipopeptides and bile salts, on the other hand, can offer superior properties with regard to toxicity, biodegradability, and efficiency. Lipidated peptides are also gaining interest as therapeutic agents, as they can offer enhanced pharmacokinetic properties, compared with native peptides. The amphiphilic nature of lipidated peptides suggests that they may self-assemble into micellar structures, which can influence formulation stability and biological performance. Understanding the aggregation behavior of lipidated peptides is thus important for identifying and avoiding stability issues that could affect drug efficacy and safety. Structural characterization of self-assembled aggregates provides insight into aggregation mechanisms, which is valuable for identifying potential challenges during the production, storage, and administration of pharmaceutical peptides. By using small-angle X-ray scattering (SAXS), we have investigated the size and morphology of aggregates formed by MEDI7219, a bis-lipidated glucagon-like peptide-1 (GLP-1) analogue, in various aqueous solutions. We demonstrate that the lipopeptide MEDI7219 behaves as a surfactant with high spontaneous curvature that forms small micelles with a clear core-and-shell structure in aqueous solvents. The aggregation numbers of the micelles vary in the range of 5–8 and are found to be surprisingly insensitive to environmental conditions such as type of electrolyte and tonicity modifier, different buffers, and temperature, while exhibiting very low critical micelle concentrations (cmc).
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Feb 2026
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I22-Small angle scattering & Diffraction
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Diamond Proposal Number(s):
[33542]
Open Access
Abstract: Artificial cells assembled from materials such as hydrogels have emerged as platforms to replicate and understand biological functionalities, processes, and behaviors. However, hydrogels lack a lipid membrane, a vital property of cellular systems. Here we develop a process for the assembly of a fluid and stable lipid membrane which coats the hydrogel mesh network within the particle, through electostatically-mediated fusion of nanoscale lipid vesicles. This confers cell-mimetic and biotechnologically relevant properties upon microscale, cell sized, hydrogel artificial cells generated through microfluidics. We exploit the properties of the created membrane to augment existing hydrogel properties through permeability alteration and protection of the hydrogel from small molecule degraders. Furthermore, we show that the lipid membrane is compatible with organelle substructures within the hydrogels, which enables the exploitation of an enhanced material design space to build hydrogel artificial cells that increasingly mimic the organization of cells. This platform paves the way for producing next generation artificial cells and functional microdevices from interfaced hydrogel-lipid materials. Our technologies may underpin new opportunities for integrating membranes into hydrogel-based systems, inlcuding for drug delivery and tissue engineering.
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Jan 2026
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B21-High Throughput SAXS
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Open Access
Abstract: High-energy methods dominate the development of lipid nanoparticles but often require specialized equipment that increases production costs. Low-energy approaches, particularly those free of organic solvents, offer a promising alternative. This study aimed to obtain nanostructured lipid carriers (NLCs) using a solvent-free, low-energy process combining microemulsification and phase inversion. Cetearyl alcohol and PEG-40 hydrogenated castor oil were selected as the solid lipid and surfactant, respectively; the formulation and process were optimized through a Box–Behnken Design. Incorporation of the ionic surfactant extended colloidal stability, while the poloxamer in the aqueous phase enhanced steric stabilization. Resveratrol was efficiently encapsulated (E.E. = 98%), contributing to reduced particle size (291 nm), improved homogeneity (PDI = 0.25), and positive surface charge (+43 mV). Scale-up yielded stable particles carrying resveratrol with a mean size of 507 nm, PDI = 0.24, and ZP = +52 mV. The optimized formulation remained stable for 90 days at 8 °C. In vitro release demonstrated a sustained and controlled release profile, with significantly lower resveratrol release compared to the free compound. Thermal analysis confirmed drug incorporation within the lipid matrix, while transmission electron microscopy (TEM) revealed spherical particles (~200 nm) and SAXS indicated a nanostructure of ~50 nm. Overall, this study demonstrates that solvent-free, low-energy processing can produce stable and scalable NLC formulations, successfully encapsulating resveratrol with favorable physicochemical properties and controlled release behavior. These findings highlight a simple, cost-effective strategy for developing lipid-based nanocarriers with potential applications in drug delivery.
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Jan 2026
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Krios I-Titan Krios I at Diamond
Krios II-Titan Krios II at Diamond
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Diamond Proposal Number(s):
[31371]
Open Access
Abstract: Magnetotactic bacteria, such as Magnetospirillum gryphiswaldense MSR-1, naturally produce magnetosomes—intracellular magnetic nanoparticles that enable navigation within geomagnetic fields. Magnetosomes hold significant potential for biomedical and biotechnological applications; however, key aspects of their biomineralization remain poorly understood. This study investigates how oxidative stress, induced by hydrogen peroxide and iron, influences magnetosome formation and bacterial physiology under aerobic and microaerobic conditions. Single-cell advanced microscopy and high-throughput techniques revealed that microaerobic conditions supported robust magnetosome production and larger magnetite crystals while maintaining low oxidative stress levels. In contrast, aerobic conditions suppressed magnetosome formation, reduced intracellular iron content, and increased reactive oxygen species (ROS) levels. High extracellular iron enhanced the formation of longer magnetosome chains in microaerobic cultures without causing toxicity but reduced cell viability under aerobic conditions. Hydrogen peroxide exposure caused mild damage and a 25% viability drop in magnetosome-producing cells but led to severe damage and an 80% viability drop in non-magnetosome-producing cells, along with chain fragmentation and smaller magnetite crystals. These results suggest that magnetosome-producing cells exhibit greater resilience to oxidative stress, potentially due to ROS scavenging properties of magnetosomes, and highlight the intricate interplay between oxidative stress, iron regulation, and magnetosome biomineralization. Single-cell analysis revealed heterogeneity in physiological responses, further demonstrating the complexity of these processes. These findings underscore the importance of monitoring physiological changes during production processes to enhance the efficiency and robustness of magnetosome synthesis. The insights gained provide a foundation for improving bioprocesses for large-scale production of high-quality magnetosomes, advancing their applications in biomedicine and biotechnology.
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Dec 2025
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B21-High Throughput SAXS
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Diamond Proposal Number(s):
[31378]
Open Access
Abstract: Amphiphilic compounds, such as phospholipids or surface-active substances, are present in biological systems and can be part of pharmaceutical formulations. As a consequence, all pharmaceutically active ingredients will encounter amphiphilic compounds, either in the formulation or after administration. With the growing interest in peptide-based pharmaceuticals, there is a need to enhance the understanding of the interactions between peptides and amphiphilic compounds.
In this particular study, we have chosen to study mixtures of the comparatively small cyclical octapeptide lanreotide and the conventional anionic surfactant sodium dodecylsulfate (SDS). This was done by examining the self-assembly structures formed in lanreotide-SDS mixtures using light scattering and small-angle X-ray scattering (SAXS).
Above the critical micelle concentration (cmc) of SDS, the large excess of SDS could solubilize all lanreotide and form small micelles with lanreotide attached to the interface. Upon dilution to concentrations below the cmc of SDS, a suspension with dispersed solid nanoparticles is formed. The solid nanoparticles grow in size with decreasing concentration and, eventually, precipitate. The precipitated material is arranged in a liquid crystalline micellar phase, consisting of small close-packed SDS micelles with peptide adsorbed at the interface.
We were able to conclude that lanreotide does not form mixed micelles with SDS, indicating that it lacks the amphiphilic properties required to integrate fully with SDS behaving as a cosurfactant. In contrast, lanreotide attaches to the interface of SDS micelles, resembling the interactions of polymers, proteins, and nucleic acids with surfactants.
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Nov 2025
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