ace2  (BPS Bioscience)

Journal: Journal of Virology

Article Title: Targeting envelope lipids with 2,6‑di‑ O ‑methyl‑3‑acetyl‑β‑cyclodextrin impairs infectivity of SARS‑CoV‑2 and Japanese encephalitis virus

doi: 10.1128/jvi.01357-25

Figure Lengend Snippet: DMA-β-CyD acts directly on SARS-CoV-2 and reduces its infectivity titer. DMA-β-CyD was directly exposed to different SARS-CoV-2 variants for ( A ) 10 min and ( B ) 180 min. Viral infectivity titers were expressed in TCID 50 /mL, and significant differences (Student’s t -test) are indicated in the figures. Mean values from at least three independent experiments are shown, and error bars indicate SD. ( C ) Competitive inhibition experiment with DMA-β-CyD. Recombinant ACE2 was used, and DMA-β-CyD was pre-incubated before the loading of the recombinant spike protein. Statistical significance was calculated using one-way analysis of variance (ANOVA) followed by Dunnett’s test. n.s. indicates not significant. The mean values of at least three independent experiments are shown. The error bars denote SD.

Article Snippet: To determine whether DMA-β-CyD inhibits the binding of the SARS-CoV-2 spike protein to ACE2, an ACE2:SARS-CoV-2 Spike S1 Inhibitor Screening Assay Kit (BPS Bioscience, Inc.) was used.

Techniques: Infection, Inhibition, Recombinant, Incubation

Journal: Journal of Virology

Article Title: Targeting envelope lipids with 2,6‑di‑ O ‑methyl‑3‑acetyl‑β‑cyclodextrin impairs infectivity of SARS‑CoV‑2 and Japanese encephalitis virus

doi: 10.1128/jvi.01357-25

Figure Lengend Snippet: DMA-β-CyD prolongs the virus-induced reduction in clotting time in human serum by masking aminophospholipids in the viral lipid envelope. ( A ) The Wuhan, Beta, and Delta variants prepared in Vero E6/TMPRSS2 cells were treated with DMA-β-CyD for the indicated times, followed by a clotting assay. ( B ) The Wuhan variant, prepared in A549 cells overexpressing ACE2, was treated with DMA-β-CyD for the indicated times, followed by a clotting assay. ( C ) To examine the significance of the molecular encapsulation activity of DMA-β-CyD, pretreatment with 10 mM DMAβCyD with equimolar (10 mM) 1adamantanecarboxylic acid was performed to assess its effect on the virus-induced reduction in clotting time. ( D ) The Wuhan variant, prepared in Vero E6/TMPRSS2 cells, was treated with DM-β-CyD for the indicated times, followed by a clotting assay. P values were determined using one-way ANOVA (Dunnett’s test). * indicates a statistically significant difference in clotting time between groups untreated and treated with DMA-β-CyD or DM-β-CyD ( P < 0.05). Data represent the means of at least three independent experiments, and error bars indicate SD.

Article Snippet: To determine whether DMA-β-CyD inhibits the binding of the SARS-CoV-2 spike protein to ACE2, an ACE2:SARS-CoV-2 Spike S1 Inhibitor Screening Assay Kit (BPS Bioscience, Inc.) was used.

Techniques: Virus, Coagulation, Variant Assay, Encapsulation, Activity Assay

Journal: Journal of Virology

Article Title: Targeting envelope lipids with 2,6‑di‑ O ‑methyl‑3‑acetyl‑β‑cyclodextrin impairs infectivity of SARS‑CoV‑2 and Japanese encephalitis virus

doi: 10.1128/jvi.01357-25

Figure Lengend Snippet: Inhibition of viral entry by DMA-β-CyD and evaluation of viral particle stability. ( A ) DMA-β-CyD was preincubated with the SARS-CoV-2 Wuhan variant, which was prepared in Vero E6/TMPRSS2 cells. Free DMA-β-CyD was removed by gel filtration, and the resulting viral particles were then used to infect cells. Four hours after infection, total RNA was extracted from Vero E6/TMPRSS2 cells, and gRNA was detected by RT-PCR. P values were determined by one-way ANOVA using Dunnett’s test (** P < 0.01). N.D. indicates that the SARS-CoV-2 phenotypic RNA (gRNA) was not detected. Data represent the means of at least three independent experiments, and error bars indicate SD. ( B ) DMA-β-CyD was preincubated with the SARS-CoV-2 Wuhan variant, which was prepared in A549 cells overexpressing ACE2. The efficiency of viral entry was similarly examined. Western blot analysis results, shown in the bar graph inset, confirm ACE2 expression in A549 cells. ( C ) The SARS-CoV-2 Wuhan variant, prepared in Vero E6/TMPRSS2 cells, was treated with anti-SARS-CoV-2 patient serum or DMA-β-CyD for 180 min, followed by virus purification by ultracentrifugation and western blot analysis using patient antiserum. ( D ) Evaluation of virus particle stability. (Left panel) The SARS-CoV-2 Wuhan variant, prepared in Vero E6/TMPRSS2 cells, was treated with an anti-S2 antibody and then subjected to gel filtration. Gel filtration was also performed using mouse IgG and IgM as molecular weight markers. The resulting flow-through fraction was analyzed by western blot analysis. (Right panel) The SARS-CoV-2 Wuhan variant, prepared in Vero E6/TMPRSS2 cells, was treated with DMA-β-CyD or anti-SARS-CoV-2 serum. The virus was purified by gel filtration, captured on a plate coated with anti-RBD antibodies, and the number of plate-bound viral particles was quantified by RT-qPCR. P values were determined by one-way ANOVA using Dunnett’s test (** P < 0.01). N.D. indicates that no detectable SARS-CoV-2 gRNA was found. Data represent the means of at least three independent experiments, and error bars indicate SD.

Article Snippet: To determine whether DMA-β-CyD inhibits the binding of the SARS-CoV-2 spike protein to ACE2, an ACE2:SARS-CoV-2 Spike S1 Inhibitor Screening Assay Kit (BPS Bioscience, Inc.) was used.

Techniques: Inhibition, Variant Assay, Filtration, Infection, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Virus, Purification, Molecular Weight, Quantitative RT-PCR