Capture and neutralization of SARS-CoV-2 and influenza virus by algae-derived lectins with high-mannose and core fucose specificities.

We first investigated the interactions between several algae-derived lectins and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). We created lectin columns using high-mannose (HM)-type glycan-specific lectins OAA and KAA-1 or core fucose-specific lectin hypninA-2 and conducted binding experiments with SARS-CoV-2. The results showed that these lectins were capable of binding to the virus. Furthermore, when examining the neutralization ability of nine different lectins, it was found that KAA-1, ESA-2, and hypninA-2 were effective in neutralizing SARS-CoV-2. In competitive inhibition experiments with glycoproteins, neutralization was confirmed to occur through HM-type or core fucose-type glycans. However, neutralization was not observed with other lectins, such as OAA. This trend of KAA-1 and ESA-2 having the neutralizing ability and OAA not having it was also similar to influenza viruses. Electron microscopy observations revealed that KAA-1 and hypninA-2 strongly aggregated SARS-CoV-2 particles, while OAA showed a low degree of aggregation. It is believed that the neutralization of SARS-CoV-2 involves multiple factors, such as glycan attachment sites on the S protein, the size of lectins, and their propensity to aggregate, which cause inhibition of receptor binding or aggregation of virus particles. This study demonstrated that several algae-derived lectins could neutralize SARS-CoV-2 and that lectin columns can effectively recover and concentrate the virus.

Structural basis of spike RBM-specific human antibodies counteracting broad SARS-CoV-2 variants.

The decrease of antibody efficacy to mutated SARS-CoV-2 spike RBD explains the breakthrough infections and reinfections by Omicron variants. Here, we analyzed broadly neutralizing antibodies isolated from long-term hospitalized convalescent patients of early SARS-CoV-2 strains. One of the antibodies named NCV2SG48 is highly potent to broad SARS-CoV-2 variants including Omicron BA.1, BA.2, and BA.4/5. To reveal the mode of action, we determined the sequence and crystal structure of the Fab fragment of NCV2SG48 in a complex with spike RBD from the original, Delta, and Omicron BA.1. NCV2SG48 is from a minor VH but the multiple somatic hypermutations contribute to a markedly extended binding interface and hydrogen bonds to interact with conserved residues at the core receptor-binding motif of RBD, which efficiently neutralizes a broad spectrum of variants. Thus, eliciting the RBD-specific B cells to the longitudinal germinal center reaction confers potent immunity to broad SARS-CoV-2 variants emerging one after another.

Thermal Structure Transformation of Methylammonium Vanadate and it's Application as a Negative Staining Reagent for Observing SARS‐CoV‐2

The solid‐state thermal structure transformation of methylammonium vanadate, (CH3NH3)VO3, from −150 °C to 350 °C is reported. Variable‐temperature X‐ray single‐crystal structure analysis at 23, 0, −50, −100, and −150 °C reveal (CH3NH3)VO3 comprises of methylammonium cations and “snake‐like” ([VO3]−)n anion chains propagating along the c‐direction in the Pna21 space group. In between −150 and −100 °C, we observe a reversible structural transformation due to the re‐orientation of the methylammonium cations in the crystal packing, which is also confirmed by the reversible profiles observed in differential scanning calorimetry. The methylammonium vanadate is stable until at ca. 100 °C and further heating releases methylamine and water and V2O5 is formed at ca. 275 °C . Furthermore, we show that the methylammonium vanadate can be used as a negative staining reagent for visualizing SARS‐CoV‐2, allowing us to discern the spike proteins from the body of the virus using transmission electron microscopy. Methylammonium vanadate, [(CH3NH3)VO3]n, shows a reversible structure transformation due to change of methylammonium cation orientation between −100 and −150 °C. In contrast, heating causes decomposition to form V2O5. The methylammonium vanadate can be used as a negative staining reagent for visualizing SARS‐CoV‐2 using transmission electron microscopy.

Thermal Structure Transformation of Methylammonium Vanadate and it's Application as a Negative Staining Reagent for Observing SARS-CoV-2.

The solid-state thermal structure transformation of methylammonium vanadate, (CH3NH3)VO3, from -150 °C to 350 °C is reported. Variable-temperature X-ray single-crystal structure analysis at 23, 0, -50, -100, and -150 °C reveal (CH3NH3)VO3 comprises of methylammonium cations and "snake-like" ([VO3]-)n anion chains propagating along the c-direction in the Pna21 space group. In between -150 and -100 °C, we observe a reversible structural transformation due to the re-orientation of the methylammonium cations in the crystal packing, which is also confirmed by the reversible profiles observed in differential scanning calorimetry. The methylammonium vanadate is stable until at ca. 100 °C and further heating releases methylamine and water and V2O5 is formed at ca. 275 °C . Furthermore, we show that the methylammonium vanadate can be used as a negative staining reagent for visualizing SARS-CoV-2, allowing us to discern the spike proteins from the body of the virus using transmission electron microscopy.