Journal: iScience

Article Title: Soluble ACE2 correlates with severe COVID-19 and can impair antibody responses

doi: 10.1016/j.isci.2024.109330

Figure Lengend Snippet:

Article Snippet: Anti-human ACE2 antibody F49433 , NSJ Bioreagents , #F49433.

Techniques: Recombinant, Magnetic Beads, Software, In Silico

Journal: Pharmaceuticals

Article Title: In Vitro Effect of Taraxacum officinale Leaf Aqueous Extract on the Interaction between ACE2 Cell Surface Receptor and SARS-CoV-2 Spike Protein D614 and Four Mutants

doi: 10.3390/ph14101055

Figure Lengend Snippet: Binding inhibition of S1 spike protein to human HEK293-hACE2 cells by extract pre-incubation. Cells were pre-incubated for the indicated times with 10 mg/mL T. officinale (TO), its HMW fraction, equal to 10 mg/mL extract (HMW), and 10 mg/mL C. intybus (CI) or solvent control (a.d.) and subsequently treated with His-tagged S1 spike protein for 1 h without a washing step in between at 4 °C. Binding inhibition was assessed using flow cytometry. N = 3, bars are means + SD. Upper left: cytogram of gated HEK-hACE2 cells. Middle: overlay of representative fluorescence intensity histograms for ACE2 surface expression. Upper right: overlay of representative fluorescence intensity histograms for spike-binding inhibition by the extracts or a.d.; positive control: 20 µg/mL soluble hACE2. Cells were stained with anti-His-tag Alexa Fluor 647 conjugated monoclonal antibody; ** p < 0.01. Significance of difference was calculated relative to the solvent control by one-way ANOVA.

Article Snippet: Anti-hACE2 antibody (Biomol, NSJ-F49433) was used as reference for transduction inhibition assay.

Techniques: Binding Assay, Inhibition, Incubation, Flow Cytometry, Fluorescence, Expressing, Positive Control, Staining

Journal: Pharmaceuticals

Article Title: In Vitro Effect of Taraxacum officinale Leaf Aqueous Extract on the Interaction between ACE2 Cell Surface Receptor and SARS-CoV-2 Spike Protein D614 and Four Mutants

doi: 10.3390/ph14101055

Figure Lengend Snippet: Binding inhibition of spike D614, and its mutants D614G, N501Y or mix (N501Y, K417N and E484K) to human HEK293-hACE2 and A549-hACE2-TMPRSS2 cells by extract pre- or post-incubation. Overlay of fluorescence intensity histogram for ( A ) unstained HEK cells, staining control (anti-His-tag Alexa Fluor 647), and cells incubated with His-tag-labelled spike D614, D614G or N501Y for 1 h at 4 °C. ( B , C ) cells pre-incubated with solvent control (a.d.), 10 mg/mL T. officinale (TO) or 10 mg/mL C. intybus (CI) for 30–60 s, and then treated with His-tag-labelled S1 spike D614, D614G or N501Y protein for 1 h without a washing step in between at 4 °C. ( D – G ) Effect of extract incubation on HEK or A549 cells either before or after incubation with His-tag-labelled spike D614, D614G, N501Y or mix (N501Y, K417N and E484K) protein at 37 °C. ( H ) Plant extracts were incubated in saliva from four human donors for 0.5 h at 37 °C. Afterwards, cells were pre-treated with 5 mg/mL extracts for 60 s at 37 °C before incubation with His-tag-labelled spike D614 protein for 0.5 h at 37 °C. Spike-binding inhibition to human cells was assessed using flow cytometric analysis of cells stained with anti-His-tag Alexa Fluor 647 conjugated monoclonal antibody. Bars are means +SD; * p < 0.05, ** p < 0.01. Significance of difference was calculated relative to the respective solvent control by one-way ANOVA.

Article Snippet: Anti-hACE2 antibody (Biomol, NSJ-F49433) was used as reference for transduction inhibition assay.

Techniques: Binding Assay, Inhibition, Incubation, Fluorescence, Staining

Journal: Pharmaceuticals

Article Title: In Vitro Effect of Taraxacum officinale Leaf Aqueous Extract on the Interaction between ACE2 Cell Surface Receptor and SARS-CoV-2 Spike Protein D614 and Four Mutants

doi: 10.3390/ph14101055

Figure Lengend Snippet: Effect of T. officinale extract on ACE2 enzyme activity and protein expression. ( A ) Viability of A549-hACE2-TMPRSS2 cells was determined using trypan blue cell staining after 84 h exposure to the extract. ( B ) Cells were incubated with TO extract or 500 ng/mL S1 protein and analyzed for enzyme activity using a fluorescence kit. ( C , D ) Cells were exposed for 6 h or 24 h to extract without (white bars) or with (black bars) 500 ng/mL S1 protein and analyzed for ACE2 protein expression using a human ACE2 ELISA kit; a.d.: solvent control. Bars are means + SD, N ≥ 3 independent experiments; * p < 0.05, ** p < 0.01. Significance of difference was calculated relative to the respective control by one-way ANOVA.

Article Snippet: Anti-hACE2 antibody (Biomol, NSJ-F49433) was used as reference for transduction inhibition assay.

Techniques: Activity Assay, Expressing, Staining, Incubation, Fluorescence, Enzyme-linked Immunosorbent Assay

Journal: Pharmaceuticals

Article Title: In Vitro Effect of Taraxacum officinale Leaf Aqueous Extract on the Interaction between ACE2 Cell Surface Receptor and SARS-CoV-2 Spike Protein D614 and Four Mutants

doi: 10.3390/ph14101055

Figure Lengend Snippet: Viral transduction inhibition of A549-hACE2-TMPRSS2 cells by T. officinale extract. ( A ) Cells were pre-treated with T. officinale (TO) or HMW extract for 0.5 h before infection with 7500 TU/mL SARS-CoV-2 spike D614 or Delta (B.1.617.2) variant for 24 h; ( B , C ) Cells were transduced with 7500 TU/mL SARS-CoV-2 for (B) 3 h before addition of TO for another 21 h or (C) 24 h. After transduction, the medium was exchanged with fresh medium containing TO or HMW extract at the indicated concentrations and post-incubated for 60 h. ( D ) Cells were pre-treated with 40 mg/mL TO for 3 h before transduction with the indicated virus titer for 24 h. After that, the medium was exchanged with fresh medium and incubated for another 48 h. Luminescence was then detected within 1 h. 0.35 mg/mL HMW extract equals to 20 mg/mL TO extract. Transduction control: (−) negative control: bald lentiviral pseudovirion; (+) positive control: firefly luciferase lentivirus; inhibitor positive control: 100 µg/mL anti-hACE2 antibody. ( E ) Pro-inflammatory IL-6 cytokine secretion analysis was done either after 24 h virus transduction together with extract (left), after 24 h + 60 h post-infection with extract (middle) or after 60 h post-infection with extract (right) using multiplexing flow cytometric analysis. Solvent control: distilled water (a.d.). N ≥ 3 independent experiments; * p < 0.05, ** p < 0.01. Significance of difference was calculated relative to the solvent control by one-way ANOVA.

Article Snippet: Anti-hACE2 antibody (Biomol, NSJ-F49433) was used as reference for transduction inhibition assay.

Techniques: Transduction, Inhibition, Infection, Variant Assay, Incubation, Negative Control, Positive Control, Luciferase, Multiplexing