Protein Nanoparticles Research Articles

Ferritin-based disruptor nanoparticles: A novel strategy to enhance LDL cholesterol clearance via multivalent inhibition of PCSK9-LDL receptor interaction.

Hypercholesterolemia, characterized by elevated low-density lipoprotein (LDL) cholesterol levels, is a significant risk factor for cardiovascular disease. Proprotein convertase subtilisin/kexin type 9 (PCSK9) plays a crucial role in cholesterol metabolism by regulating LDL receptor degradation, making it a therapeutic target for mitigating hypercholesterolemia-associated risks. In this context, we aimed to engineer human H ferritin as a scaffold to present 24 copies of a PCSK9-targeting domain. The rationale behind this protein nanoparticle design was to disrupt the PCSK9-LDL receptor interaction, thereby attenuating the PCSK9-mediated impairment of LDL cholesterol clearance. The N-terminal sequence of human H ferritin was engineered to incorporate a 13-amino acid linear peptide (Pep2-8), which was previously identified as the smallest PCSK9 inhibitor. Exploiting the quaternary structure of ferritin, engineered nanoparticles were designed to display 24 copies of the targeting peptide on their surface, enabling a multivalent binding effect. Extensive biochemical characterization confirmed precise control over nanoparticle size and morphology, alongside robust PCSK9-binding affinity (KD in the high picomolar range). Subsequent efficacy assessments employing the HepG2 liver cell line demonstrated the ability of engineered ferritin's ability to disrupt PCSK9-LDL receptor interaction, thereby promoting LDL receptor recycling on cell surfaces and consequently enhancing LDL uptake. Our findings highlight the potential of ferritin-based platforms as versatile tools for targeting PCSK9 in the management of hypercholesterolemia. This study not only contributes to the advancement of ferritin-based therapeutics but also offers valuable insights into novel strategies for treating cardiovascular diseases.

Characterization of orange peel extract-cross linking whey protein nanoparticles and their influences on the physical and sensory properties of orange juice

This work explores the characterization of orange peel extract (OPE)-cross-linked whey-protein nanoparticles for enhancing the physical and sensory properties of orange juice. Three methods were employed to cross-link particle-forming peptides with citric acid, i.e., OPE charging–cross-linking–desolvation (A), cross-linking–OPE charging–desolvation (B), and OPE charging–desolvation–cross-linking (C). B method produced smaller particles (229 ± 11.53 nm) with higher encapsulation efficiency (67.16 ± 1.75%). Kinetic studies indicated predominantly diffusion-based release of OPE from the particles, B express the lowest diffusion rate with 10.20 ± 1.23% after 6 h incubation in NaCl. Using citric acid as a cross-linking agent enhanced the encapsulation efficiency. Moreover, functional orange juices were formulated using OPE and B capsules (500 mg/L). Under simulated in-vitro gastrointestinal conditions, the antioxidant activity was higher in B capsule supplemented juice comparing to OPE supplemented juice at oral, gastric, and intestinal phase which recorded of 69.43 ± 0.750, 85.66 ± 1.12 and 94.53 ± 2.00 mg ascorbic acid/100g respectively by DPPH and 80.91 ± 1.37, 132.01 ± 0.50 and 153.47 ± 2.63 mg ascorbic acid/100g respectively by ABTS. Sensory evaluation demonstrated the acceptability of OPE and cross-linked nanoparticles by the panelists. Cross-linking–OPE charging–desolvation provides high encapsulation efficiency with palatable physico-sensorial attributes in orange juice and offers a promising strategy for creating a nutritious beverage.

Preventing the impact of solute adsorption in Taylor dispersion analysis: Application to protein and lipid nanoparticle analysis

Taylor Dispersion Analysis (TDA) allows diffusion coefficient (D) or hydrodynamic radius (Rh) determination on a wide range of size between angstroms and about 300 nm. However, solute adsorption phenomena can affect the repeatability and reproducibility of TDA. Several numerical studies addressed the theoretical impact of solute adsorption in TDA, but very few experimental studies focus on this topic and no experimental methodologies were proposed so far to reduce the impact of adsorption in TDA. In this work, an experimental protocol, called plug-in-front TDA, consisting of adding the solute in the eluent at a lower concentration compared to the injected sample, was proposed to strongly limit the impact of adsorption on the Rh determination. This protocol was suggested based on the evidence that adsorption / desorption phenomena impacting narrow bore fused silica TDA in aqueous conditions are typically slow processes that can be counteracted by saturating the interaction sites during the experiments. Successful applications to proteins and mRNA lipid nanoparticles (LNP) in vaccine against Covid 19 and protein analysis were reported. TDA of proteins in conditions of strong interactions with the capillary surface was possible using the plug-in-front methodology. We anticipate that such experimental methodology will greatly help the experimentalist for implementing TDA in various applications.

A Single-Component Multilayered Self-Assembling Protein Nanoparticle Vaccine Based on Extracellular Domains of Matrix Protein 2 against Both Influenza A and B.

The development of an effective and broadly protective influenza vaccine against circulating and emerging strains remains elusive. In this study, we evaluated a potentially universal influenza vaccine based on single-component self-assembling protein nanoparticles (1c-SApNPs) presenting the conserved matrix protein 2 ectodomain (M2e) from influenza A and B viruses (IAV and IBV, respectively). We previously designed a tandem antigen comprising three IAV M2e domains of human, avian/swine, and human/swine origins (termed M2ex3). The M2ex3-presenting 1c-SApNPs conferred complete protection in mice against sequential lethal challenges with H1N1 and H3N2. To broaden this protection to cover IBVs, we designed a series of antigens incorporating different arrangements of three IAV M2e domains and three copies of IBV M2e. Tandem repeats of IAV and IBV (termed influenza A-B) M2e arrayed on the I3-01v9a 60-mer 1c-SApNP, when formulated with an oil-in-water emulsion adjuvant, generated greater M2e-specific immunogenicity and protective efficacy than the soluble influenza A-B M2e trimer, indicated by higher survival rates and reduced weight loss post-challenge. Importantly, one of the influenza A-B M2e SApNP constructs elicited 100% protection against a lethal influenza A/Puerto Rico/8/1934 (H1N1) challenge in mice and 70% protection against a lethal influenza B/Florida/4/2006 (Yamagata lineage) challenge, the latter of which has not been reported in the literature to date. Our study thus provides a promising M2e-based single-component universal vaccine candidate against the two major types of influenza virus circulating in humans.

Synthetic Protein Nanoparticles via Photoreactive Electrohydrodynamic Jetting.

Protein nanoparticles are an attractive class of materials for nanomedicine applications due to the intrinsic biocompatibility, biodegradability, and intrinsic functionality of their constituent proteins. Despite the clinical success of select protein nanoparticles, this class of nanocarriers remains understudied and underdeveloped compared to lipid and polymer nanoparticles due to challenges related to formulation optimization, large design space, and their structural complexity. In this work, a modular strategy for protein nanoparticle preparation based on the concept of photoreactive jetting is introduced. The process relies on continuous ultravioletirradiation during electrohydrodynamic (EHD) jetting of protein solutions that contain a homobifunctional photocrosslinker. Protein nanoparticles exhibit nanogel-like architectures comprised of proteins that are linked via synthetic moieties. Compared to conventional protein nanoparticles, this method reduces nanoparticle processing times to minutes, rather than hours to days. The inclusion of an emissive structural motif as the molecular scaffold of the photocrosslinker is used to study the supramolecular architecture of the stable nanoparticles via time-resolved fluorescence spectroscopy.

Fabrication and characterization of novel TGase-mediated glycosylated whey protein isolate nanoparticles for curcumin delivery

The aim of this study was to fabricate novel transglutaminase (TGase)-mediated glycosylated whey protein isolate (WPI) nanoparticles for the encapsulation and delivery of curcumin. The influences of glycosylation on the physiochemical properties, stability, bioavailability, and antioxidant properties of WPI nanoparticles loaded with curcumin were investigated. Composite nanoparticles exhibited uniform distribution and small particle sizes. The main driving forces for the formation of curcumin nanoparticles were electrostatic interactions, hydrogen bonding, and hydrophobic interactions. The encapsulation and loading efficiency of curcumin after TGase-type glycosylation were significantly increased in comparison to WPI-curcumin nanoparticles. Glycosylated WPI-curcumin nanoparticles had stronger antioxidant properties and stability to resist external environmental changes than WPI-curcumin nanoparticles. In addition, glycosylated WPI-curcumin nanoparticles showed a controlled release and enhanced curcumin bioavailability in vitro gastrointestinal digestion. This study provides novel insights for self-assembled glycosylated protein nanoparticles as delivery systems for protecting hydrophobic nutrients.

Antibacterial rice protein nanoparticles with a high curcumin loading for fruit preservation

Curcumin (Cur) exhibits potential as a food preservative because of its natural antibacterial and antioxidant properties. Nevertheless, its practical application is challenging due to poor water solubility and physicochemical instability. In this study, we prepared Cur-loaded protein nanoparticles (RCs) with a high loading capacity. The water-dispersible nanoparticles (10–30 nm) enclosing Cur with a loading capacity of up to 302 mg/g protein were formed by neutralizing rice proteins (RPs) from alkali-unfolded RPs utilizing cation exchange resin in the presence of Cur. Compared to free Cur, RCs improved the heat-stability and photo-stability of Cur by 26.7% and 23.2%, respectively. The RCs demonstrated significantly higher antibacterial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) compared to free Cur, with inhibition rates exceeding 94.45% and 91.00%, respectively. Additionally, RCs exhibited superior antioxidant activities across multiple assays, with RCs5 showing increases of 122.22% (DPPH), 64.99% (ABTS), and 94.72% (FRAP) compared to free curcumin. RCs significantly preserved freshly-cut apples, with the RCs5 group showing the lowest bacterial count (1.46 log CFU/g) and the highest total soluble solid content (10.45%) after 7 days of storage. RCs also reduced weight loss to 34.35% of the control and effectively minimized browning by at least 5.83% compared to the control group. These findings highlight the potential use of RCs to improve the antimicrobial and antioxidant activities of encapsulated phytochemicals, which may have numerous applications in, for example, food preservations.

A bibliometric insight into nanomaterials in vaccine: trends, collaborations, and future avenues.

The emergence of nanotechnology has injected new vigor into vaccine research. Nanovaccine research has witnessed exponential growth in recent years; yet, a comprehensive analysis of related publications has been notably absent. This study utilizes bibliometric methodologies to reveal the evolution of themes and the distribution of nanovaccine research. Using tools such as VOSviewer, CiteSpace, Scimago Graphica, Pajek, R-bibliometrix, and R packages for the bibliometric analysis and visualization of literature retrieved from the Web of Science database. Nanovaccine research commenced in 1981. The publication volume exponentially increased, notably in 2021. Leading contributors include the United States, the Chinese Academy of Sciences, the "Vaccine", and researcher Zhao Kai. Other significant contributors comprise China, the University of California, San Diego, Veronique Preat, the Journal of Controlled Release, and the National Natural Science Foundation of China. The USA functions as a central hub for international cooperation. Financial support plays a pivotal role in driving research advancements. Key themes in highly cited articles include vaccine carrier design, cancer vaccines, nanomaterial properties, and COVID-19 vaccines. Among 7402 keywords, the principal nanocarriers include Chitosan, virus-like particles, gold nanoparticles, PLGA, and lipid nanoparticles. Nanovaccine is primarily intended to address diseases including SARS-CoV-2, cancer, influenza, and HIV. Clustering analysis of co-citation networks identifies 9 primary clusters, vividly illustrating the evolution of research themes over different periods. Co-citation bursts indicate that cancer vaccines, COVID-19 vaccines, and mRNA vaccines are pivotal areas of focus for current and future research in nanovaccines. "candidate vaccines," "protein nanoparticle," "cationic lipids," "ionizable lipids," "machine learning," "long-term storage," "personalized cancer vaccines," "neoantigens," "outer membrane vesicles," "in situ nanovaccine," and "biomimetic nanotechnologies" stand out as research interest. This analysis emphasizes the increasing scholarly interest in nanovaccine research and highlights pivotal recent research themes such as cancer and COVID-19 vaccines, with lipid nanoparticle-mRNA vaccines leading novel research directions.

Self-Reporting Therapeutic Protein Nanoparticles.

We present a modular strategy to synthesize nanoparticle sensors equipped with dithiomaleimide-based, fluorescent molecular reporters capable of discerning minute changes in interparticle chemical environments based on fluorescence lifetime analysis. Three types of nanoparticles were synthesized with the aid of tailor-made molecular reporters, and it was found that protein nanoparticles exhibited greater sensitivity to changes in the core environment than polymer nanogels and block copolymer micelles. Encapsulation of the hydrophobic small-molecule drug paclitaxel (PTX) in self-reporting protein nanoparticles induced characteristic changes in fluorescence lifetime profiles, detected via time-resolved fluorescence spectroscopy. Depending on the mode of drug encapsulation, self-reporting protein nanoparticles revealed pronounced differences in their fluorescence lifetime signatures, which correlated with burst- vs diffusion-controlled release profiles observed in previous reports. Self-reporting nanoparticles, such as the ones developed here, will be critical for unraveling nanoparticle stability and nanoparticle-drug interactions, informing the future development of rationally engineered nanoparticle-based drug carriers.

Proteinaceous Nanoshells with Quasicrystalline Local Order

Among various proteinaceous nanocontainers and nanoparticles, the most promising ones for various applications in nano- and medical science appear to be those whose structures differ fundamentally from icosahedral viral capsids described by the paradigmatic Caspar-Klug model. By analyzing such anomalous assemblies represented in the Protein Data Bank, we identify a series of shells with square-triangular local order and find that most of them originate from short-period approximants of a dodecagonal tiling consisting of square and triangular tiles. Examining the nonequilibrium assembly of such packings, we propose a new method for obtaining periodic square-triangle approximants and then construct the simplest models of tetragonal, octahedral, and icosahedral shells based on cubic and icosahedral nets cut from the approximant structures. Since gluing the nets can change the distances between adjacent vertices of the resulting shell, we introduce an effective energy, the minimization of which equalizes these distances. While the obtained spherical polyhedra reproduce the structures of experimentally observed protein shells and nanoparticles, the principles of protein organization that we lay out, and the ensuing structural models, can help to discover and investigate similar systems in the future. Published by the American Physical Society 2024

The Recent Development of Luteolin-loaded Nanocarrier in Targeting Cancer.

Luteolin (LUT), a naturally occurring flavonoid found in vegetables, fruits, and herbal medicines, has been extensively studied for its pharmacological activities, including anti-proliferative and anticancer effects on various cancer lines. It also exhibits potent antioxidant properties and pro-apoptotic activities against human cancers. However, its therapeutic potential is hindered by its poor solubility in water (5 μg/ml at 45°C) and low bioavailability. This research on the development of luteolin-loaded nanocarrier aims to overcome these limitations, thereby opening up new possibilities in cancer treatment. This paper covers several nanoformulations studied to increase the solubility and bioavailability of LUT. The physicochemical characteristics of the nanoformulation that influence luteolin's solubility and bioavailability have been the subject of more in-depth investigation. Furthermore, it examines how LUT's anti-inflammatory and antioxidant properties aid in lessening the side effects of chemotherapy. Most nanoformulations, including phytosomes, lipid nanoparticles, liposomes, protein nanoparticles, polymer micelles, nanoemulsions, and metal nanoparticles, have shown promising results in improving the solubility and bioavailability of LUT. This is a significant step forward in enhancing the therapeutic potential of LUT in cancer treatment. Furthermore, the study found that LUT's ability to scavenge free radicals can significantly reduce the side effects of cancer treatment, further highlighting its potential to improve patient outcomes. Nanoformulations, because of their unique surface and physiochemical properties, improve the solubility and bioavailability of LUT. However, poor in-vitro and in-vivo correlation and scalability of nanoformulations need to be addressed to achieve good clinical performance of LUT in oncology.

Modulation of rapeseed protein nanoparticles by interaction with chitosan: Enhancing the physical stability, antioxidant activity and β-carotene slow-release properties of Pickering emulsion

This study aimed to investigate the physical stability, oxidation, digestibility and β-carotene sustained-release properties of Pickering emulsions stabilized by rapeseed protein (RP) complexed with chitosan (CS). The results showed that adding CS to RP decreased the RP mean particle size from 3575.0 to 130.9 nm, increased its zeta-potential from 29.6 to 41.4 mV, changed its tertiary structure, and enabled its uniform dispersion. Additionally, CS addition improved RP interfacial properties at air/water and oil/water interfaces, thereby enhancing its emulsification activity and stability. Rheological property analysis exhibited that RP/CS complex had a smaller storage modulus (G′) and loss modulus (G") as well as a higher apparent viscosity. Moreover, the RP/CS complex stabilized emulsions showed a smaller average particle size of 100–1500 nm, a higher zeta-potential of 58–66 mV and better physical stability, thus better tolerating environmental factors such as time, temperature and salt ions. Furthermore, the RP/CS complex stabilized emulsions showed better antioxidant properties with a lower peroxide value (POV) and thiobarbituric acid reactant value (TBARS). Finally, the RP/CS complex emulsions exhibited better antidigestive stability and could slow down the β-carotene release rate. These results provide a theoretical basis for the application of RP/CS complex as a new food emulsifier in the nanocolloid transport system.

Effect of Rice Bran Protein on the Foaming Properties and Foaming Characteristics of Rice Bran Protein-Sodium Caseinate and Rice Bran Protein Nanoparticles-Sodium Caseinate.

Rice bran, a byproduct of rice milling, comprises 12-14% protein. The foaming properties and associated mechanisms of the composite rice bran protein system were not well studied. In this study, a composite protein system composed of rice bran protein (RBP)-sodium caseinate (NaCas) and rice bran protein nanoparticles (RBPNs)-sodium caseinate (NaCas) was investigated. The results showed that the synergistic effect of RBP and NaCas increased the foaming stability of the composite solution up to 83.77 ± 2.75%. Moreover, the foaming capacity and foaming stability of the RBPNs-NaCas composite solution were up to 177.50 ± 3.53% and 80.28 ± 0.39%, respectively. The physicochemical properties results revealed that the particle size volume peaks of RBP-NaCas and RBPNs-NaCas were mainly concentrated at 55.7 nm and 197.1 nm, and RBPNs-NaCas showed a wider single peak particle size distribution. The ζ-potential values of RBP-NaCas and RBPNs-NaCas were changed to -35.5 ± 0.07 mV and -27.2 ± 0.28 mV after complexation. The apparent viscosity and consistency factor of RBP-NaCas decreased by 31.1% compared to RBP, while RBPNs-NaCas displayed similar parameters to the single proteins. The interfacial rheological test showed that RBP and RBPNs can significantly improve the interfacial properties of NaCas by enhancing the interfacial interaction and the interfacial viscoelastic modulus of composite proteins, which is conducive to the stability of the foam system. The outcome of the study provided a theoretical basis for RBP and RBPNs to partially replace NaCas in the processing of foamed food.

Computationally designed mRNA-launched protein nanoparticle vaccines.

Both protein nanoparticle and mRNA vaccines were clinically de-risked during the COVID-19 pandemic1-6. These vaccine modalities have complementary strengths: antigen display on protein nanoparticles can enhance the magnitude, quality, and durability of antibody responses7-10, while mRNA vaccines can be rapidly manufactured11 and elicit antigen-specific CD4 and CD8 T cells12,13. Here we leverage a computationally designed icosahedral protein nanoparticle that was redesigned for optimal secretion from eukaryotic cells14 to develop an mRNA-launched nanoparticle vaccine for SARS-CoV-2. The nanoparticle, which displays 60 copies of a stabilized variant of the Wuhan-Hu-1 Spike receptor binding domain (RBD)15, formed monodisperse, antigenically intact assemblies upon secretion from transfected cells. An mRNA vaccine encoding the secreted RBD nanoparticle elicited 5- to 28-fold higher levels of neutralizing antibodies than an mRNA vaccine encoding membrane-anchored Spike, induced higher levels of CD8 T cells than the same immunogen when delivered as an adjuvanted protein nanoparticle, and protected mice from vaccine-matched and -mismatched SARS-CoV-2 challenge. Our data establish that delivering protein nanoparticle immunogens via mRNA vaccines can combine the benefits of each modality and, more broadly, highlight the utility of computational protein design in genetic immunization strategies.

Effect of Nanoparticle Curvature on Its Interaction with Serum Proteins.

The size or the curvature of nanoparticles (NPs) plays an important role in regulating the composition of the protein corona. However, the molecular mechanisms of how curvature affects the interaction of NPs with serum proteins still remain elusive. In this study, we employ all-atom molecular dynamics simulations to investigate the interactions between two typical serum proteins and PEGylated Au NPs with three different surface curvatures (0, 0.1, and 0.5 nm-1, respectively). The results show that for proteins with a regular shape, the binding strength between the serum protein and Au NPs decreases with increasing curvature. For irregularly shaped proteins with noticeable grooves, the binding strength between the protein and Au NPs does not change obviously with increasing curvature in the cases of smaller curvature. However, as the curvature continues to increase, Au NPs may act as ligands firmly adsorbed in the protein grooves, significantly enhancing the binding strength. Overall, our findings suggest that the impact of NP curvature on protein adsorption may be nonmonotonic, which may provide useful guidelines for better design of functionalized NPs in biomedical applications.

Heterologous Prime-Boost with Immunologically Orthogonal Protein Nanoparticles for Peptide Immunofocusing.

Protein nanoparticles are effective platforms for antigen presentation and targeting effector immune cells in vaccine development. Encapsulins are a class of protein-based microbial nanocompartments that self-assemble into icosahedral structures with external diameters ranging from 24 to 42 nm. Encapsulins from Myxococcus xanthus were designed to package bacterial RNA when produced in E. coli and were shown to have immunogenic and self-adjuvanting properties enhanced by this RNA. We genetically incorporated a 20-mer peptide derived from a mutant strain of the SARS-CoV-2 receptor binding domain (RBD) into the encapsulin protomeric coat protein for presentation on the exterior surface of the particle, inducing the formation of several nonicosahedral structures that were characterized by cryogenic electron microscopy. This immunogen elicited conformationally relevant humoral responses to the SARS-CoV-2 RBD. Immunological recognition was enhanced when the same peptide was presented in a heterologous prime/boost vaccination strategy using the engineered encapsulin and a previously reported variant of the PP7 virus-like particle, leading to the development of a selective antibody response against a SARS-CoV-2 RBD point mutant. While generating epitope-focused antibody responses is an interplay between inherent vaccine properties and B/T cells, here we demonstrate the use of orthogonal nanoparticles to fine-tune the control of epitope focusing.

In Situ Measurement of Nanoparticle-Blood Protein Adsorption and Its Heterogeneity with Single-Nanoparticle Resolution via Dual Fluorescence Quantification.

The formation of a protein corona gives nanomedicines a distinct biological identity, profoundly influencing their fate in the body. Nonspecific nanoparticle-protein interactions are typically highly heterogeneous, which can lead to unique biological behaviors and in vivo fates for individual nanoparticles that remain underexplored. To address this, we have established an in situ approach that allows quantitative examination of nanoparticle-protein adsorption at the individual nanoparticle level. This method integrates dual fluorescence quantification techniques, wherein the nanoparticles are first individually analyzed via nanoflow cytometry to detect fluorescent signals from adsorbed proteins. The obtained fluorescence intensity is then translated into protein quantities through calibration with microplate reader quantification. Consequently, this approach enables analysis of interparticle heterogeneity of nano-protein interactions, as well as in situ monitoring of protein adsorption kinetics and nanoparticle aggregation status in blood serum, preconditioning for a comprehensive understanding of nano-bio interactions, and predicting in vivo fate of nanomedicines.

The formation of protein coronas and its effect on the quercetin-edible dock protein nanoparticles

Nanoparticles enter into the digestive tract will quickly adsorb the protein-based biomacromolecules and form protein coronas, which can affect the release characteristic of the encapsulated bioactive compounds. In this study, edible dock protein (EDP) nanoparticles were chosen as the model nanoparticles, and quercetin (Que) was chosen as the representative bioactive compound, the regulatory mechanism of protein coronas on the release characteristic of Que within the EDP nanoparticles was investigated. Results showed that the digestive enzymes can adsorb on the surface of the Que-EDP nanoparticles and form protein coronas during the simulated digestion. Herein, hydrogen bonding, van der Waals force, and hydrophobic interactions were the main adsorption forces between digestive enzymes and nanoparticles. Moreover, the protein coronas formed by α-amylase, pepsin, trypsin, and amylopsin increased the zeta potential of the Que-EDP nanoparticles from −17.9 mV to −11.2 mV, +3.8 mV to +7.0 mV, −21.9 mV to −19.2 mV, and −21.9 mV to −12.1 mV, respectively. The particle size, polydispersity index, and protein conformation of the nanoparticles were also remarkably changed after the formation of protein coronas. Besides, the activity of α-amylase and amylopsin on the surface of the Que-EDP nanoparticles was enhanced by 1.4 and 1.6 times. The simulated digestion experiments indicated that α-amylase and amylopsin coronas slowed the release of Que, while pepsin and trypsin coronas accelerated the release of Que. This study will provide a new idea and theoretical basis for the rational design of nanoparticle carriers.

Rapid Plasma Proteome Profiling via Nanoparticle Protein Corona and Direct Infusion Mass Spectrometry.

Noninvasive detection of protein biomarkers in plasma is crucial for clinical purposes. Liquid chromatography-mass spectrometry (LC-MS) is the gold standard technique for plasma proteome analysis, but despite recent advances, it remains limited by throughput, cost, and coverage. Here, we introduce a new hybrid method that integrates direct infusion shotgun proteome analysis (DISPA) with nanoparticle (NP) protein corona enrichment for high-throughput and efficient plasma proteomic profiling. We realized over 280 protein identifications in 1.4 min collection time, which enables a potential throughput of approximately 1000 samples daily. The identified proteins are involved in valuable pathways, and 44 of the proteins are FDA-approved biomarkers. The robustness and quantitative accuracy of this method were evaluated across multiple NPs and concentrations with a mean coefficient of variation of 17%. Moreover, different protein corona profiles were observed among various NPs based on their distinct surface modifications, and all NP protein profiles exhibited deeper coverage and better quantification than neat plasma. Our streamlined workflow merges coverage and throughput with precise quantification, leveraging both DISPA and NP protein corona enrichment. This underscores the significant potential of DISPA when paired with NP sample preparation techniques for plasma proteome studies.

Enhancing cross-protection against influenza by heterologous sequential immunization with mRNA LNP and protein nanoparticle vaccines

Enhancing influenza vaccine cross-protection is imperative to alleviate the significant public health burden of influenza. Heterologous sequential immunization may synergize diverse vaccine formulations and routes to improve vaccine potency and breadth. Here we investigate the effects of immunization strategies on the generation of cross-protective immune responses in female Balb/c mice, utilizing mRNA lipid nanoparticle (LNP) and protein-based PHC nanoparticle vaccines targeting influenza hemagglutinin. Our findings emphasize the crucial role of priming vaccination in shaping Th bias and immunodominance hierarchies. mRNA LNP prime favors Th1-leaning responses, while PHC prime elicits Th2-skewing responses. We demonstrate that cellular and mucosal immune responses are pivotal correlates of cross-protection against influenza. Notably, intranasal PHC immunization outperforms its intramuscular counterpart in inducing mucosal immunity and conferring cross-protection. Sequential mRNA LNP prime and intranasal PHC boost demonstrate optimal cross-protection against antigenically drifted and shifted influenza strains. Our study offers valuable insights into tailoring immunization strategies to optimize influenza vaccine effectiveness.