Human ferritin is regarded as an attractive and promising vaccine platform because of its uniform structure, good plasticity, and desirable thermal and chemical stabilities. Besides, it is biocompatible and presumed safe when used as a vaccine carrier. However, there is a lack of knowledge of how different antigen insertion sites on the ferritin nanocage impact the resulting protein stability and performance. To address this question, we selected Epstein–Barr nuclear antigen 1 as a model epitope and fused it at the DNA level with different insertion sites, namely, the N- and C-termini of ferritin, to engineer proteins E1F1 and F1E1, respectively. Protein properties including hydrophobicity and thermal, pH, and chemical stability were investigated both by molecular dynamics (MD) simulation and by experiments. Both methods demonstrate that the insertion site plays an important role in protein properties. The C-terminus insertion (F1E1) leads to a less hydrophobic surface and more tolerance to the external influence of high temperature, pH, and high concentration of chemical denaturants compared to N-terminus insertion (E1F1). Simulated protein hydrophobicity and thermal stability by MD were in high accordance with experimental results. Thus, MD simulation can be used as a valuable tool to engineer nanovaccine candidates, cutting down costs by reducing the experimental effort and accelerating vaccine design.
Ferritin, one of the most investigated protein nanocages, is considered as a promising drug carrier because of its advantageous stability and safety. However, its short half-life and undesirable tumor targeting ability has limited its usage in tumor treatment. In this work, two types of functional peptides, half-life extension peptide PAS, and tumor targeting peptide RGDK (Arg-Gly-Asp-Lys), are inserted to human heavy-chain ferritin (HFn) at C-terminal through flexible linkers with two distinct enzyme cleavable sites. Structural characterizations show both HFn and engineered HFns can assemble into nanoparticles but with different apparent hydrodynamic volumes and molecular weights. RGDK peptide enhanced the internalization efficiency of HFn and showed a significant increase of growth inhibition against 4T1 cell line in vitro. Pharmacokinetic study in vivo demonstrates PAS peptides extended ferritin half-life about 4.9 times in Sprague Dawley rats. RGDK peptides greatly enhanced drug accumulation in the tumor site rather than in other organs in biodistribution analysis. Drug loaded PAS-RGDK functionalized HFns curbed tumor growth with significantly greater efficacies in comparison with drug loaded HFn.
Whey is a by-product of cheese, casein, and yogurt manufacture. It contains a mixture of proteins that need to be isolated and purified to fully exploit their nutritional and functional characteristics. Protein-enriched fractions and highly purified proteins derived from whey have led to the production of valuable ingredients for many important food and pharmaceutical applications. This article provides a review on the separation principles behind both the commercial and emerging techniques used for whey protein fractionation, as well as the efficacy and limitations of these techniques in isolating and purifying individual whey proteins. The fractionation of whey proteins has mainly been achieved at commercial scale using membrane filtration, resin-based chromatography, and the integration of multiple technologies (e.g., precipitation, membrane filtration, and chromatography). Electromembrane separation and membrane chromatography are two main emerging techniques that have been developed substantially in recent years. Other new techniques such as aqueous two-phase separation and magnetic fishing are also discussed, but only a limited number of studies have reported their application in whey protein fractionation. This review offers useful insights into research directions and technology screening for academic researchers and dairy processors for the production of whey protein fractions with desired nutritional and functional properties.
Human heavy-chain ferritin (HFn) and hepatitis B virus core (HBc) are both nanoparticle proteins presenting a well-oriented architecture with constant size and shape, which can be engineered to carry epitopes on the surface of the nanoparticle protein cage, enabling vaccine design. This study aims to investigate the immunogenicity differences between engineered HFn and chimeric HBc bearing the same epitope. As a proof of concept, the model epitope Epstein−Barr nuclear antigen 1 (EBNA1) is inserted at the N-terminus of the HFn and HBc subunit to produce two vaccine candidates named EBNA1−HFn (E1F1) and EBNA1−HBc (E1H1), respectively. From in vivo immunogenicity studies, E1H1 demonstrates the capability to prompt significant humoral and cell-mediated immune responses in adjuvant-free formulation. When formulated with the aluminum hydroxide adjuvant, E1H1 produces approximately 5× higher titer and 2× stronger proliferation index (PI) than E1F1. These results confirm that the HBc carrier induces a stronger humoral immune response than HFn. On the other hand, from lymphocyte activation experiments, E1F1 induces a stronger cell-mediated immune response indicated by 5× more CD8 + T cells and 2× more effector memory T cells in the E1F1 group versus the E1H1 group. Through this study, HFn and HBc are shown to be potentially effective vaccine carrier nanoparticles having subtly different immunological responses. KEYWORDS: protein-based vaccine molecule design, hepatitis B virus core, virus like particles, human ferritin heavy chain, Epstein−Barr nuclear antigen 1
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.