recombinant anti ace2 antibody  (Sino Biological)

Journal: Frontiers in Microbiology

Article Title: Broad-spectrum inhibition of SARS-CoV-2 variants by dibutyl phthalate through allosteric disruption of Spike-ACE2 interface

doi: 10.3389/fmicb.2025.1610775

Figure Lengend Snippet: Virtual screening of aromatic compounds effectively inhibiting ACE2-Spike interaction. (A) The heatmap of molecular docking structures of 47 aromatic compounds with different con-formations of Spike and ACE2. The color represents the docking binding energy, with a redder color indicating a more stable binding capacity. The top 10 compounds (indicated in red) were utilized for subsequent experimental detections. (B) Structure and classification of 10 potentially effective compounds. (C) The protein pockets (in gray) where the top 10 compounds (in yellow) bind to different conformations of Spike and ACE2. Spike-RBD-1up/Spike-RBD-3down: RBD is highlighted in solid color; Spike-RBD-2up: The S1 and S2 subunits involved in the pocket are emphasized in solid color; RBD-ACE2: RBD is presented in blue, ACE2 in green, the RBM se-quence in direct contact with ACE2 is in pink, and the interface in direct contact with RBD on ACE2 is in red.

Article Snippet: SARS-CoV-2 Spike trimer protein (40589-V08H4), ACE2 protein (10108-H08H), Recombinant Anti-ACE2 Antibody (10108-R003), SARS-CoV-2 (2019-nCoV) Spike Antibody (Rabbit PAb) (40592-T62) were purchased from sino biological (Beijing, China).

Techniques: Binding Assay

Journal: Frontiers in Microbiology

Article Title: Broad-spectrum inhibition of SARS-CoV-2 variants by dibutyl phthalate through allosteric disruption of Spike-ACE2 interface

doi: 10.3389/fmicb.2025.1610775

Figure Lengend Snippet: SPR Analysis of DBP Binding to ACE2 and SARS-CoV-2 Spike Trimer, and Its Inhibition of Spike-ACE2 Interaction. (A,B) The response curves of DBP (0.0122–3.1250 μM) with ACE2 (40 μg/mL, optimized for DBP-ACE2 binding detection) and S trimer (40 μg/mL, optimized for DBP-S trimer binding detection). (C) Concentration-dependent binding of ACE2 (15.625–250 nM) to S trimer (20 μg/mL, optimized for ACE2-S trimer binding detection). (D) Inhibitory effect of DBP on S trimer-ACE2 interaction. K D : Equilibrium dissociation constant. (E) Proposed mechanism of action of DBP.

Article Snippet: SARS-CoV-2 Spike trimer protein (40589-V08H4), ACE2 protein (10108-H08H), Recombinant Anti-ACE2 Antibody (10108-R003), SARS-CoV-2 (2019-nCoV) Spike Antibody (Rabbit PAb) (40592-T62) were purchased from sino biological (Beijing, China).

Techniques: Binding Assay, Inhibition, Concentration Assay

Journal: Frontiers in Microbiology

Article Title: Broad-spectrum inhibition of SARS-CoV-2 variants by dibutyl phthalate through allosteric disruption of Spike-ACE2 interface

doi: 10.3389/fmicb.2025.1610775

Figure Lengend Snippet: Inhibition of SARS-CoV-2 RBD-ACE2 interaction by DBP and its effect on ACE2 enzymatic activity. (A) Schematic illustration of ELISA assays under three experimental conditions: DBP No Premix, DBP-ACE2 Premix, and DBP-Spike Premix. (B) ELISA showing the inhibitory effect of DBP on the binding of SARS-CoV-2 RBD to ACE2. (C) Bar graph depicting the inhibition rate of ACE2/RBD binding by DBP under varied preincubation conditions. DBP at concentrations of 100 μM (blue) and 200 μM (orange) was evaluated in three conditions: no preincubation (DBP No Premix), preincubation with ACE2 (DBP-ACE2 Premix, 1 h), and preincubation with RBD (DBP-RBD Premix, 1 h). (D) Assessment of DBP’s effect on ACE2 enzymatic activity within a concentration range of 12.5–200 μM. Relative Fluorescence units were reported as mean ± SD. Significant differences were observed in the MLN-4760 group compared to the Neg group (*** P < 0.001), while no statistically significant differences (ns) were detected in the other experimental groups. (E) Molecular docking analysis showing that DBP stably binds at the RBD (Cyan). (F) Structural representation of the ACE2-RBD interface (Red) before DBP binding, showing the formation of 18 hydrogen bonds (Yellow) and one salt bridge (Orange). ACE2 inter-action residues are shown in yellow, RBD residues in salmon. (G) Structural representation of the ACE2-RBD interface (Red) after DBP binding, with only 7 hydrogen bonds remaining.

Article Snippet: SARS-CoV-2 Spike trimer protein (40589-V08H4), ACE2 protein (10108-H08H), Recombinant Anti-ACE2 Antibody (10108-R003), SARS-CoV-2 (2019-nCoV) Spike Antibody (Rabbit PAb) (40592-T62) were purchased from sino biological (Beijing, China).

Techniques: Inhibition, Activity Assay, Enzyme-linked Immunosorbent Assay, Binding Assay, Concentration Assay, Fluorescence, Stable Transfection

Journal: Scientific Reports

Article Title: Impact of in vitro SARS-CoV-2 infection on breast cancer cells

doi: 10.1038/s41598-024-63804-3

Figure Lengend Snippet: SARS-CoV-2 replication in breast cancer cell lines. Western blot and densitometric analysis of ( a ) ACE2 and ( b ) NRP1 protein expression in the MCF7, MDA-MB-231 and HCC1937 breast cancer cell lines. ( c ) ACE2 and NRP1 were silenced by siRNAs in the MCF7, MDA-MB-231 and HCC1937 breast cancer cell lines, which were subsequently infected with SARS-CoV-2 lineage B1. Viral RNA, expressed as copies/100 ng RNA, was quantified in the cytoplasm 6 h p.i. by real-time PCR with primers targeting the viral N1 gene. The data are presented as the means ± SEMs and are representative of one of three independent experiments with similar results. * p < 0.05 by one-way ANOVA followed by Tukey’s multiple comparison test with a single pooled variance. ( d ) MCF7, MDA-MB-231 and HCC1937 breast cancer cells were infected with SARS-CoV-2, and viral RNA was quantified at different time points in the cytoplasm and in the supernatants by real-time PCR. The viral load is expressed as copies/100 ng RNA or copies/mL for the cellular or supernatant RNA, respectively. The data are presented as the means ± SEMs and are representative of one of three independent experiments with similar results. *** p < 0.001 according to two-way ANOVA with multiple comparisons; refers to the comparison between MCF7 cells and MDA-MB-231 cells and the comparison between MCF7 cells and HCC1937 cells.

Article Snippet: The recombinant rabbit monoclonal anti-ACE2 antibody (clone SN0754, dilution 1:1000, Thermo Fisher Scientific, Waltham, MA, USA), rabbit monoclonal anti-Neuropilin-1 (NRP1) antibody (clone D62C6, dilution 1:1000, Cell Signaling Technology Inc., Danvers, MA, USA) and horseradish peroxidase-conjugated mouse monoclonal anti-β-Actin antibody (clone AC-15, dilution 1:30,000, Merck KGaA, Darmstadt, Germany) served as primary antibodies.

Techniques: Western Blot, Expressing, Infection, Real-time Polymerase Chain Reaction, Comparison

Journal: Frontiers in Microbiology

Article Title: Lipid rafts disruption by statins negatively impacts the interaction between SARS-CoV-2 S1 subunit and ACE2 in intestinal epithelial cells

doi: 10.3389/fmicb.2023.1335458

Figure Lengend Snippet: Primary and secondary antibodies.

Article Snippet: Recombinant anti-ACE2 antibody , 0.231 μg/μL , 1:5,000 , Abcam , ab108252.

Techniques: Concentration Assay, Recombinant

Journal: Frontiers in Microbiology

Article Title: Lipid rafts disruption by statins negatively impacts the interaction between SARS-CoV-2 S1 subunit and ACE2 in intestinal epithelial cells

doi: 10.3389/fmicb.2023.1335458

Figure Lengend Snippet: Fluvastatin and simvastatin reduce total ACE2 expression in Caco-2 cells. (A) Caco-2 cells were treated with fluvastatin or simvastatin for 48 h. The cells were lysed, and an equal amount of proteins was analyzed by immunoblotting using anti-ACE2 antibodies. β-actin was used as a loading control. (B) The results obtained in A were normalized to the internal control β-actin. Dunnett’s multiple comparisons test, * p < 0.05, vs. DMSO, S.E.M., n = 3.

Article Snippet: Recombinant anti-ACE2 antibody , 0.231 μg/μL , 1:5,000 , Abcam , ab108252.

Techniques: Expressing, Western Blot

Journal: Frontiers in Microbiology

Article Title: Lipid rafts disruption by statins negatively impacts the interaction between SARS-CoV-2 S1 subunit and ACE2 in intestinal epithelial cells

doi: 10.3389/fmicb.2023.1335458

Figure Lengend Snippet: Fluvastatin and simvastatin affect the sorting of ACE2, DPPIV, and SI to the brush border membrane of Caco-2 cells. (A) Control and treated Caco-2 cells were homogenized 48 h post-treatment. The homogenates were then fractionated into H, P1, P2, and S using the divalent cation (CaCl 2 ) method. Equal amounts of proteins from each fraction were analyzed by immunoblotting with anti-ACE2 antibodies. (B) Results obtained in A were normalized to the respective H, which was used as an internal control. The same procedure as described in A except that (C,D) anti-DPPIV antibodies and (E,F) anti-SI antibodies were used. Tukey’s multiple comparisons test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, vs. DMSO P2, S.E.M., n = 3.

Article Snippet: Recombinant anti-ACE2 antibody , 0.231 μg/μL , 1:5,000 , Abcam , ab108252.

Techniques: Membrane, Western Blot

Journal: Frontiers in Microbiology

Article Title: Lipid rafts disruption by statins negatively impacts the interaction between SARS-CoV-2 S1 subunit and ACE2 in intestinal epithelial cells

doi: 10.3389/fmicb.2023.1335458

Figure Lengend Snippet: Fluvastatin and simvastatin disrupt the association of proteins with lipid rafts (LR). (A) Caco-2 cells were treated with DMSO, fluvastatin, or simvastatin for 48 h. The cells were homogenized, lysed with 1% Lubrol in PBS, and subjected to a discontinuous sucrose gradient. The fractions were then collected, and an equal volume was analyzed by immunoblotting with anti-flottilin-2 antibodies. (B) LR and (C) NLR were normalized to the total fractions. The same procedure described in A, except that (D–F) anti-ACE2 antibodies, (G–I) anti-DPPIV antibodies, and (J–L) anti-SI antibodies were used. Tukey’s multiple comparisons test, * p < 0.05, ** p < 0.01, vs. DMSO, S.E.M., n = 5.

Article Snippet: Recombinant anti-ACE2 antibody , 0.231 μg/μL , 1:5,000 , Abcam , ab108252.

Techniques: Western Blot

Journal: Frontiers in Microbiology

Article Title: Lipid rafts disruption by statins negatively impacts the interaction between SARS-CoV-2 S1 subunit and ACE2 in intestinal epithelial cells

doi: 10.3389/fmicb.2023.1335458

Figure Lengend Snippet: Fluvastatin and simvastatin reduce S1/ACE2 interaction at the cell surface of Caco-2 cells. (A) COS-1 cells were transiently transfected with the S1 subunit and Caco-2 cells were treated with fluvastatin or simvastatin for 48 h. Culture media containing the secreted S1 proteins was collected and added to Caco-2 cells for 2 h at 4°C to allow for S1/ACE2 binding. The cells were then lysed and S1 was captured using Protein A-Sepharose® beads. The samples were analyzed by immunoblotting using anti-ACE2 antibodies. (B) Results in A were analyzed and normalized to the control DMSO sample. Dunnett’s multiple comparisons test, * p < 0.05, ** p < 0.01, vs. DMSO, S.E.M., n = 4.

Article Snippet: Recombinant anti-ACE2 antibody , 0.231 μg/μL , 1:5,000 , Abcam , ab108252.

Techniques: Transfection, Binding Assay, Western Blot

Journal: bioRxiv

Article Title: SARS-CoV-2 infects neurons, astrocytes, choroid plexus epithelial cells and pericytes of the human central nervous system

doi: 10.1101/2023.11.21.568132

Figure Lengend Snippet: Primary human astrocytes, neurons and choroid plexus epithelial cells were incubated with ACE2 neutralizing antibody for 1h (10 μg/mL) before being inoculated with SARS-CoV-2 (Italy_INMI1) at an MOI of 0.1 for 1h. A) 72h post-infection, cells were fixed and immunolabelled for anti-SARS-CoV-2 spike protein, and infected cells enumerated. Data are presented as % neutralization relative to the untreated control. B) Cells were lysed and SARS-CoV-2 nucleocapsid (N1) gene quantified by qRT-PCR. Data are presented as SARS-CoV-2 N1 genome copies/mL (gc/mL). ****p<0.0001, **p<0.01, *p<0.05, ns=not significant. N=3 independent experiments.

Article Snippet: Primary antibodies used for immunofluorescent staining and neutralisation assays were as follows: SARS-CoV-2 Spike mAb anti-rabbit IgG (SinoBiological, Beijing, China); dsRNA K1 mAb anti-mouse IgG2a (Scicons, Susteren, The Netherlands), and recombinant anti-ACE2 neutralising mAb anti-mouse IgG1 (SinoBiological, Beijing, China).

Techniques: Incubation, Infection, Neutralization, Quantitative RT-PCR