RESUMEN
Neutralizing antibodies (NAbs) hold great promise for clinical interventions against SARS-CoV- 2 variants of concern (VOCs). Understanding NAb epitope-dependent antiviral mechanisms is crucial for developing vaccines and therapeutics against VOCs. Here we characterized two potent NAbs, EH3 and EH8, isolated from an unvaccinated pediatric patient with exceptional plasma neutralization activity. EH3 and EH8 cross-neutralize the early VOCs and mediate strong Fc-dependent effector activity in vitro. Structural analyses of EH3 and EH8 in complex with the receptor-binding domain (RBD) revealed the molecular determinants of the epitope-driven protection and VOC-evasion. While EH3 represents the prevalent IGHV3-53 NAb whose epitope substantially overlaps with the ACE2 binding site, EH8 recognizes a narrow epitope exposed in both RBD-up and RBD-down conformations. When tested in vivo, a single-dose prophylactic administration of EH3 fully protected stringent K18-hACE2 mice from lethal challenge with Delta VOC. Our study demonstrates that protective NAbs responses converge in pediatric and adult SARS-CoV-2 patients.
RESUMEN
For the first time, a magnetic solid-phase extraction with high-performance liquid chromatography detection method using Zr functionalized Fe3 O4 magnetic material to enrich ribavirin was successfully established. Zr components that modified in Fe3 O4 nanoparticles via a simple one-step hydrothermal method was selected in this work to specifically capture ribavirin by the strong chemical bonding between Zr components of Zr functionalized Fe3 O4 magnetic material and cis-hydroxyl of ribavirin, which was confirmed by pseudo-second-order kinetic model. And Fe3 O4 components were selected in this work to achieve simple operation. Under the optimal experimental conditions, proposed magnetic solid-phase extraction with high-performance liquid chromatography detection method along with Zr functionalized Fe3 O4 magnetic material offered a wide range linearity at 10-200 µg/L with correlation coefficient of 0.9978 with low detection limit of 2.68 µg/L for ribavirin. The relative standard deviations obtained from nine parallel extractions of 100 µg/L ribavirin were 4.41% and revealed good repeatability. This established method was successfully applied to detect real samples including chicken liver, egg, and shrimp with satisfactory recoveries of 74.13-92.9%.
Asunto(s)
Alimentación Animal/análisis , Óxido Ferrosoférrico/química , Contaminación de Alimentos/análisis , Ribavirina/análisis , Extracción en Fase Sólida , Circonio/química , Cromatografía Líquida de Alta Presión , Fenómenos Magnéticos , Estructura Molecular , Tamaño de la PartículaRESUMEN
The F-BAR domain containing proteins PACSINs are cytoplasmic phosphoproteins involved in various membrane deformations, such as actin reorganization, vesicle transport and microtubule movement. Our previous study shows that all PACSINs are composed of crescent shaped dimers with two wedge loops, and the wedge loop-mediated lateral interaction between neighboring dimers is important for protein packing and tubulation activity. Here, from the crystal packing of PACSIN 2, we observed a tight tip-to-tip interaction, in addition to the wedge loop-mediated lateral interaction. With this tip-to-tip interaction, the whole packing of PACSIN 2 shows a spiral-like assembly with a central hole from the top view. Elimination of this tip-to-tip connection inhibited the tubulation function of PACSIN 2, indicating that tip-to-tip interaction plays an important role in membrane deformation activity. Together with our previous study, we proposed a packing model for the assembly of PACSIN 2 on membrane, where the proteins are connected by tip-to-tip and wedge loop-mediated lateral interactions on the surface of membrane to generate various diameter tubules.