Development of a microchip capillary electrophoresis method for determination of the purity and integrity of mRNA in lipid nanoparticle vaccines

Messenger RNA (mRNA)-based vaccines are advantageous because they can be relatively quicker and more cost efficient to manufacture compared to other traditional vaccine products. Lipid nanoparticles have three common purposes: delivery, self-adjuvanting properties, and mRNA protection. Faster vaccine development requires an efficient and fast assay to monitor mRNA purity and integrity. Microchip CE is known to be a robust technology that is capable of rapid separation. Here, we describe the development and optimization of a purity and integrity assay for mRNA-based vaccines encapsulated in lipid nanoparticles using commercial microchip-based separation. The analytical parameters of the optimized assay were assessed and the method is a stability indicating assay.     

Maltodextrin as stabilizer for emulsion polymerization: Adsorption and grafting behavior

Emulsion polymerization of the three-monomer system butyl acrylate–styrene–methacrylic acid was performed in batch using a commercial maltodextrin derived from starch degradation as stabilizer. Stable latexes with narrow particle size distributions were obtained in all examined cases. A method was developed to analyze and quantify the partitioning of the maltodextrin between the continuous phase (supernatant) and the particle phase. Significant differences between the polysaccharides adsorbed onto particles with or without emulsion polymerization reaction were observed. The possible reactions of maltodextrin in presence of a radical initiator were studied in aqueous phase, thus confirming maltodextrin degradation. The formation of copolymers involving the original monomers and the stabilizer according to two different reactive pathways was also confirmed. In terms of adsorbed maltodextrin, two different contributions were observed: maltodextrin physically adsorbed and maltodextrin chemically grafted and/or physically incorporated into the polymer.     

Blue-LED Synthesis of Pummerer's Ketones by Peroxidase-Lignin-Catalyzed Oxidative Coupling of Substituted Phenols

Pummerer's ketones resembling the tricyclic scaffold of bioactive natural substances were synthesized by blue-LED driven Horseradish Peroxidase oxidative coupling of substituted phenols in 2-methyltetrahydrofuran by using meso-tetraphenylporphyrin as photosensitizer and dioxygen as primary oxidant. The application of functionalized lignin nanoparticles as a renewable and efficient platform for the immobilization of the enzyme extended the effectiveness of the overall process to heterogeneous catalysis under buffer limiting conditions.     

Phase Behavior of a Designed Cyclopropyl Analogue of Monoolein: Implications for Low‐Temperature Membrane Protein Crystallization

Lipidic cubic phases (LCPs) are used in areas ranging from membrane biology to biodevices. Because some membrane proteins are notoriously unstable at room temperature, and available LCPs undergo transformation to lamellar phases at low temperatures, development of stable low‐temperature LCPs for biophysical studies of membrane proteins is called for. Monodihydrosterculin (MDS) is a designer lipid based on monoolein (MO) with a configurationally restricted cyclopropyl ring replacing the olefin. Small‐angle X‐ray scattering (SAXS) analyses revealed a phase diagram for MDS lacking the high‐temperature, highly curved reverse hexagonal phase typical for MO, and extending the cubic phase boundary to lower temperature, thereby establishing the relationship between lipid molecular structure and mesophase behavior. The use of MDS as a new material for LCP‐based membrane protein crystallization at low temperature was demonstrated by crystallizing bacteriorhodopsin at 20 °C as well as 4 °C.     

Fibrillar Networks of Glycyrrhizic Acid for Hybrid Nanomaterials with Catalytic Features

Self‐assembly of the naturally occurring sweetening agent, glycyrrhizic acid (GA) in water is studied by small‐angle X‐ray scattering and microscopic techniques. Statistical analysis on atomic force microscopy images reveals the formation of ultralong GA fibrils with uniform thickness of 2.5 nm and right‐handed twist with a pitch of 9 nm, independently of GA concentration. Transparent nematic GA hydrogels are exploited to create functional hybrid materials. Two‐fold and three‐fold hybrids are developed by introducing graphene oxide (GO) and in situ‐synthesized gold nanoparticles (Au NPs) in the hydrogel matrix for catalysis applications. In the presence of GO, the catalytic efficiency of Au NPs in the reduction of p‐nitrophenol to p‐aminophenol is enhanced by 2.5 times. Gold microplate single crystals are further synthesized in the GA hydrogel, expanding the scope of these hybrids and demonstrating their versatility in materials design.     

An exact algorithm to extend lifetime through roles allocation in sensor networks with connectivity constraints

We face the problem of scheduling optimally the activities in a wireless sensor network in order to ensure that, in each instant of time, the activated sensors can monitor all points of interest (targets) and route the collected information to a processing facility. Each sensor is allocated to a role, depending on whether it is actually used to monitor the targets, to forward information or kept idle, leading to different battery consumption ratios. We propose a column generation algorithm that embeds a highly efficient genetic metaheuristic for the subproblem. Moreover, to optimally solve the subproblem, we introduce a new formulation with fewer integer variables than a previous one proposed in the literature. Finally, we propose a stopping criterion to interrupt the optimal resolution of the subproblem as soon as a favorable solution is found. The results of our computational tests show that our algorithm consistently outperforms previous approaches in the literature, and also improves the best results known to date on some benchmark instances.     

Conjugate Direction Methods and Polarity for Quadratic Hypersurfaces

We use some results from polarity theory to recast several geometric properties of Conjugate Gradient-based methods, for the solution of nonsingular symmetric linear systems. This approach allows us to pursue three main theoretical objectives. First, we can provide a novel geometric perspective on the generation of conjugate directions, in the context of positive definite systems. Second, we can extend the above geometric perspective to treat the generation of conjugate directions for handling indefinite linear systems. Third, by exploiting the geometric insight suggested by polarity theory, we can easily study the possible degeneracy (pivot breakdown) of Conjugate Gradient-based methods on indefinite linear systems. In particular, we prove that the degeneracy of the standard Conjugate Gradient on nonsingular indefinite linear systems can occur only once in the execution of the Conjugate Gradient.