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In this study, ultrasound-assisted glycated ovalbumin (G-UOVA) based on natural deep eutectic solvents (NADES) was prepared using response surface optimization. The binding affinity of (-)-gallocatechin gallate (GCG) to native OVA (NOVA), ultrasound treated OVA (UOVA), glycated OVA (GOVA), and G-UOVA followed G-UOVA > GOVA > UOVA > NOVA. The effects of various modifications and GCG binding on the secondary structure, particle size, and thermal stability of NOVA were investigated. After ultrasound-assisted glycation, the emulsifying activity, emulsifying stability, foaming activity, and foaming stability of NOVA increased by 61.9 %, 119.0 %, 36.0 %, and 47.1 %, respectively, and further increased due to GCG binding. The encapsulation of native and modified OVA, particularly G-UOVA, significantly enhanced the antioxidant activity and bioaccessibility of GCG. The degradation of GCG encapsulated by G-UOVA decreased from 100 % to 20.9 % within 10 h. This study can lay the foundation for further applications of NADES in related fields., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Functional modification of drugs can significantly improve their efficacy and safety, thus enabling targeted therapy. Functional modifications based on polysaccharides can alter their molecular structure, and effectively enhance their functional properties and biological activities. Herein, we designed and synthesized cationic Laminarin (CLam) modified with polyethyleneimine (PEI) and explored its application as a vaccine adjuvant. The PEI modification resulted in a positively charged surface of CLam, which was mixed with model antigen (Ovalbumin, OVA) to form CLam/OVA nanoparticles with an optimal particle size of about 380.07 nm, a uniform distribution of the particle size and a stable system. In vitro experiments showed that the positive charge on the surface of CLam/OVA enabled it to be effectively internalized by bone marrow dendritic cells (BMDCs), promoted cell maturation, lysosomal escape, and the efficiency of antigen cross-presentation. Mechanically, CLam/OVA induces BMDC function via toll-like receptors, cytokine receptors, and chemokine-mediated signaling pathways. CLam/OVA induced stronger humoral and cellular immunity compared to the aluminum adjuvant. CLam/OVA induces higher levels of OVA-specific antibodies, generates cytotoxic T lymphocyte immune responses, and stimulates IFN-γ secretion. Overall, this study demonstrates that functionalization is critical for the rational design of polysaccharides to boost antigen-specific immune responses for more effective and long-lasting vaccination., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

An innovative integrated three-dimensional (3D) bioprinted gastric microtissue electrochemical biosensor was developed in this study for the detection of allergen ovalbumin (OVA). In this system, OVA triggers the release of histamine from gastric microtissue, which then undergoes a redox reaction on the electrode surface, leading to an increase in the peak current. Gelatin methacrylate hydrogel serves as a scaffold for the 3D culture of RBL-2H3 and PC-12 cells for partially restoring allergic reactions in the human body in vitro. Furthermore, gold nanoparticle-modified anodized aluminum oxide sieves macromolecular substances and facilitates sensor nano-analysis. Composites of cerium-based organometallic framework, MnO 2 , and gold nanoparticles significantly enhanced the sensitivity of the screen-printed carbon electrode. Under optimal experimental conditions, the detection limit for OVA was 0.042 μg/mL, with a linear range of 0.1-10.0 μg/mL. The fabricated sensor demonstrated high sensitivity, reliability, and simplicity, showcasing its broad potential for allergen detection applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Chicken breast products exhibit a rough texture and poor taste owing to the special fibrous tissue structure of chicken breast. Thus, adapting the taste of processed chicken breast products is pivotal to improving consumer satisfaction. Therefore, we used CaCl 2 and acyl-modified ovalbumin (AOVA) in a concerted effort to improve the gelation properties of chicken breast myofibrillar protein (MP). The investigation of molecular interactions and microstructure revealed the mechanism behind the changes in the physicochemical properties of the prepared gel. Molecular docking showed that AOVA enhanced the gel structure of MP through hydrogen bonding and salt bridges. The investigation results indicated that 0.30 mol/L CaCl 2 facilitated the unfolding of AOVA-MP molecules, increasing the number of disulphide bonds and hydrophobic interactions, as well as conferring better hardness, springiness, cohesiveness, and chewing properties to the AOVA-MP gel with a denser and more homogeneous gel network structure. This study provides new insights into the quality improvement in chicken breast mince products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Introduction: Tumor vaccines can activate tumor-specific immune responses to inhibit tumor growth, recurrence, and metastasis. However, the efficiency of antigen and adjuvants combined delivery to lymph nodes (LNs) is relatively low, leading to weak immune stimulation and tolerance. In this study, a tumor nanovaccine was constructed for the targeted dendritic cells (DCs)-mediated immunotherapy., Methods: Ovalbumin (OVA) antigen was first loaded into manganese-doped mesoporous silica nanoparticles (MMSNs) and coated with bacterial cytoplasmic membrane (BM), which was further inserted with mannose to prepare OVA@MMSNs@BM-Man nanovaccines. In vitro and in vivo experiments were conducted to assess their properties and function of the synthesized nanovaccines., Results: The nanovaccine can effectively target DCs in LNs by the combination of mannose with mannose receptor (CD205). BM serves as an immune adjuvant and co-delivers with OVA antigen, effectively improving antigen presentation efficiency. In an acidic environment, the Mn 2+ produced by the degradation of MMSNs can not only serve as an MR imaging agent but also activate the cGAS-STING pathway, followed by the release of IFN-β. The activated DCs further activate the body's cytotoxic T cells (CTLs), thereby exerting anti-tumor effects., The Conclusion: This study will provide a new idea for the construction of tumor nanovaccines., Competing Interests: The authors report no conflicts of interest in this work., (© 2025 Zhao et al.)

Proteins play crucial roles in nearly all biological activities, with their functional structures deriving from stable folded conformations. Protein denaturation, induced by chemical and physical agents, is a complex process where proteins lose their stable structures, thereby impairing their biological functions. Characterizing protein denaturation at the single-molecule level remains a significant challenge. In this study, we developed non-adhesive silicon nitride nanonets coated with polyethylene glycol to capture individual proteins. We utilized these nanonets to investigate the denaturation of ovalbumin induced by guanidine hydrochloride (Gdn-HCl) and lead chloride. The entire denaturation and renaturation processes of a single ovalbumin molecule were monitored via ionic current measurements through the nanonets. These non-sticky nanonets offer a versatile tool for real-time studies of structural changes during protein denaturation.

In recent decades, scientific methodologies applied in theCultural Heritage field have been growing, due to their pivotal role in guiding informed decisions concerning conservation strategies and daily maintenance. To achieve this goal, minimally/non-invasive quantitative and qualitative analyses are needed. However, the non-invasive and selective identification of proteinaceous binders and coatings in artworks represent an open issue in Cultural Heritage science. Herein, a novel miniaturized system is introduced, which consists of a label-free electrochemical immunosensor integrated with biocompatible Gellan gel. This method is intended to selectively and minimally invasively identify ovalbumin (OVA) on-site in paintings. The label-free immunosensor is made up on screen-printed electrodes (SPEs) by functionalizing the working electrode (WE) with a primary antibody (anti-ovalbumin) for the specific recognition of OVA. The presence of OVA produces antigen-antibody reaction, which results in the development of a bulky immunocomplex on the WE. This complex is quantified using square wave voltammetry (SWV) and a reversible redox probe: the current measured is inversely proportional to the OVA concentrations. The developed immunosensors showed good analytical performances when applied directly to painted mock-ups, exhibiting a limit of detection (LOD) of 1.6 ng mL -1 , a limit of quantification (LOQ) equal to 16 ng mL -1 , a working range between 0.01 and 0.4 μg mL -1 and selectivity for OVA over other protein components commonly present in painted artworks, including bovine serum albumin (BSA), collagen, and casein. The outcomes highlighted the dependability of the immunosensor in detecting OVA and the efficacy of Gellan gel as a streamlined method for extracting the target protein while preventing residue accumulation on the painting surface. This advancement suggests the potential of Gellan gel-coupled immunosensor systems as viable diagnostic alternatives for artwork management and preservation., Competing Interests: Declaration of competing interest X- The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

For understanding the behavior of the active substance in vivo, the near-infrared (NIR) spectral variations of ovalbumin (OVA) loaded in poly(N, N-dimethyl acrylamide) (PDMAA) hydrogel with temperature were investigated. Analyzing the spectra with improved resolution by continuous wavelet transform (CWT), the absorption variation of the peak at 4851 cm -1 arising from the α-helix of OVA with temperature was studied. The results show that a sharp decrease occurs at a lower temperature in PDMAA hydrogel, indicating that the unfolding of OVA in PDMAA hydrogel is facilitated. On the other hand, the intensity changes for the hydrogen-bonded water were consistent with that for the protein, providing evidence for facilitating the unfolding. Furthermore, the spectral feature of a new water structure with two hydrogen bonds was obtained from the spectra of OVA loaded hydrogel by independent component analysis (ICA). By analyzing the difference of the water structure with temperature and the side chain of hydrogels, it is demonstrated that the water structure may be a double hydration water surrounding both the protein and the methyl groups of the hydrogel. The easy dissociation of the double hydration water may be a crucial factor in facilitating the unfolding of proteins within hydrogels., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The effect of cold plasma (CP) treatment in promoting the covalent grafting of ovalbumin (OVA) with gallic acid (GA) were investigated, along with identifying the binding sites in the OVA-GA complex and exploring its potential for reducing the antigenicity of OVA. The results showed that the GA content of 22.97 ± 1.27 mg/g in OVA-GA complex was obtained following 60 s of CP treatment. Using LC-MS/MS, four regions (T 52 -R 59 , V 201 -K 207 , I 279 -R 285 , and V 281 -K 291 ) were identified, containing 12 GA binding sites in the OVA-GA complex. Additionally, a significant reduction in IgE binding capacity (70.83 ± 0.90 %) was observed, as confirmed by ELISA analysis. The masking/steric-hindrance effect on linear epitopes and the disruption of conformational epitopes of OVA as a result of GA grafting may be the key factors leading to the reduction in OVA antigenicity. This study highlights that promoting the grafting of polyphenols onto proteins using CP treatment is an effective strategy for reducing the antigenicity of protein allergens., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Dysphagia is a common functional disorder that limits the variety of available foods. This study explored the coordination assembly of tannic acid (TA) with Fe 3+ to form a metal-phenolic network (MPN) and developed ovalbumin (OVA)/MPN via a pH-driven method as a novel emulsifier to stabilize high internal phase Pickering emulsions (HIPPEs). Results indicated that, following pH-driven treatment, the OVA/MPN composite particles exhibited smaller sizes, enhanced electrostatic repulsion, and improved stability. UV-visible spectroscopy confirmed the successful assembly of MPN with OVA, while pH-driven processes facilitated MPN formation. Multi-spectral technology showed that MPN altered the intermolecular interactions and structural properties of OVA. The cooperatively modified OVA demonstrated superior interfacial wettability and emulsifying properties. Rheological studies revealed that all HIPPEs exhibited gel-like behavior and shear-thinning characteristics. HIPPEs stabilized by OVA, modified synergistically through pH-driven and MPN introduction, showed a dense network structure with higher viscosity, modulus, yield stress, and elasticity. IDDSI testing showed that HIPPEs with TA below 8 mg/mL had low-risk swallowing characteristics, while those with 12 mg/mL exhibited reduced rheological performance and failed the Level 4 dysphagia test. These findings provide crucial insights for the future development of HIPPEs suitable for individuals with dysphagia., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Storage of shell eggs converts natural ovalbumin (N-OVA) into its more thermostable forms (S-OVA). This conversion may be associated with deterioration in the foaming properties of the stored shell egg. Thus, the foaming behavior of N-OVA and S-OVA, especially their performance at different pH conditions, was conducted. Compared with N-OVA, S-OVA improved foaming ability by 29.04 % at pH 3.0 and exhibited rough foam. Regarding foaming stability, the conversion of N-OVA to S-OVA had a pronounced reduction effect, with foaming stability significantly decreasing by 28.48 %-100.00 % in pH 3.0-9.0. The spectroscopic analysis revealed that the alteration in the foaming properties of OVA was fundamentally attributed to its conformational change. Thermostable conformational transition provided S-OVA with smaller particle sizes, more flexible conformations, higher surface charge, and higher surface hydrophobicity. S-OVA at pH 3.0 showed a higher surface activity, indicating superior foaming ability. Moreover, N-OVA and S-OVA formed stiff and solid-like interfaces. Notably, N-OVA exhibited higher dilatational and elastic modulus, indicating a more compact and stable adsorption layer at air-water interface. Overall, thermostable conformational transition improved the interfacial activity of OVA and enhanced its foaming ability; however, overactive proteins were detrimental to the stabilization of its interfacial films., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: The structure of proanthocyanidins (PC) contains a large number of active phenolic hydroxyl groups, which makes it have strong antioxidant capacity. This study investigated the structural and functional properties of ovalbumin (OVA) modified by its interaction with PC., Results: It was found that on increasing the concentration ratio of PC to OVA from 10:1 to 40:1, the free amino and total sulfhydryl contents of OVA decreased from 470.59 ± 38.77 and 29.81 ± 0.31 nmol mg -1 to 96.61 ± 4.55 and 21.22 ± 0.78 nmol mg -1 , respectively, and the free sulfhydryl content increased from 7.65 ± 0.41 to 9.48 ± 0.58 nmol mg -1 . These results indicated that CN and CS bonds were formed and PC was covalently linked with OVA. The PC content in the OVA-PC conjugates increased from 281.93 ± 12.92 to 828.81 ± 46.09 nmol mg -1 on increasing the concentration ratio of PC to OVA from 10:1 to 40:1. The contents of α-helix and β-turn of OVA decreased, and the contents of β-sheet and random coil increased, confirmed by circular dichroism. The tertiary structure of OVA was also altered according to the results of fluorescence and ultraviolet absorption spectra. The surface hydrophobicity of OVA-PC conjugates decreased with increasing bound polyphenol content. The conjugation of OVA to PC significantly improved its emulsification and antioxidant activity and denaturation temperature., Conclusion: This study may provide valuable information for improving OVA's functional properties and its PC conjugates for applications in the food industry. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Vaccines are one of the greatest human achievements in public health, as they help prevent the spread of diseases, reduce illness and death rates, saving thousands of lives with few side effects. Traditional vaccine development is centered around using live attenuated or inactivated pathogens, which is expensive and has resulted in vaccine-associated illnesses. Advancements have led to the development of safer subunit vaccines, which contain recombinant proteins isolated from pathogens. Their short half-life and small size make most subunit vaccines less immunogenic. Here, we introduce a large pore 2D covalent organic framework (COF), PyCOFamide, as a promising solution for an effective subunit platform. Our study demonstrates that simple adsorption of a model antigen, ovalbumin (OVA), onto PyCOFamide (OVA@COF) significantly enhances humoral and cell-mediated immune response compared to free OVA. OVA@COF exhibited heightened immune cell activation and acts as an antigen reservoir, facilitating antigen-presenting cell trafficking to the draining lymph nodes, amplifying the humoral immune response. Additionally, the breakdown of the COF releases monomers that adjuvant the activation of immune cells vital to creating strong immunity. This platform offers a potential avenue for safer, more effective subunit vaccines., (© 2024 Wiley-VCH GmbH.)

Cancer vaccines utilizing nanoparticle (NP) structures that integrate antigens and adjuvants to enhance delivery and stimulate immune responses are emerging as a promising avenue in cancer immunotherapy. However, the development of cancer vaccines has been significantly hindered by the low immunogenicity of tumor antigens. To address this challenge, substantial efforts have been made in developing innovative adjuvants to elicit effective immune responses. In this study, we develop a NP cancer vaccine assisted by a polysaccharide derivative adjuvant, designed through a computational strategy, to evoke effective antigen-specific antitumor immunity. Using TLR4 as the putative receptor, we conducted a comprehensive evaluation of a prescreening library consisting of 34 inulin derivatives through docking and molecular dynamics simulation. Consequently, a new derivative, benzoylated inulin (InBz), is selected as the most promising TLR4 agonist. The adjuvant effect of InBz is evaluated by fabricating InBz NPs encapsulating the model antigen ovalbumin (OVA). In vitro, InBz-OVA NPs effectively activate the TLR4 signaling pathways and facilitate dendritic cell maturation, thereby enhancing the antigen delivery and presentation. In vivo, InBz-OVA NPs outperform a commercial aluminum-based adjuvant, elicit robust antibody titers, induce antigen-specific cytotoxic T lymphocytes, and achieve significant tumor suppression in murine models. Besides, the adjuvant effects of other representative derivatives, namely, acetylated and chloroacetylated inulin, with moderate and low potential from the library, are also chemically synthesized and experimentally evaluated and found to be in agreement with computational predictions, confirming the credibility of the strategy. This study provides an effective platform for the pursuit of efficient polysaccharide-based vaccine adjuvants.

To surmount the limitation of the instability of the currently reported water-in-water (W/W) emulsions, novel W/W emulsionss were constructed using amylopectin (AMP) and tara gum (TG) as the phases, and differently shaped ovalbumin (OVA) particles were used as stabilizers to improve the stability of W/W emulsions. Experiments displayed that the conformation of OVA could be changed by heating treatment, thus forming fibrous or spherical OVA particles that had the potential to stabilize TG-in-AMP (TG/AMP) emulsions. The emulsions had the best stability when the pH was 4 and the concentration of OVA particles was 3 %. Moreover, since ovalbumin fibril (OVAF) had better adsorption at the water-water interface than ovalbumin sphere (OVAS), OVAF-stabilized TG/AMP emulsion (OF-TE) had a relatively denser interfacial layer and exhibited more satisfactory ionic stability and physical stability than OVAS-stabilized TG/AMP emulsion (OS-TE). The rheological results demonstrated that OVAF and OVAS had little effect on the viscosity of TG/AMP emulsions. In brief, OVAF was more effective in improving the stability of TG/AMP emulsions than OVAS, and OF-TE did not show phase separation for at least 5 days. This study may be of great significance in improving the stability of food-grade W/W emulsions., Competing Interests: Declaration of competing interest The authors declare no conflict of interest and no competing financial interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A key challenge in vaccine development is to induce an effective and durable immune response. Live virus vaccines induce lifelong antibody responses; however, the immune responses induced by inactivated or subunit vaccines decrease gradually. Activation of the germinal center (GC) reaction, which generates long-lived plasma cells (LLPCs), is a key mediator of long-term antibody responses. To enhance the activation of GC, lymph node-targeted delivery of the vaccine is promoted by enhancing the deformability of the delivery vector. In this study, a double emulsion is designed with strong deformability and containing chitosan nanoparticles (CSNP) in the internal aqueous phase (W NP ) for efficient antigen loading, called W NP /O/W. The flexible oil layer and the internally loaded positively charged particles endow the emulsion with strong deformability, continuously enrich model antigen ovalbumin (OVA) in the lymph nodes, activate germinal center B (GC B) cells and T follicular helper (T FH ) cells, induce LLPCs, and obtain high-level antibody persistence for more than 5 months, which is significantly better than the traditional oil emulsion adjuvant. Concurrently, it also improves the immune-protective effect in aged mice. Altogether, these results indicate that W NP /O/W achieves lymph node targeted delivery by strengthening deformability, generating high-intensity antibody responses, and long-lasting immune protection., (© 2024 Wiley‐VCH GmbH.)

Nanoparticles are increasingly being used in the development of vaccines for disease prevention or treatment. Recent research has demonstrated that conjugating a protein onto the surface of nanoparticles can significantly increase its immunogenicity. Considering various pathogens that threaten human health, multivalent vaccines are often desirable. Up to now, nanoparticle-based vaccines are mostly limited to one protein per nanoparticle. No research has been conducted to explore the possibility of conjugating more than one protein onto the surface of a nanoparticle. Here we developed a specific conjugation strategy to conjugate multiple proteins to the PLGA/lipid hybrid nanoparticle surface. The maleimide-thiol Michael addition, Aizde-DBCO (Dibenzocyclooctyne), and TCO (trans-cycloctene)-Tetrazine click chemistry were employed to conjugate three different proteins, subunit keyhole limpet hemocyanin (sKLH), Ovalbumin (OVA), and cross-reactive material 197 (CRM 197 ), to the surface of PLGA/lipid hybrid nanoparticles (hNPs). The successful results of this study pave the way for developing multivalent vaccines against different pathogens., (© 2024 The Author(s). Journal of Biomedical Materials Research Part A published by Wiley Periodicals LLC.)

The development of cancer vaccines is at the forefront of cancer immunotherapy. Most existing strategies to induce an efficient anti-tumor immune response rely on molecular adjuvants and the incorporation of complex synthetic vectors into vaccine formulations. In contrast, this study introduces a one-step engineering technique to assemble the model antigen, Ovalbumin (OVA), into amyloid aggregates, leveraging biomimetic folding and aggregation to create non-fibrillar OVA globular aggregates and OVA amyloid-like fibrils as single-component, adjuvant-free vaccines. Notably, the OVA amyloid-like fibrils induced stronger immune responses compared to the native form, as evidenced by robust humoral immune reactions and the establishment of immune memory. These enhanced responses can be attributed to the self-adjuvant effect of the unique assembled structure, which preserves antigenic epitopes, improves antigen stability, facilitates antigen internalization, prolongs retention at the injection site, enhances antigen trafficking to the lymphoid organs, and promotes increased secretion of antibodies and cytokines. Furthermore, the efficacy of the vaccine was validated in a high OVA-expressing tumor model, demonstrating the potential of OVA amyloid-like fibrils as an effective vaccine for cancer immunoprevention. This minimalist self-adjuvant vaccine strategy holds promising implications for cancer immunotherapy and can inform the design of other protein antigen-based vaccines., (© 2024 Wiley‐VCH GmbH.)

Efficient delivery of therapeutic proteins remains a major challenge in developing effective immunotherapies and treatments for genetic disorders due to the limited tissue targeting capability of native proteins. In this study, the design and synthesis of protein nanocapsules (NCs) that achieve lung-specific delivery of therapeutic proteins are reported. These NCs are synthesized through a surface modification process that involves coating protein with functional monomers and cross-linkers, followed by in situ polymerization to create a protective shell on the protein surface with tailored surface chemistry. This approach preserves protein integrity and significantly enhances delivery efficiency and tissue specificity. Notably, it is shown that protein@NC with guanidine-rich surfaces exhibit exceptional lung-targeting capabilities. This is likely attributed to the formation of a vitronectin-rich protein corona, which facilitates receptor-mediated endocytosis by lung cells. The platform effectively delivers various proteins, such as ovalbumin, to antigen-presenting cells (APCs) in the lung, thereby enhancing antigen presentation and offering a promising strategy for cancer immunotherapy. These findings provide a significant advancement in tissue-specific protein delivery and hold the potential for targeted cancer immunotherapy., (© 2024 Wiley‐VCH GmbH.)

Food allergens are defined by their stability during digestion, with allergenicity largely influenced by resistance to enzymatic hydrolysis. Ovalbumin (OVA), a major egg protein, is a significant contributor to food allergies, particularly in children. Our previous work demonstrated that high hydrostatic pressure (HHP) treatment reduces OVA allergenicity by disrupting conformational epitopes and altering its structure. This study hypothesizes that HHP further influences OVA digestibility, allergenicity, and molecular structure during digestion. Results show that HHP treatment (600 MPa) reduced α-helix content by 16.1 % and increased β-sheet content by 38.4 %, enhancing free sulfhydryl groups and surface hydrophobicity. Hydrolysis and ELISA analyses confirmed that HHP accelerated enzymatic hydrolysis, significantly reducing OVA allergenicity. Molecular dynamics simulations revealed strengthened interactions between OVA and pepsin/trypsin, involving epitope residues. These findings indicate an association between HHP treatment and the modification of OVA's digestive stability and epitopes, suggesting its potential as a strategy for reducing allergenicity., Competing Interests: Declaration of competing interest The authors declare no competing financial interests that could have influenced the work presented in this paper., (Copyright © 2025 Elsevier Ltd. All rights reserved.)

During the preparation and development of selenium nanoparticles (SeNPs), natural bioactive peptides are added to enhance their physicochemical characteristics and functional properties. Among these properties, immunomodulatory activities, which include activating immune cells to strengthen immunity, constitute the major functions of the immune system. To obtain SeNPs with enhanced immunomodulation, ovalbumin peptide (OP) was used as a stabilizer, yielding OP-SeNPs. The physicochemical properties of OP-SeNPs were characterized. RAW264.7 macrophages were used as a model to investigate the immunomodulatory activity of OP-SeNPs. Results indicate that OP-SeNPs were zero-valent and amorphous, with a particle size of 82.23 ± 1.77 nm. SeNPs demonstrated positive interactions with the -OH, CO, CN, and NH groups of OP. In addition, OP-SeNPs activated RAW264.7 macrophages by increasing NO secretion and enhancing pinocytosis activity, indicating their ability to enhance immunomodulatory effects. Therefore, this study provides a theoretical basis for the construction and characterization of bioactive peptides and SeNP complexes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relations that influence the work reported in this paper., (Copyright © 2025. Published by Elsevier Ltd.)

Flavonoids are significant dietary components and have ability to coordinate with metal ions to produce novel drug discovery leads that are superior to those of the parent flavonoids. Here, in this report, we have synthesized chrysin-Cu(II) complex (as per reported article) and characterized it further with different analytical techniques. The synthesized complex was evaluated for radical scavenging and cell cytotoxicity studies where it exhibited enhanced activity as compared to bare chrysin. The interaction studies of the complex with ct-DNA ( K b ⁓ 10 5 M -1 ), human serum albumin (HSA) and ovalbumin ( K b ⁓ 10 4 M -1 ) were evaluated using multi-spectroscopic and molecular docking studies. Groove binding mode with ct-DNA was observed as confirmed from competitive displacement studies, viscosity measurement, melting temperature estimation and docking analyses. The complex exhibited comparatively higher affinity towards ct-DNA which indicated it efficient transportation by the carrier proteins and controlled release in the target DNA.

Food allergies affect millions of individuals worldwide, significantly impacting personal health and the economy. While avoiding allergenic foods remains the primary management strategy, consumers lack reliable means for immediate allergen detection in everyday dining settings. Here, we present iEAT2 (integrated Exogenous Allergen Test 2), an advanced electrochemical sensing system for rapid, on-site food allergen detection. Building upon our previous assay system, the iEAT2 features technical breakthroughs: i) a complete kit for sample processing, including a torsion device for food grinding, and ii) a new strategy for multi-electrode measurements, which enables the simultaneous detection of multiple allergens in a simplified electronic architecture. We designed a compact iEAT2 prototype capable of 16 electrochemical reactions. Experimental validation confirmed the independent electrochemical measurements in a simultaneous operation. Furthermore, the entire testing protocol was completed within 15 min, from allergen extraction to detection. The platform detected three common food allergens (gliadin, Ara h1, and ovalbumin) at concentrations below established allergic reaction thresholds. It also effectively identified cross-contamination events in real-world food samples. This technology may empower consumers to monitor food safety and improve its management., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier B.V. All rights reserved.)

Ovalbumin (OVA) is a high-risk allergen with complex tertiary structure in food samples. Here, we developed an accurate UPLC-MS/MS-based assay to improve OVA quantitative performance in processed foods. Full-length isotope-labeled OVA proteins (OVA-I) were synthesized using stable isotope labeling by amino acids in cell culture (SILAC) technique and employed as functional internal standards to ensure similar cleavage sites between internal standards and analytes. Subsequently, OVA and OVA-I fully expose their key cleavage sites by using 10 mol/L urea denaturation to correct errors caused by structural variations in different food matrices, the possible denaturation mechanism was also explored. Three quantitative peptides were selected for OVA detection, achieving 88 %-117 % recoveries with RSDs <9.36 % across three different spiking levels. The LOQs were set as 5 μg/g. OVA risk assessment in Chinese general population exhibited high exposure risks with intake level of most processed foods ranged from 0.35 to 215.66 mg., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Molecular weight (Mw) of ligand-mediated nanocarriers plays a pivotal role in their architecture and properties. In this study, self-assembled ovalbumin (OVA)-loaded nanoparticles were meticulously engineered by starch polyelectrolytes with different Mw. Results unveiled that, tailoring Mw of GRGDS pentapeptides-grafted carboxymethyl starch (G-CMS) displayed strong binding-affinity and transport efficiency through microfold cells (M cells) pathway in the simulated intestinal epithelial cell monolayer in which M cells were randomly located in the Caco-2 cells monolayer. Notably, nanoparticles assembled from G-CMS with relatively higher Mw exhibited more compact structures due to the stronger interactions between layers compared to that with relatively lower Mw, which rendered remarkably stable and only 19.01 % in vitro OVA leakage under conditions of the upper gastrointestinal tract. Subsequently, more intact nanoparticles reached M cells after in vitro digestion and exhibited higher transport efficiency through the M cells pathways (apparent permeability: 9.38 × 10 -5  cm/s) than Caco-2 cells, attributing to specific- and non-specific binding affinity towards M cells. Therefore, optimal Mw tailoring of starch polyelectrolytes can mediate the molecular interactions among their assembled layers and the interactions with M cells to balance the structural compactness, release and transport efficacy of nanoparticles, holding promise for advancing M cells-targeting oral delivery technologies., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In this study, ovalbumin (OV) and sodium alginate (SA), two macromolecular complexes, were coagulated into the emulsifier (OV/SA), which stabilized soybean oil by electrostatic interaction, hydrophobic interactions, and hydrogen bonding. The structure of OV/SA and properties of OV/SA Pickering emulsion were investigated. Additionally, the effect of emulsions on the gel and protein properties of hairtail surimi was studied. The results revealed that with the increasing concentration of OV/SA, the particle size and zeta potential value (negative value) of the emulsion initially decreased and then increased, while the rheological properties gradually improved. Compared with the surimi gel directly supplemented with soybean oil, the addition of emulsion enhanced gel strength, whiteness, water holding capacity, and hydrophobic interactions, resulting in a more stable gel network structure. In summary, incorporating emulsion into surimi at the same lipid content not only maintained its gel properties but also improved its color and compensated for lipid loss., Competing Interests: Declaration of competing interest The authors declared no conflicts of interest in connection with the research, authorship, and/or publication of this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Background: Simple, fast and low-cost paper-based analytical devices (PADs) have a good application prospect for point-of-care detection of GSH. However, effective immobilization of functional nanomaterials onto cellulose, as a critical factor in the construction of PADs, presents numerous difficulties and challenges., Results: In this study, we have developed an exceptionally straightforward and environmentally friendly synthetic approach by using ovalbumin (OVA) as a bio-mineralization template for the preparation of MnO 2 nanosheets. The MnO 2 nanosheets produced in the solution phase exhibited excellent intrinsic nano-enzyme activity and biodegradability. The OVA-MnO 2 nanosheets can effectively oxidize Amplex red in the absence of H 2 O 2 , enabling sensitive detection of GSH with a linear range of 5 nM-10 μM and a detection limit as low as 2.8 nM. Furthermore, we utilized this method to facilitate in situ synthesis of OVA-MnO 2 nanosheets directly on paper substrates. This approach eliminates the need for conventional stirring and centrifugation steps, greatly simplifying the fabrication process while reducing material usage and time expenditure. Characterization of the chemical composition and morphology confirmed the intimate growth of the 2D nano-enzymes on the cellulose fibers. Utilizing smartphone capabilities, the OVA-MnO 2 nanosheet-modified PAD enabled instrument-free detection of GSH, demonstrating high sensitivity (0.74 μM) and a wide linear response range (1-1000 μM)., Significance: The synthesis of MnO 2 nanosheets directly on cellulose substrates substantially streamlines the modification workflow of PADs and reduces detection costs, offering new avenues for clinical diagnostics of relevant diseases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Albumen, primarily composed of ovalbumin, is a vital, nutrient-rich ingredient in the food industry. Drying is a critical step in low-water-activity albumen powder production, allowing extended shelf-life and reduced costs in handling, transportation, and storage of albumen products. Traditional drying methods, such as spray drying (SD) and hot air drying (HAD), often degrade albumen. This study explores variable frequency contact ultrasonic drying (CUD) as a novel and green alternative, operating at a central frequency of 20 kHz with sound amplitudes of 0 %, 40 %, and 60 %, and temperatures of 40 °C and 60 °C. The drying kinetics, physical, and foaming properties of CUD-dried albumen proteins were compared with those of hot-air-, spray-, and freeze-dried (FD) samples. Compared to HAD, CUD significantly enhanced the drying process, as evidenced by a 240 % increase in effective moisture diffusivity, a 66-78 % reduction in activation energy (Ea), and a 27 % reduction in drying time. Moreover, CUD maintained higher protein integrity, evident from a 24-35 % decrease in enthalpies, more β-turn and random coil structures, and increased free sulfhydryl groups. Notably, CUD at 40 °C significantly improved foaming capacity by 88 %, and at 60 °C, it enhanced foaming stability by 34 %, outperforming other drying methods. Protein solubility of CUD-albumen was improved by 10-12 % compared to HAD and was slightly better than FD. CUD-albumen showed a brighter color with a 26 % lower browning index than the HAD samples. Overall, CUD emerges as an effective and sustainable method for drying high-protein materials, ensuring high-quality albumen powders., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

The development of nanovaccines capable of eliciting tumor-specific immune responses holds significant promise for tumor immunotherapy. However, many nanovaccine designs rely heavily on incorporating multiple adjuvants and carriers, increasing the biological hazards associated with these additional components. Here, this work introduces novel flexible nanocapsules (OVAnano) designed to mimic extracellular vesicles, primarily using the ovalbumin antigen and minimal polyethylenimine adjuvant components. These results show that the biomimetic flexible structure of OVAnano facilitates enhanced antigen uptake by dendritic cells (DCs), leading to efficient antigen and adjuvant release into the cytosol via endosomal escape, and ultimately, successful antigen cross-presentation by DCs. Furthermore, OVAnano modulates the intracellular nuclear factor kappa-B (NF-κB) signaling pathway, promoting DC maturation. The highly purified antigens in OVAnano demonstrate remarkable antigen-specific immunogenicity, triggering strong antitumor immune responses mediated by DCs. Therapeutic tumor vaccination studies have also shown that OVAnano administration effectively suppresses tumor growth in mice by inducing immune responses from CD8 + and CD4 + T cells targeting specific antigens, reducing immunosuppression by regulatory T cells, and boosting the populations of effector memory T cells. These findings underscore that the simple yet potent strategy of employing minimal flexible nanocapsules markedly enhances DC-mediated antitumor immunotherapy, offering promising avenues for future clinical applications., (© 2024 The Author(s). Advanced Healthcare Materials published by Wiley‐VCH GmbH.)

Antigen delivery via respiratory mucosal surfaces is an interesting needle-free option for vaccination. Nonetheless, it demands for the design of especially tailored formulations. Here, lipid/poly(lactic-co-glycolic) acid (PLGA) hybrid nanoparticles (hNPs) for the combined delivery of an antigen, ovalbumin (Ova), and an adjuvant, synthetic unmethylated cytosine-phosphate-guanine oligodeoxynucleotide (CpG) motifs, is developed. A panel of Ova/CpG-loaded lipid@PLGA hNPs with tunable size and surface is attained by exploiting two lipid moieties, 1,2 distearoil-sn-glycero-3-phosphoethanolamine-poly(ethylene glycol) (DSPE-PEG) and monophosphoryl lipid A (MPLA), with or without polyethyleneimine (PEI). It is gained insights on the lipid@PLGA hNPs through a combination of techniques to analytically determine the specific moiety on the surface, the spatial distribution of the components and the internal structure of the nanoplatforms. The collected results suggest that PEI plays a role of paramount importance not only in promoting in vitro antigen escape from lysosomes and enhancing antigen cross-presentation, but also in determining the arrangement of the moieties in the final architecture of the hNPs. Though multicomponent PEI-engineered lipid@PLGA hNPs turn out as a viable strategy for delivery of antigens and adjuvant to the respiratory mucosa, tunable nanoparticle features are achievable only through the optimal selection of the components and their relative amounts., (© 2024 The Author(s). Advanced Healthcare Materials published by Wiley‐VCH GmbH.)

Aluminum adjuvants remain the most commonly used vaccine adjuvants. Being rather effective in triggering humoral immunity, however, aluminum adjuvants usually show limited abilities in activating cellular immunities. Herein, by adding manganese ions during the preparation of aluminum adjuvant, a manganese-modified aluminum (Mn-Al) adjuvant is obtained, which can effectively stimulate both humoral and cellular immune responses. Such Mn-Al adjuvant can enhance antigen adsorption and promote antigen internalization by dendritic cells (DCs). Subsequently, the released Mn 2+ can activate the cyclic guanosine monophosphate-adenosine monophosphate synthase-stimulator of interferon genes pathway to further promote DC activation. When combines with the model antigen ovalbumin (OVA), the Mn-Al-adjuvantes vaccine can induce high levels of antigen-specific antibody titers and high proportions of antigen-specific cytotoxic T cells in vivo. Moreover, the Mn-Al-adjuvanted vaccine elicited stronger antigen-specific humoral and cellular immune responses than high-dose of the aluminum-based adjuvant. Additionally, immunization of mice with OVA in the presence of the Mn-Al adjuvant significantly inhibited the growth of B16-OVA tumors. Furthermore, when formulated with human papillomavirus antigens, Mn-Al-adjuvanted vaccines show better in vivo vaccination performance than aluminum-adjuvanted vaccines. Therefore, the manganese-modified aluminum adjuvant may thus become a new vaccine adjuvant with the potential to replace conventional aluminum adjuvants., (© 2024 Wiley‐VCH GmbH.)

In the past decade, cancer immunotherapy has revolutionized the field of oncology. Major immunotherapy approaches such as immune checkpoint inhibitors, cancer vaccines, adoptive cell therapy, cytokines, and immunomodulators have shown great promise in preclinical and clinical settings. Among them, immunomodulatory agents including cancer vaccines are particularly appealing; however, they face limitations, notably the absence of efficient and precise targeted delivery of immune-modulatory agents to specific immune cells and the potential for off-target toxicity. Nanomaterials can play a pivotal role in addressing targeting and other challenges in cancer immunotherapy. Dendritic mesoporous silica nanoparticles (DMSNs) can enhance the efficacy of cancer vaccines by enhancing the effective loading of immune modulatory agents owing to their tunable pore sizes. In this work, an emulsion-based method is optimized to customize the pore size of DMSNs and loaded DMSNs with ovalbumin (OVA) and cytosine-phosphate-guanine (CpG) oligodeoxynucleotides (CpG-OVA-DMSNs). The immunotherapeutic effect of DMSNs is achieved through controlled chemical release of OVA and CpG in antigen-presenting cells (APCs). The results demonstrated that CpG-OVA-DMSNs efficiently activated the immune response in APCs and reduced tumor growth in the murine B16-OVA tumor model., (© 2024 The Author(s). Small published by Wiley‐VCH GmbH.)

Objective: Microfluidics has emerged as a promising technique to prepare nanoparticles. However, the current microfluidic devices are mainly chip-based and are often integrated into expensive systems that lack on-the-spot versatility. The aim of this study was to set up a modular microfluidic system based on low-cost capillaries and reusable, easy-to-clean building blocks that can prepare poly(D,L-lactic-co-glycolic acid) (PLGA) nanoparticles with and without incorporated water-soluble biomacromolecules., Methods: A two-syringe system variant of the microfluidic system was set up to prepare PLGA particles and to investigate how the flow rates, solvents, and PLGA concentrations impacted the PLGA nanoparticle formation. A three-syringe system was designed to examine the incorporation of proteins into the PLGA particles., Results: The formation of the nanoparticles was affected by the PLGA concentration in the organic solvent, where an increasing concentration led to larger particle diameters (33-180 nm), and by the total flow rate, where an increase in the total flow rate led to smaller nanoparticles (197-77 nm). Using ultrapure water as the aqueous solvent resulted in precipitation at the outlet at higher PLGA concentrations. Aqueous poly(vinyl alcohol) created neutral particles in contrast to the negatively charged particles obtained with ultrapure water or an ethanol-water mixture. Incorporation of the proteins ovalbumin or lysozyme with a three-syringe system resulted in encapsulation efficiencies above 40%., Conclusion: A cheap and easily adjustable modular microfluidic system was developed to prepare PLGA nanoparticles with highly reproducible particle diameters that can effectively be loaded with proteins for drug and vaccine delivery., Competing Interests: Declarations. Conflict of Interest: The authors declare no conflicts of interest., (© 2024. The Author(s).)

Self-assembling protein nanoparticles are beneficial platforms for enhancing the often weak and short-lived immune responses elicited by subunit vaccines. Their benefits include multivalency, similar sizes as pathogens and control of antigen orientation. Previously, the design, preparation, and characterization of self-assembling protein vesicles presenting fluorescent proteins and enzymes on the outer vesicle surface have been reported. Here, a full-size model antigen protein, ovalbumin (OVA), was genetically fused to the recombinant vesicle building blocks and incorporated into protein vesicles via self-assembly. Characterization of OVA protein vesicles showed room temperature stability and tunable size. Immunization of mice with OVA protein vesicles induced strong antigen-specific humoral and cellular immune responses. This work demonstrates the potential of protein vesicles as a modular platform for delivering full-size antigen proteins that can be extended to pathogen antigens to induce antigen specific immune responses., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Julie A. Champion and Mariela R. Rodriguez-Otero report financial support was provided by National Science Foundation., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

With the continuous progress of nanotechnology in the field of tumor vaccines, immunotherapy has been regarded as one of the most powerful approaches for cancer treatment. Currently, DNA vaccines are used to efficiently deliver plasmids encoding tumor-associated antigens to antigen-presenting cells (APCs) and enhance the activation of immune cells. In this work, a series of aromatic sulfonyl small-molecule-modified polymers R-P based on low-molecular-weight polyethylenimine (PEI) were prepared, and their structure-activity relationship was studied. Among them, Ns-P with high transfection efficiency and low toxicity was applied to deliver antigen ovalbumin (OVA)-encoded plasmid DNA to APCs for triggering the immune activation of dendritic cells (DCs). It was also found that Ns-P could be used as an immune adjuvant to activate the STING pathway in DCs, integrating innate stimulating activity into the carrier to enhance antitumor immunity. Moreover, the modification of Ns-P/pOVA complexes with oxidized mannan could not only improve the biocompatibility of the complex, but also enhance the uptake of DCs, further inducing OVA antigen presentation and immune stimulation. In vivo antitumor assays indicated that Ns-P/pOVA/Man immunization could inhibit the growth of OVA-expressing E.G7 tumors in C57BL/6 mice. These results demonstrated that Ns-P/pOVA/Man is promising for gene delivery and immunotherapy application.

Spilled oil in ocean can spread to the shoreline and cause long-term impacts on the shoreline's ecological environment. Therefore, removing oil accumulated on shorelines is crucial. This study proposed an innovative ovalbumin (OVA) fluid-assisted method for the cleanup of oiled shoreline substrates. The oil removal efficiency of OVA fluids was systematically investigated. Higher concentrations of OVA fluids effectively enveloped and immobilized the oil, aiding in its separation from the sand surface. The increased temperature reduced the viscosity of emulsions, facilitating improved flow and oil removal. High salinity promoted the creation of oil particle aggregates molecules and facilitated the release of oil from the sand surface. The factorial analysis demonstrated that a high salt environment significantly enhances the combined impact of temperature and pH on oil removal performance. Different methods for the responsive separation of washing effluents were studied, and the most effective separation method was adjusting the pH of effluents to 4.54 (the isoelectric point of OVA). Separated precipitates exhibited good decomposition efficiency through thermal decomposition and biodegradation. OVA fluids boast advantages, such as low cost, easy recyclability, and non-toxicity, while ensuring high oil removal efficiency and making them a promising eco-friendly technique for the cleanup of oiled shorelines., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Modification of g-C 3 N 4 with metal-free biomaterials through an environmentally friendly, low-energy, facile, and rapid single-step method is desired for the preparation of photocatalysts with efficient activity and high selectivity of CO 2 reduction but remains a great challenge. Herein, we develop a phase-transitioned protein modification strategy for photocatalysts through superfast amyloid-like protein assembly on surfaces using a one-step sequential coating method. Metal-free carbon nitride/protein heterojunction composite photocatalysts (the phase-transitioned lysozyme (PTL), phase-transitioned bovine serum albumin (PTB), and phase-transitioned ovalbumin (PTO)-coated carbon nitride@SiO 2 (CN@SiO 2 ) and bioinspired carbon nitride hollow nanospheres (CN-HS) obtained by etching of CN@SiO 2 ) are prepared using lysozyme, bovine serum albumin, and ovalbumin. The insulator-semiconductor heterojunctions formed at the protein-carbon nitride interface promote the migration and separation of photogenerated charges. The exposed hydrophobic alkyl and aryl groups of the surface-modified protein enable the formation of a CO 2 -aqueous solution-photocatalyst three-phase interface on the catalyst surface and the exposed -NH 2 groups provide sites for CO 2 adsorption, which effectively increases CO 2 mass transfer and its adsorption as well as hydrophobicity, promoting CO 2 reduction and inhibiting hydrogen production. Therefore, protein modification effectively improves the CO 2 reduction activity and CO selectivity. For instance, compared to CN-HS, the CO yield of the PTL-modified CN-HS (1346.5 μmol g -1 ) increased by 24.5 times and the CO selectivity reached 90.5%. These findings represent a critical advancement in the surface modification of carbon nitride for CO 2 reduction and the design of bioinspired materials for various applications.

Cancer nanovaccines have emerged as an indispensable weapon for tumor treatment. However, insufficient immunogenicity and immunosuppression hamper the therapeutic effects of nanovaccines. Here, biodegradable nanovaccines (OMPP) composed of ovalbumin (OVA)-manganese oxide nanoparticles, amphiphilic poly(γ-glutamic acid) (γ-PGA), and ε-polylysine (PL) are constructed to realize enhanced cancer immunotherapy. Interestingly, amphiphilic γ-PGA and PL could serve as both carriers and immunoadjuvants to promote the cytosolic delivery of antigens and enhance the maturation of dendritic cells. Additionally, taking advantage of the photothermal property of OMPP, immunogenic cell death and in situ release of tumor-associated antigens can be triggered under near-infrared light irradiation for personalized tumor treatment. Moreover, OMPP nanovaccines can efficiently alleviate tumor hypoxia and downregulate programmed death-ligand 1 expression to reprogram the immunosuppressive tumor microenvironment. OMPP-mediated therapy has been shown to provoke robust immune responses to suppress B16-OVA melanoma and prevent postsurgical tumor recurrence. This work presents a facile strategy for the fabrication of nanovaccines by integrating carrier and adjuvant while exploring the inherent properties to promote antigen release and modulate immunosuppression, which demonstrates great potential for effective cancer immunotherapy.

Remarkable progress has been made in tumour immunotherapy in recent decades. However, the clinical outcomes of therapeutic interventions remain unpredictable, largely because of inefficient immune responses. To address this challenge and optimise immune stimulation, we present a novel administration route for enhancing the bioavailability of immunotherapeutic drugs. Our approach involves the development of an oral tumour vaccine utilising virus-like particles derived from the Hepatitis B virus core (HBc) antigen. The external surfaces of these particles are engineered to display the model tumour antigen OVA, whereas the interiors are loaded with cytosine phosphoguanosine oligodeoxynucleotide (CpG ODN), resulting in a construct called CpG@ OVA HBc with enhanced antigenicity and immune response. For oral delivery, CpG@ OVA HBc is encapsulated in a crosslinked dextran hydrogel called CpG@ OVA HBc@Dex. The external hydrogel shield safeguards the biomimetic virus particles from degradation by gastric acid and proteases. Upon exposure to intestinal flora, the hydrogel disintegrates, releasing CpG@ OVA HBc at the intestinal mucosal site. Owing to its virus-like structure, CpG@ OVA HBc exhibits enhanced adhesion to the mucosal surface, facilitating uptake by microfold cells (M cells) and subsequent transmission to antigen-presenting cells. The enzyme-triggered release of this oral hydrogel ensures the integrity of the tumour vaccine within the digestive tract, allowing targeted release and significantly improving bioavailability. Beyond its efficacy, this oral hydrogel vaccine streamlines drug administration, alleviates patient discomfort, and enhances treatment compliance without the need for specialised injection methods. Consequently, our approach expands the horizons of vaccine development in the field of oral drug administration., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The study aimed to assess the extent to which protein aggregation, and even the modality of aggregation, can affect gastric digestion, down to the nature of the hydrolyzed peptide bonds. By controlling pH and ionic strength during heating, linear or spherical ovalbumin (OVA) aggregates were prepared, then digested with pepsin. Statistical analysis characterized the peptide bonds specifically hydrolyzed versus those not hydrolyzed for a given condition, based on a detailed description of all these bonds. Aggregation limits pepsin access to buried regions of native OVA, but some cleavage sites specific to aggregates reflect specific hydrolysis pathways due to the denaturation-aggregation process. Cleavage sites specific to linear aggregates indicate greater denaturation compared to spherical aggregates, consistent with theoretical models of heat-induced aggregation of OVA. Thus, the peptides released during the gastric phase may vary depending on the aggregation modality. Precisely tuned aggregation may therefore allow subtle control of the digestion process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In this study, the enhancement of Pickering effect of ovalbumin (OVA) with bacterial cellulose nanofibers (BCNFs) prepared by electron beam irradiation was investigated and the environmental stability of oil-in-water Pickering emulsions stabilized by OVA/BCNFs complexes was explored by varying ratios of OVA/BCNFS (1:0.2, 1:0.4, 1:0.6, 1:0.8, 1:1) and oil phase concentrations (10 %, 20 %, 30 %, 40 %, 50 %, 60 %). Droplet sizes of Pickering emulsions were decreased with the increase of the proportion of BCNFs, while the viscosity and storage modulus (G') of Pickering emulsions were increased. The gel strength of Pickering emulsions was positively correlated with the oil phase content. Pickering emulsions stabilized by OVA/BCNFs complexes were endowed excellent environmental stability under varying pH, ionic strength, and thermal conditions. Moreover, after encapsulating curcumin in Pickering emulsions, the retention rates of curcumin were improved significantly during room temperature, UV light, and thermal treatment. The present study would contribute to the advancement of novel protein/polysaccharide stabilizers and offer novel insight for investigating the stability of Pickering emulsions and delivering lipophilic bioactive compounds., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Using high-resolution 3D printing, a novel class of microneedle array patches (MAPs) is introduced, called latticed MAPs (L-MAPs). Unlike most MAPs which are composed of either solid structures or hollow needles, L-MAPs incorporate tapered struts that form hollow cells capable of trapping liquid droplets. The lattice structures can also be coated with traditional viscous coating formulations, enabling both liquid- and solid-state cargo delivery, on a single patch. Here, a library of 43 L-MAP designs is generated and in-silico modeling is used to down-select optimal geometries for further characterization. Compared to traditionally molded and solid-coated MAPs, L-MAPs can load more cargo with fewer needles per patch, enhancing cargo loading and drug delivery capabilities. Further, L-MAP cargo release kinetics into the skin can be tuned based on formulation and needle geometry. In this work, the utility of L-MAPs as a platform is demonstrated for the delivery of small molecules, mRNA lipid nanoparticles, and solid-state ovalbumin protein. In addition, the production of programmable L-MAPs is demonstrated with tunable cargo release profiles, enabled by combining needle geometries on a single patch., (© 2024 The Author(s). Advanced Materials published by Wiley‐VCH GmbH.)

Purpose: Immunotherapy emerges as a promising frontier in cancer therapy and prevention. This study investigates the capacity of tumor-antigenic nanoparticles, specifically ovalbumin-tethered spiked virus-like poly(lactic-co-glycolic acid) nanoparticles (OVA-sVLNP), to effectively elicit humoral and cellular immune responses against tumors., Methods: OVA-sVLNP were synthesized through thiol-maleimide crosslinking using a single emulsion method. Comprehensive characterization was performed through Nuclear Magnetic Resonance (NMR), dynamic light scattering, Cryo-electron microscopy (Cryo-EM), confocal microscopy, and flow cytometry. Immunogenicity was evaluated using an enzyme-linked immunosorbent assay (ELISA) for quantifying immunoglobulin levels (IgG, IgG1, IgG2a) and cytokines in mouse sera. Flow cytometry profiled cellular immune responses in mouse spleens, and organ biosafety was assessed using immunohistochemistry and hematoxylin and eosin (H&E) staining., Results: OVA-sVLNP had a mean particle size of 193.8 ± 11.9 nm, polydispersity index of 0.307 ± 0.04, and zeta potential of -39.6 ± 10.16 mV, remaining stable for one month at 4°C. In vitro studies revealed significant upregulation of CD80/CD86 in dendritic cells, indicating robust activation. In vivo, the optimal concentration (V25) induced potent IgG, IgG1, and IgG2a antibodies, significant populations of CD3 + CD4 + , CD3 + CD8 + , and a rare subset of CD3 + CD4 + CD8 + memory T cells. Notably, Th9 induction resulted in the secretion of IL-9, IL-10, and other cytokines, which are crucial for orchestrating cytotoxic T cell activity and antitumor effects. Overall, higher doses did not improve outcomes, highlighting the significance of optimal dosing., Conclusion: This study demonstrated potent immunogenicity of OVA-sVLNP, characterized by the induction of specific IgG antibodies and the stimulation of cellular immune responses, particularly tumor-killing Th9 cells. The simplicity and cost-effectiveness of the manufacturing process augment the potential of OVA-sVLNP as a viable candidate for antitumor vaccines, opening new avenues for cancer prevention and cell-based therapeutic strategies., Competing Interests: The authors declare no conflicts of interest., (© 2024 Lin et al.)

In order to achieve a therapeutic effect, many drugs have to reach specific cellular compartments. Nanoscale drug delivery systems extend the circulation time, reduce adverse effects and thus improve tolerability compared to systemic administration. We have developed two types of albumin-coated nanocarriers equipped with built-in dyes to track their cellular uptake and intracellular enzymatic opening. Using the approved antiprotozoal drug and STAT3 inhibitor Atovaquone (Ato) as prototype for a hydrophobic small molecule, we show that Ato-loaded ovalbumin-coated nanocapsules (Ato-nCap) preferentially enter human myeloid cells. In contrast, Ato nanocrystals coated with human serum albumin (Ato-nCry) distribute their cargo in all different immune cell types, including T and B cells. By measuring the effect of Ato nanocarriers on induced STAT3 phosphorylation in IL-10-primed human dendritic cells and constitutive STAT3 phosphorylation in human melanoma cells, we demonstrate that the intracellular Ato release is particularly effective from Ato nanocrystals and less toxic than equal doses of free drug. These new nanocarriers thus represent effective systems for intracellular drug delivery.

Protein adsorption behavior was examined on poly( N -isopropylacrylamide-co-sodium methacrylate)-based hydrogels at different temperatures: 5, 20, and 37 °C, and pH: 4.5, 7, and 9.2. The hydrogels, whose covalent skeleton contains pendant anionic units due to the presence of the sodium methacrylate co-monomer, exhibited both thermo- and pH-sensitivity with different extents, which depended on the content of ionizable moieties and the cross-linker density. The hydrogel composition, temperature, and pH influenced the zeta potential of the hydrogels and their swelling properties. The proteins selected for the study, i.e., bovine serum albumin (BSA), ovalbumin (OVA), lysozyme (LYZ), and a monoclonal antibody (mAb2), differed in their aminoacidic composition and conformation, thus in isoelectric point, molecular weight, electrostatic charge, and hydrophobicity. Therefore, the response of their adsorption behavior to changes in the solution properties and the hydrogel composition was different. LYZ exhibited the strongest adsorption of all proteins with a maximum at pH 7 (189.5 mg ggel-1); adsorption of BSA and OVA reached maximum at pH 4.5 (24.4 and 23.5 mg ggel-1), whereas mAb2 was strongly adsorbed at 9.2 (21.7 mg ggel-1). This indicated the possibility of using the hydrogels for pH-mediated separation of proteins differing in charge under mild conditions in a water-rich environment of both the liquid solution and the adsorbed phase. The adsorption affinity of all proteins increased with temperature, which was attributed to the synergistic effects of attractive electrostatic and hydrophobic interactions. That effect was particularly marked for mAb2, for which the temperature change from 5 to 37 °C caused a twentyfold increase in adsorption. In all cases, the proteins could be released from the hydrogel surface by a reduction in temperature, an increase in pH, or a combination of both. This allows for the elimination of the use of salt solution as a desorbing agent, whose presence renders the recycling of buffering solutions difficult.

The inherent stochasticity associated with the hierarchical self-assembly of either native-like or partially-unfolded protein monomers leads to the formation of transient, morphologically-diverse prefibrillar species resulting in structurally-distinct polymorphic protein aggregates. High-resolution structural characterization of mature aggregates has revealed heterogeneous supramolecular packing of protofibrils within amyloid polymorphs. However, little is known about whether initial monomeric protein conformers engender polymorphism at the onset of aggregation. Here, we show that intrinsic conformational preference in aggregation-competent monomeric ovalbumin, an archetypal serpin, dictates fibrillar polymorphism by modulating aggregation pathways. Using fluorescence, FT-IR, and vibrational Raman spectroscopy coupled with dynamic light scattering and electron microscopy, we demonstrate that conformationally-diverse amyloidogenic monomers, formed via an interplay of electrostatic and hydrophobic interactions before the commencement of aggregation, play a crucial role in promoting amyloid polymorphism. Moreover, the monomeric conformational fingerprints, accrued at the onset of aggregation, persist and propagate during the formation of polymorphic amyloids. Our results delineate essential conformational characteristics of the monomeric protein preceding aggregation, which will have broad implications in the mechanistic understanding of amyloid strain diversity observed in disease-related proteins.

The utilization of cold plasma (CP) treatment to promote covalent conjugation of ovalbumin (OVA) and gallic acid (GA), as well as its functionality, were investigated. Results demonstrated that CP significantly enhanced the covalent grafting of OVA and GA. The maximum conjugation of GA, 24.33 ± 2.24 mg/g, was achieved following 45 s of CP treatment. Covalent conjugation between GA and OVA were confirmed through analyses of total sulfhydryl (-SH) group, Fourier transform infrared (FTIR) spectroscopy, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Unfolding of the OVA molecule occurred upon conjugation with GA, as evidenced by multiple spectroscopy analyses. Additionally, conjugation with GA resulted in significant improvements in the antioxidant activity and emulsifying properties of OVA. This study demonstrated that CP is a robust and sustainable technique for promoting the covalent conjugate of polyphenols and proteins, offering a novel approach to enhance the functional properties of proteins., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The presence of hierarchical suppressive pathways in the immune system combined with poor delivery efficiencies of adjuvants and antigens to antigen-presenting cells are major challenges in developing advanced vaccines. The present study reports a nanoadjuvant constructed using aluminosilicate nanoparticles (as particle templates), incorporating cytosine-phosphate-guanosine (CpG) oligonucleotides and small-interfering RNA (siRNA) to counteract immune suppression in antigen-presenting cells. Furthermore, the application of a metal-phenolic network (MPN) coating, which can endow the nanoparticles with protective and bioadhesive properties, is assessed with regard to the stability and immune function of the resulting nanoadjuvant in vitro and in vivo. Combining the adjuvanticity of aluminum and CpG with RNA interference and MPN coating results in a nanoadjuvant that exhibits greater accumulation in lymph nodes and elicits improved maturation of dendritic cells in comparison to a formulation without siRNA or MPN, and with no observable organ toxicity. The incorporation of a model antigen, ovalbumin, within the MPN coating demonstrates the capacity of MPNs to load functional biomolecules as well as the ability of the nanoadjuvant to trigger enhanced antigen-specific responses. The present template-assisted fabrication strategy for engineering nanoadjuvants holds promise in the design of delivery systems for disease prevention, as well as therapeutics., (© 2024 The Author(s). Small published by Wiley‐VCH GmbH.)

Pickering emulsions were co-stabilized by nanoliposome (NL) and thermally denatured ovalbumin (DOVA) based on the induction of OVA with strong particle characteristics through thermal denaturation. DOVA-NL particles were spherical and their sizes were mainly distributed between 50 and 100 nm. The surface tension and interfacial tension of DOVA-NL were significantly reduced, and the surface hydrophobicity, amphiphilicity and free -SH content of DOVA were enhanced after complexation with NL. The content of α-helix and β-sheet in DOVA decreased, whereas the content of β-turn and random coil increased after complexation with NL. Hydrophobic interactions, hydrogen bonding and electrostatic forces played a vital role in the interactions between NL and DOVA, leading to conformational changes in DOVA. The number of binding sites between NL and DOVA was more than one, and the interaction between NL and DOVA was exothermic and spontaneous. The emulsification index showed that DOVA-NL-stabilized Pickering emulsions (DNPE) were significantly more stable than DOVA-stabilized emulsions. DOVA-NL particles adsorbed at the oil-water interface and the droplet size of DNPE was smaller than that of DOVA-stabilized emulsions. This study suggests that it may be an effective strategy to improve the stability of Pickering emulsions through co-stabilization with NL and DOVA., Competing Interests: Declaration of competing interest We declare that we do not have any commercial or associative interest that represents a conflict of interest in connection with the work submitted., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Anthocyanins (AC) are vulnerable to degradation when affected by external factors. The present study employed ultrasound-assisted glycosylation of ovalbumin (OVA) and dextran (Dex) to generate conjugate carrier for AC to improve its stability. The results showed that sonication significantly improved the progression of Maillard reaction to OVA. Compared to traditional glycosylation, ultrasound treatment showed a higher degree of grafting, a lower number of free-SH, and smaller particle size and uniform distribution. The SDS-PAGE results indicated covalent interaction. Intrinsic fluorescence (INF), Fourier transform infrared spectroscopy (FTIR), and Circular dichroism (CD) analysis results suggested that ultrasound-assisted glycosylation altered the OVA structure. The scanning electron microscope (SEM) and X-ray diffractometer (XRD) observed that the ultrasound-assisted complex had a more compact and smoother structure and protein unfolding were better. The protein solubility increased significantly after glycosylation. Thermal gravimetric analysis (TGA) and Differential scanning calorimetry (DSC) indicated that the glycosylated conjugates can significantly improve the thermal stability of AC In addition, the AC showed an improved processing and storage stability when conjugated with glycosylated carrier. The glycosylated protein-anthocyanins complex may help provide new ideas and scientific basis for the development of naturally sourced anthocyanins-relevant products in pharmaceutical and food industry applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)