Journal: International Journal of Molecular Sciences

Article Title: Mulberry Component Kuwanon C Exerts Potent Therapeutic Efficacy In Vitro against COVID-19 by Blocking the SARS-CoV-2 Spike S1 RBD:ACE2 Receptor Interaction

doi: 10.3390/ijms232012516

Figure Lengend Snippet: The inhibition of the interaction between the spike protein S1 receptor-binding domain (RBD) and the human angiotensin-converting enzyme 2 (ACE2) receptor by kuwanon C (KC). Spike protein coated on a 96-well plate interacted with a preincubated mixture of the ACE2 receptor and ( A ) anti-SARS-CoV-2 spike S1 antibody as the positive control and ( B ) 0, 3.125, 6.25, 12.5, 25, 50, or 100 μM KC. The inhibition of the spike S1 RBD:ACE2 receptor interaction by KC was determined based on chemiluminescence measurements.

Article Snippet: The SARS-CoV-2 spike/ACE2 inhibitor screening assay kit, biotin-labeled recombinant protein ACE2 receptor, and spike protein S1 RBD (BPS Bioscience, San Diego, CA, USA) were purchased for the competitive ELISA assay and BLItz analysis.

Techniques: Inhibition, Binding Assay, Positive Control

Journal: International Journal of Molecular Sciences

Article Title: Mulberry Component Kuwanon C Exerts Potent Therapeutic Efficacy In Vitro against COVID-19 by Blocking the SARS-CoV-2 Spike S1 RBD:ACE2 Receptor Interaction

doi: 10.3390/ijms232012516

Figure Lengend Snippet: The global kinetic analysis of KC binding to biotinylated ( A ) spike S1 RBD- and ( B ) ACE2 receptor-immobilized BLI sensors. The kinetics for the binding of KC to the spike S1 RBD or the ACE2 receptor were measured by the association of 0, 50, 200, and 400 μM of KC in PBS containing 1% DMSO with immobilized spike S1 or ACE2 receptor and the subsequent dissociation in PBS containing 1% DMSO.

Article Snippet: The SARS-CoV-2 spike/ACE2 inhibitor screening assay kit, biotin-labeled recombinant protein ACE2 receptor, and spike protein S1 RBD (BPS Bioscience, San Diego, CA, USA) were purchased for the competitive ELISA assay and BLItz analysis.

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Mulberry Component Kuwanon C Exerts Potent Therapeutic Efficacy In Vitro against COVID-19 by Blocking the SARS-CoV-2 Spike S1 RBD:ACE2 Receptor Interaction

doi: 10.3390/ijms232012516

Figure Lengend Snippet: The binding kinetics of KC to spike S1 RBD and ACE2 receptor.

Article Snippet: The SARS-CoV-2 spike/ACE2 inhibitor screening assay kit, biotin-labeled recombinant protein ACE2 receptor, and spike protein S1 RBD (BPS Bioscience, San Diego, CA, USA) were purchased for the competitive ELISA assay and BLItz analysis.

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Mulberry Component Kuwanon C Exerts Potent Therapeutic Efficacy In Vitro against COVID-19 by Blocking the SARS-CoV-2 Spike S1 RBD:ACE2 Receptor Interaction

doi: 10.3390/ijms232012516

Figure Lengend Snippet: In silico docking simulation between KC and the spike protein/ACE2 receptor. KC was docked onto the SARS-CoV-2 spike protein and ACE2 receptor (PDB code: 6M0J) using AutoDock Vina. The pharmacophore of KC with each target proteins was analyzed using BIOVIA Discovery Studio Visualizer.

Article Snippet: The SARS-CoV-2 spike/ACE2 inhibitor screening assay kit, biotin-labeled recombinant protein ACE2 receptor, and spike protein S1 RBD (BPS Bioscience, San Diego, CA, USA) were purchased for the competitive ELISA assay and BLItz analysis.

Techniques: In Silico

Journal: International Journal of Molecular Sciences

Article Title: Mulberry Component Kuwanon C Exerts Potent Therapeutic Efficacy In Vitro against COVID-19 by Blocking the SARS-CoV-2 Spike S1 RBD:ACE2 Receptor Interaction

doi: 10.3390/ijms232012516

Figure Lengend Snippet: KC inhibits SARS-CoV-2 lentiviral pseudovirus infection in HEK293T cells stably expressing human ACE2 and TMPRSS2. ( A ) The cytotoxic effect of KC in HEK293T cells stably expressing human ACE2 and TMPRSS2 was determined using the MTT assay. HEK293T cells were cultured in 96-well plates (5 × 10 4 cells/well) for 18 h. ( B ) The ACE2 expression level in HEK293T cells was monitored during KC treatment using real-time quantitative PCR analysis. Then, WT or mutant (D614G) SARS-CoV-2 spike pseudovirus (at a final concentration of 1 × 10 4 TU/mL to each well) were mixed with different concentrations of KC (2 and 20 μM) or anti-SARS-CoV-2 antibody, and the mixtures were incubated at 37 °C for 1 h. Then, these mixtures were added to HEK293T cells. ( C , D ) Green fluorescent protein (GFP) expression levels using flow cytometry were assessed at 72 h after viral infection, scale bar = 100 μm. ( E , F ) The inhibitory effect of SARS-CoV-2 spike pseudovirus infection was determined by measuring GFP expression using flow cytometry and measured under a fluorescence microscope. Bar graph (mean ± SEM) statistics were determined from three experimental data sets using one-way ANOVA with Tukey’s post hoc test, *** p < 0.001, compared with the CON (KC-untreated) samples. ### p < 0.001, compared with the cell-only sample.

Article Snippet: The SARS-CoV-2 spike/ACE2 inhibitor screening assay kit, biotin-labeled recombinant protein ACE2 receptor, and spike protein S1 RBD (BPS Bioscience, San Diego, CA, USA) were purchased for the competitive ELISA assay and BLItz analysis.

Techniques: Infection, Stable Transfection, Expressing, MTT Assay, Cell Culture, Real-time Polymerase Chain Reaction, Mutagenesis, Concentration Assay, Incubation, Flow Cytometry, Fluorescence, Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Mulberry Component Kuwanon C Exerts Potent Therapeutic Efficacy In Vitro against COVID-19 by Blocking the SARS-CoV-2 Spike S1 RBD:ACE2 Receptor Interaction

doi: 10.3390/ijms232012516

Figure Lengend Snippet: KC suppresses the infection of a clinical isolate of SARS-CoV-2 alpha strain (βCoV/Korea/KCDC03/2020) in Vero cells. Vero cells were cultured on 384-well plates (1.2 × 10 4 cells/well) for 24 h. Then, Vero cells were infected with SARS-CoV-2 (MOI 0.0125) immediately after being treated with serially diluted KC and incubated at 37 °C for 24 h. The cells were then stained using anti-SARS-CoV-2 nucleocapsid (N) primary antibody, Alexa Fluor 488-conjugated goat antirabbit IgG secondary antibody, and Hoechst 33342.

Article Snippet: The SARS-CoV-2 spike/ACE2 inhibitor screening assay kit, biotin-labeled recombinant protein ACE2 receptor, and spike protein S1 RBD (BPS Bioscience, San Diego, CA, USA) were purchased for the competitive ELISA assay and BLItz analysis.

Techniques: Infection, Cell Culture, Incubation, Staining

Journal: International Journal of Molecular Sciences

Article Title: Mulberry Component Kuwanon C Exerts Potent Therapeutic Efficacy In Vitro against COVID-19 by Blocking the SARS-CoV-2 Spike S1 RBD:ACE2 Receptor Interaction

doi: 10.3390/ijms232012516

Figure Lengend Snippet: Schematic of the blockade of the SARS-CoV-2 spike S1 RBD:ACE2 receptor interaction by KC.

Article Snippet: The SARS-CoV-2 spike/ACE2 inhibitor screening assay kit, biotin-labeled recombinant protein ACE2 receptor, and spike protein S1 RBD (BPS Bioscience, San Diego, CA, USA) were purchased for the competitive ELISA assay and BLItz analysis.

Techniques:

Journal: Scientific Reports

Article Title: A bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels

doi: 10.1038/s41598-021-97330-3

Figure Lengend Snippet: Validation of Lumit SARS-CoV-2 Spike RBD:ACE2 Immunoassay. ( a ) Assay linearity of Lumit SARS-CoV-2 Spike RBD:ACE2 Immunoassay. SARS-CoV-2 Spike RBD:ACE2 titration was performed in 1× Buffer. ( b ) Spike RBD:ACE2-mediated antibody proximity leads to NanoLuc complementation and bioluminescent signal. The absence of either assay component results in loss of signal. Bioluminescence is dependent on the RBD:ACE2 interaction as evidenced by inhibition with anti-Spike RBD antibody. ( c ) SARS-CoV-2 Spike RBD:ACE2 detection is not affected by the orientation of NanoLuc reporter fragment in secondary antibodies. Set 1: Lumit Anti-Mouse Ab-LgBiT/Lumit Anti-Rabbit Ab-SmBiT. Set 2: Lumit Anti-Mouse Ab-SmBiT/Lumit Anti-Rabbit Ab-LgBiT. ( d ) RBD:ACE2 interaction is inhibited by soluble ACE2-His. As control, ACE2-His dilutions were incubated with a luminescence-generating reaction that contained either 1.5 nM RBD-rabbit Fc and Lumit Anti-Rabbit (SmBit/LgBit) antibodies, or 1.5 nM ACE2-mouse Fc and Lumit Anti-Mouse (SmBit/LgBit) antibodies in 1× Lumit immunoassay reaction buffer, and assay was performed as described above. Results are presented as means ± S.E.M. (n = 3 technical replicates, the data are representative of two or more experiments).

Article Snippet: Lumit SARS-CoV-2 Spike RBD:ACE2 immunoassay components were from Promega (Madison, USA) and they consist of 0.5 μM SARS-CoV-2 RBD-rabbit Fc (RBD-Fc), 0.5 μM human ACE2-mouse Fc (ACE2-Fc), Lumit Detection Substrate C, 10X Lumit Immunoassay Buffer C, Lumit anti-rabbit Ab-SmBiT and Lumit anti-mouse Ab-LgBiT.

Techniques: Titration, Inhibition, Incubation

Journal: Scientific Reports

Article Title: A bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels

doi: 10.1038/s41598-021-97330-3

Figure Lengend Snippet: Discovery and characterization of Spike RBD:ACE2 inhibitory molecules. ( a ) Antibody screening. Anti-SARS-CoV-1 and 2 Spike antibodies were tested at 55 nM. Anti-SARS-CoV-2 Nucleocapsid (NP) antibody (#16) was used as a control for specificity and the absence of antibody (Ctrl) was used for 100% activity. ( b ) Antibody dose response experiments. Monoclonal antibodies were serially diluted and incubated with SARS-CoV-2 Spike RBD-rabbit Fc for 30 min prior to the addition of hACE2-mouse Fc and Lumit antibodies. ( c ) Inhibitory peptides were tested at 1 µM for their ability to disrupt the RBD:ACE2 interaction and the absence of peptide (Ctrl) was used for 100% activity. ( d ) Peptides were serially diluted and incubated with SARS-CoV-2 Spike RBD-rabbit Fc for 30 min prior to the addition of hACE2-mouse Fc and Lumit antibodies. The antibodies and peptides in b and d were tested with either RBD-Fc and Lumit Anti-Rabbit antibodies, or with hACE2-Fc and Lumit Anti-mouse antibodies and showed no interference with NanoBit luciferase activity. Results are presented as means ± S.E.M. (n = 3 technical replicates, the data are representative of two or more experiments).

Article Snippet: Lumit SARS-CoV-2 Spike RBD:ACE2 immunoassay components were from Promega (Madison, USA) and they consist of 0.5 μM SARS-CoV-2 RBD-rabbit Fc (RBD-Fc), 0.5 μM human ACE2-mouse Fc (ACE2-Fc), Lumit Detection Substrate C, 10X Lumit Immunoassay Buffer C, Lumit anti-rabbit Ab-SmBiT and Lumit anti-mouse Ab-LgBiT.

Techniques: Activity Assay, Incubation, Luciferase

Journal: Scientific Reports

Article Title: A bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels

doi: 10.1038/s41598-021-97330-3

Figure Lengend Snippet: Effect of RBD mutations on Spike RBD:ACE2 binding. ( a ) Schematic diagram showing SARS-CoV-2 Spike RBD mutations evaluated in this study. ( b ) Rationale of the competition assay used for evaluation of RBD mutations. RBD-His mutants were evaluated in dose response experiments, and IC 50 values generated were compared with wild type RBD-His. ( c , d ) RBD-His WT and mutants were serially diluted and incubated with hACE2-mouse Fc for 30 min prior to the addition of RBD-rabbit Fc and Lumit antibodies. ( e ) Antibody screening using wild type RBD-Fc and variants containing the E484K, Y453F or N501Y mutations. ( f – h ) Antibody titrations with RBD-Fc WT and mutants. Results are presented as means ± S.E.M. (n = 3 technical replicates, the data are representative of two or more experiments).

Article Snippet: Lumit SARS-CoV-2 Spike RBD:ACE2 immunoassay components were from Promega (Madison, USA) and they consist of 0.5 μM SARS-CoV-2 RBD-rabbit Fc (RBD-Fc), 0.5 μM human ACE2-mouse Fc (ACE2-Fc), Lumit Detection Substrate C, 10X Lumit Immunoassay Buffer C, Lumit anti-rabbit Ab-SmBiT and Lumit anti-mouse Ab-LgBiT.

Techniques: Binding Assay, Competitive Binding Assay, Generated, Incubation

Journal: Scientific Reports

Article Title: A bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels

doi: 10.1038/s41598-021-97330-3

Figure Lengend Snippet: Schematic representation of the Lumit SARS-CoV-2 Spike RBD:ACE2 Immunoassay and comparison with PRNT assay. ( a ) Lumit SARS-CoV-2 Spike RBD:ACE2 Immunoassay detects the interaction between the rabbit Fc-tagged RBD fragment of SARS-CoV-2 Spike protein and the soluble mouse Fc-tagged ACE2 receptor through binding of LgBit/SmBit-conjugated Lumit secondary antibodies. Lumit secondary antibody recognition of RBD:ACE2 interaction leads to NanoLuc fragments complementation and bioluminescent signal. Lumit SARS-Cov-2 Spike RBD:ACE2 Immunoassay protocol. Sample (i.e.: antibody, inhibitor, serum, plasma, etc.) is co-incubated with Spike RBD-rabbit Fc for 30 min at room temperature. hACE2-mouse Fc and Lumit secondary antibodies are added to the samples and the assay reactions are incubated at RT for 60 min. Bioluminescence is recorded in a luminometer after the addition of Detection Reagent and incubation for 30 min at room temperature. ( b ) Plaque Reduction Neutralization Test. Serially diluted patient samples are incubated with viable SARS-CoV-2 viral particles before dispensing onto VeroE6 cells. Formation of plaques due to cytopathic effect are inhibited if neutralizing antibodies are present, and the titer is determined by the serum dilution required to inhibit plaque formation by 90% or 50% (i.e.: PRNT90 or PRNT50, respectively).

Article Snippet: Lumit SARS-CoV-2 Spike RBD:ACE2 immunoassay components were from Promega (Madison, USA) and they consist of 0.5 μM SARS-CoV-2 RBD-rabbit Fc (RBD-Fc), 0.5 μM human ACE2-mouse Fc (ACE2-Fc), Lumit Detection Substrate C, 10X Lumit Immunoassay Buffer C, Lumit anti-rabbit Ab-SmBiT and Lumit anti-mouse Ab-LgBiT.

Techniques: Plaque Reduction Neutralization Test, Binding Assay, Incubation

Journal: Scientific Reports

Article Title: A bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels

doi: 10.1038/s41598-021-97330-3

Figure Lengend Snippet: Testing patient derived samples with Lumit SARS-CoV-2 Spike RBD:ACE2 Immunoassay. ( a ) Patient-derived serum samples from 41 COVID-19 positive and 43 negative pre-pandemic samples were tested using Lumit SARS-CoV-2 Spike RBD:ACE2 Immunoassay at 1/20 (5%) final serum dilution. Serum samples were pre-incubated with RBD-Fc for 30 min prior to the addition of the other assay components. ( b ) COVID-19 positive (n = 116) and negative pre-pandemic (n = 202) plasma samples were tested at 1/20 final plasma dilution. A cutoff value of 30% was selected based on the distribution of COVID-19 positive and negative pre-pandemic samples and the assay improved specificity. The horizontal lines indicate the mean values, and the dotted lines represent the cutoff at 30% inhibition. The P values presented in ( a ) and ( b ) were calculated from unpaired two-tailed Student’s t tests. ( c ) Assay robustness. COVID-19 positive (n = 58) and negative pre-pandemic (n = 101) plasma samples were tested on two different dates. ( d ) COVID-19 positive plasma samples with varying percent neutralization values were titrated to determine the NC 50 values. ( e ) Correlation analysis of Lumit Spike RBD:ACE2 Immunoassay with an in vitro surrogate viral neutralization assay (sVNT). Serially diluted plasma samples with percent neutralization values ranging from 50 to 90% at 1/20 final plasma dilution were tested with both Lumit and the sVNT (c-Pass), and NC 50 values were determined and compared. The data presented are the log of the IC 50 values for sVNT and Lumit and are the mean from two independent experiments. Correlation and linear regression analyses in c and e were performed in GraphPad Prism using Pearson’s correlation coefficients. Statistical significance was calculated using the two-tailed test.

Article Snippet: Lumit SARS-CoV-2 Spike RBD:ACE2 immunoassay components were from Promega (Madison, USA) and they consist of 0.5 μM SARS-CoV-2 RBD-rabbit Fc (RBD-Fc), 0.5 μM human ACE2-mouse Fc (ACE2-Fc), Lumit Detection Substrate C, 10X Lumit Immunoassay Buffer C, Lumit anti-rabbit Ab-SmBiT and Lumit anti-mouse Ab-LgBiT.

Techniques: Derivative Assay, Incubation, Inhibition, Two Tailed Test, Neutralization, In Vitro

Journal: Scientific Reports

Article Title: A bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels

doi: 10.1038/s41598-021-97330-3

Figure Lengend Snippet: Evaluation of patient-derived samples from a Brazilian cohort. ( a ) COVID-19 positive (n = 87) and negative pre-pandemic (n = 55) samples were tested using the Lumit SARS-CoV-2 Spike RBD:ACE2 Immunoassay at 1/20 (5%) final serum dilution. The sensitivity and specificity values are 59% and 96%, respectively. The horizontal lines indicate the mean values, and the dotted line represents the cutoff at 30% inhibition. The P value presented was calculated from unpaired two-tailed Student’s t tests. ( b ) Lumit Detection of neutralizing antibodies in PCR-COVID-19 positive samples over time and comparison with results generated by an ELISA-based serological assay ( c ) Percent correlation between Lumit and ELISA-based serological assay data, which targets the trimeric Spike protein ( d ) Antibodies that disrupt the Spike RBD:ACE2 interaction were monitored for at least 90 days after the initial symptom onset in serum of 25 patients in the Brazilian patient cohort. Results are representative of two independent experiments. ( e ) Time-course of neutralizing anti-Spike RBD antibodies in immunized individuals. Individuals immunized with CoronaVac vaccine (n = 28) were tested using Lumit SARS-CoV-2 Spike RBD:ACE2 Immunoassay. Serum samples collected before and throughout the vaccination period were pre-incubated with RBD-rabbit Fc for 30 min prior to the addition of the other assay components. The number of samples tested positive for the presence of neutralizing antibodies increase after the second dose (i.e.: day zero). First and second dose days are represented in vertical lines.

Article Snippet: Lumit SARS-CoV-2 Spike RBD:ACE2 immunoassay components were from Promega (Madison, USA) and they consist of 0.5 μM SARS-CoV-2 RBD-rabbit Fc (RBD-Fc), 0.5 μM human ACE2-mouse Fc (ACE2-Fc), Lumit Detection Substrate C, 10X Lumit Immunoassay Buffer C, Lumit anti-rabbit Ab-SmBiT and Lumit anti-mouse Ab-LgBiT.

Techniques: Derivative Assay, Inhibition, Two Tailed Test, Generated, Enzyme-linked Immunosorbent Assay, Serologic Assay, Incubation

Journal: Molecular Informatics

Article Title: Screening of Clinically Approved and Investigation Drugs as Potential Inhibitors of SARS‐CoV‐2: A Combined in silico and in vitro Study

doi: 10.1002/minf.202100062

Figure Lengend Snippet: Box and Whisker plot representation of MM/GBSA scores of selected hit compounds at the SARS‐CoV‐2 Main Protease target. In the plots, central line in box represents median, bottom line of box is first quartile (25 %), top line of box is third quartile (75 %), bottom of whiskers is first quarter minus 1.5× interquartile range, top of whiskers is third quarter plus 1.5× interquartile range, and dots are outliers.

Article Snippet: Two different inhibitor screening assay kits against Spike protein were used as ACE2:SARS‐CoV‐2 Spike S1 (#79945) and SARS‐CoV‐2 Spike:ACE2 (Cat. #79931), (BPS Bioscience).

Techniques: Whisker Assay

Journal: Molecular Informatics

Article Title: Screening of Clinically Approved and Investigation Drugs as Potential Inhibitors of SARS‐CoV‐2: A Combined in silico and in vitro Study

doi: 10.1002/minf.202100062

Figure Lengend Snippet: Box and Whisker plot representation of MM/GBSA scores of selected hit compounds at the SARS‐CoV‐2 Spike/ACE2 target. In the plots, central line in box represrents median, bottom line of box is first quartile (25 %), top line of box is third quartile (75 %), bottom of whiskers is first quarter minus 1.5× interquartile range, top of whiskers is third quarter plus 1.5× interquartile range, and dots are outliers.

Article Snippet: Two different inhibitor screening assay kits against Spike protein were used as ACE2:SARS‐CoV‐2 Spike S1 (#79945) and SARS‐CoV‐2 Spike:ACE2 (Cat. #79931), (BPS Bioscience).

Techniques: Whisker Assay

Journal: Molecular Informatics

Article Title: Screening of Clinically Approved and Investigation Drugs as Potential Inhibitors of SARS‐CoV‐2: A Combined in silico and in vitro Study

doi: 10.1002/minf.202100062

Figure Lengend Snippet: Change in SARS‐CoV‐2 Spike/ACE2 binding with different inhibitor concentrations. The compound was tested at least with four replicates. Mean and standard deviation results were provided at the figure.

Article Snippet: Two different inhibitor screening assay kits against Spike protein were used as ACE2:SARS‐CoV‐2 Spike S1 (#79945) and SARS‐CoV‐2 Spike:ACE2 (Cat. #79931), (BPS Bioscience).

Techniques: Binding Assay, Standard Deviation

Journal: The Lancet Regional Health: Western Pacific

Article Title: Immunogenicity and safety of heterologous immunisation with Ad5-nCOV in healthy adults aged 60 years and older primed with an inactivated SARS-CoV-2 vaccine (CoronaVac): a phase 4, randomised, observer-blind, non-inferiority trial

doi: 10.1016/j.lanwpc.2023.100829

Figure Lengend Snippet: Serum neutralising activities against SARS-CoV2 wild-type and variants of concern (beta, delta and omicron) before and after boosting, measured by ACE2-binding assay. Neutralising capacity measured by ACE2-binding assay in serum of participants who had received heterologous Convidecia and homologous CoronaVac immunisation at day 0 (a) and day 14 (b). Inhibition of ACE2 binding was measured against the wide-type, beta, delta, and omicron Spike proteins. Horizontal bars show the median, and error bars indicate interquartile range (IQR). Group A, primed with two doses of CoronaVac and given one dose of Convidecia (n = 50); Group B, primed with two doses of CoronaVac and given one dose of CoronaVac (n = 50); ACE2, angiotensin-converting enzyme 2.

Article Snippet: Additionally, we used the Spike- ACE2 binding inhibition assay (Meso Scale Discovery (MSD) platform) to test the functional neuralization capacity in samples from the first 100 participants in the three-dose regimen (50 each from the homologous and heterologous boost cohorts) as a post-hoc measurement for cross-reaction of beta (B.1.351), delta (B.1.617.2), and omicron subvariants (BA.2, BA.3, BA.4 and BA.5).

Techniques: Binding Assay, Inhibition

Journal: bioRxiv

Article Title: Bcr-Abl tyrosine kinase inhibitor imatinib as a potential drug for COVID-19

doi: 10.1101/2020.06.18.158196

Figure Lengend Snippet: A] The association and dissociation curves obtained by BLI reflecting the binding of imatinib to immobilized SARS-CoV-2 RBD protein. Data fitted using the 1:1 binding model are shown in black. B] In vitro colorimetric assay for evaluation of spike RBD (SARS-CoV-2):ACE2 interaction inhibition in the presence of imatinib. Here, 5-fold dilution of imatinib were done in triplicate. SARS-CoV-2 spike RBD (1 μg/ml) and ACE2 (1 μg/ml) were incubated for 1 hour at room temperature with slow shaking in the presence of various imatinib concentrations. Streptavidin HRP (1:1,000) was added to the reaction mixture. Colorimetric substrate was added to initiate the chromogenic reaction, and 2 minutes were allowed for color development. The reaction was terminated with the addition of 1N HCl and absorbance was measured at 450 nm. Positive control (no inhibitor) was assumed to represent 0% inhibition. Values obtained from test wells (with imatinib) compared to the positive control showed 0% inhibition of RBD:ACE2 interaction, indicating that imatinib does not inhibit spike fusion by direct inhibition.

Article Snippet: The ability of imatinib to inhibit the interaction of spike RBD:ACE2 proteins was evaluated by using the Spike RBD (SARS-CoV-2): ACE2 Inhibitor Screening Colorimetric Assay Kit (BPS Bioscience).

Techniques: Binding Assay, In Vitro, Colorimetric Assay, Inhibition, Incubation, Positive Control

Journal: Bioorganic Chemistry

Article Title: Synthesis of novel calcium channel blockers with ACE2 inhibition and dual antihypertensive/anti-inflammatory effects: A possible therapeutic tool for COVID-19

doi: 10.1016/j.bioorg.2021.105272

Figure Lengend Snippet: Effect of synthetized derivatives on ACE2:SARS-CoV-2 Spike (RBD) inhibition.

Article Snippet: In order to measure Angiotensin-Converting Enzyme 2 (ACE2) receptor inhibition activity, the ACE2:SARS-CoV-2 Spike (RBD) Inhibitor Screening Colorimetric Assay Kit # 78031 (Bps Bioscience, Cornerstone Court W, Ste B San Diego, CA 92121) was used.

Techniques: Inhibition

Journal: Bioorganic Chemistry

Article Title: Synthesis of novel calcium channel blockers with ACE2 inhibition and dual antihypertensive/anti-inflammatory effects: A possible therapeutic tool for COVID-19

doi: 10.1016/j.bioorg.2021.105272

Figure Lengend Snippet: SAR for the most potent compounds. DHPMs: Dihydropyrimidines, CCBs: calcium channel blockers, ACE2: Angiotensin-converting enzyme 2, THP-1: Human monocytic cell line, IL-6: Interleukine-6, LPS: Lipopolysaccharide, CRP: C-Reactive Protein.

Article Snippet: In order to measure Angiotensin-Converting Enzyme 2 (ACE2) receptor inhibition activity, the ACE2:SARS-CoV-2 Spike (RBD) Inhibitor Screening Colorimetric Assay Kit # 78031 (Bps Bioscience, Cornerstone Court W, Ste B San Diego, CA 92121) was used.

Techniques: