Journal: PLoS ONE

Article Title: Development of ACE2 autoantibodies after SARS-CoV-2 infection

doi: 10.1371/journal.pone.0257016

Figure Lengend Snippet:

Article Snippet: We used the ACE2 inhibitor screening assay kit from AMSBIO (Cambridge, Mass) to measure the ability of plasma or serum from patients in our cohorts to inhibit ACE2 activity.

Techniques:

Journal: PLoS ONE

Article Title: Development of ACE2 autoantibodies after SARS-CoV-2 infection

doi: 10.1371/journal.pone.0257016

Figure Lengend Snippet: There was no statistically significant difference in ACE2 protein between groups. The group labeled + had ACE2 antibodies and the group labeled–did not have ACE2 antibodies. (ns = nonsignificant).

Article Snippet: We used the ACE2 inhibitor screening assay kit from AMSBIO (Cambridge, Mass) to measure the ability of plasma or serum from patients in our cohorts to inhibit ACE2 activity.

Techniques: Labeling

Journal: PLoS ONE

Article Title: Development of ACE2 autoantibodies after SARS-CoV-2 infection

doi: 10.1371/journal.pone.0257016

Figure Lengend Snippet: Activity of the enzyme in plasma was decreased in patients that had an ACE2 antibody present. Bars show mean values and error bars show standard error of the mean (** p<0.01). The group labeled + had ACE2 antibodies and the group labeled–did not have ACE2 antibodies.

Article Snippet: We used the ACE2 inhibitor screening assay kit from AMSBIO (Cambridge, Mass) to measure the ability of plasma or serum from patients in our cohorts to inhibit ACE2 activity.

Techniques: Activity Assay, Labeling

Journal: PLoS ONE

Article Title: Development of ACE2 autoantibodies after SARS-CoV-2 infection

doi: 10.1371/journal.pone.0257016

Figure Lengend Snippet: Plasma from patients with an ACE2 antibody present decreased ACE2 activity. Bars show mean values and error bars show standard error of the mean (*** p<0.05).

Article Snippet: We used the ACE2 inhibitor screening assay kit from AMSBIO (Cambridge, Mass) to measure the ability of plasma or serum from patients in our cohorts to inhibit ACE2 activity.

Techniques: Activity Assay

Journal: PLoS ONE

Article Title: Development of ACE2 autoantibodies after SARS-CoV-2 infection

doi: 10.1371/journal.pone.0257016

Figure Lengend Snippet: The abundance of the ACE2 antibody is plotted against the change in ACE2 activity after addition of plasma.

Article Snippet: We used the ACE2 inhibitor screening assay kit from AMSBIO (Cambridge, Mass) to measure the ability of plasma or serum from patients in our cohorts to inhibit ACE2 activity.

Techniques: Activity Assay

Journal: PLoS ONE

Article Title: Epigallocatechin gallate (EGCG) attenuates severe acute respiratory coronavirus disease 2 (SARS-CoV-2) infection by blocking the interaction of SARS-CoV-2 spike protein receptor-binding domain to human angiotensin-converting enzyme 2

doi: 10.1371/journal.pone.0271112

Figure Lengend Snippet: (A) Schematic of the ELISA-based screening of phytochemical inhibitory activity on ACE2 and SARS-CoV-2 spike RBD binding. Upper scheme: When the compound does not block the ACE2 and spike RBD binding. Lower scheme: When the compound blocks the ACE2 and spike RBD binding. (B) ELISA results of phytochemical inhibition of the ACE2 and spike RBD binding. Low-intensity HRP signals indicate that the compound successfully blocked the ACE2 and spike RBD binding. Values are presented as the mean ± SD. Asterisks indicate the significant difference compared with the DMSO-treated control (** p < 0.01, * p < 0.05). (C) ELISA results of serially diluted EGCG (3.125–100 μM) inhibition of ACE2 and spike RBD binding b. IC50 was calculated using the IC50 calculator ( https://www.aatbio.com/tools/ic50-calculator ).

Article Snippet: We examined the inhibitory effects of SARS-CoV-2 RBD and ACE2 binding by ELISAs using the Spike S1 RBD (SARS-CoV-2): ACE2 Inhibitor Screening Assay Kit (BPS Bioscience, #79931, San Diego, CA, USA) following the manufacturer’s instructions; we tested all compounds at 100 or 20 μM, and we briefly incubated SARS-CoV-2 spike RBD-Fc (50 ng) overnight for 12 h into a nickel-coated 96-well plate.

Techniques: Enzyme-linked Immunosorbent Assay, Activity Assay, Binding Assay, Blocking Assay, Inhibition

Journal: PLoS ONE

Article Title: Epigallocatechin gallate (EGCG) attenuates severe acute respiratory coronavirus disease 2 (SARS-CoV-2) infection by blocking the interaction of SARS-CoV-2 spike protein receptor-binding domain to human angiotensin-converting enzyme 2

doi: 10.1371/journal.pone.0271112

Figure Lengend Snippet: ( A ) Interacting residues determined within a 5-Å region at the interface of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) receptor-binding domain (RBD) and human ACE2 from their crystal structure (6M17). ( B & C ) Molecular docking of human ACE2 and EGCG. ( B ) Three-dimensional representation of EGCG binding with the human ACE2 peptidase domain (left) and magnified image (right). ( C ) Two-dimensional interaction analysis of ACE2 and EGCG, highlighting the ACE2 residues that interact with EGCG and the types and lengths of their bonds. Residues are color coded according to the type of interaction: dark green, conventional hydrogen bonds; light green, van der Waals forces; dark pink, Pi–Pi stacked interactions; and light pink, Pi–alkyl interactions.

Article Snippet: We examined the inhibitory effects of SARS-CoV-2 RBD and ACE2 binding by ELISAs using the Spike S1 RBD (SARS-CoV-2): ACE2 Inhibitor Screening Assay Kit (BPS Bioscience, #79931, San Diego, CA, USA) following the manufacturer’s instructions; we tested all compounds at 100 or 20 μM, and we briefly incubated SARS-CoV-2 spike RBD-Fc (50 ng) overnight for 12 h into a nickel-coated 96-well plate.

Techniques: Binding Assay

Journal: PLoS ONE

Article Title: Epigallocatechin gallate (EGCG) attenuates severe acute respiratory coronavirus disease 2 (SARS-CoV-2) infection by blocking the interaction of SARS-CoV-2 spike protein receptor-binding domain to human angiotensin-converting enzyme 2

doi: 10.1371/journal.pone.0271112

Figure Lengend Snippet: ( A ) Images of 293FT and Caco-2 cells grown on 100-mm plates taken under a phase-contrast microscope. Scale bar: 100 μm. ( B ) Cropped image of western blot performed with ACE2 and Glyceraldehyde 3-phosphate dehydrogenase antibodies. ( C ) Outline of the experimental procedure. Caco-2 cells were treated with or without EGCG for 2 h. After treatment, the cells were washed with PBS thrice, and fresh medium was added. After 22 h of incubation, cell growth was determined. ( D ) Effects of EGCG on Caco-2 cell growth. Cell growth was assessed by the cell proliferation assay and expressed as a percentage of dimethyl sulfoxide (DMSO)-treated control cells. Asterisks indicate the significant difference compared with the DMSO-treated control (** p < 0.01).

Article Snippet: We examined the inhibitory effects of SARS-CoV-2 RBD and ACE2 binding by ELISAs using the Spike S1 RBD (SARS-CoV-2): ACE2 Inhibitor Screening Assay Kit (BPS Bioscience, #79931, San Diego, CA, USA) following the manufacturer’s instructions; we tested all compounds at 100 or 20 μM, and we briefly incubated SARS-CoV-2 spike RBD-Fc (50 ng) overnight for 12 h into a nickel-coated 96-well plate.

Techniques: Microscopy, Western Blot, Incubation, Proliferation Assay

Journal: medRxiv

Article Title: Integrated immune networks in SARS-CoV-2 infected pregnant women reveal differential NK cell and unconventional T cell activation

doi: 10.1101/2021.08.21.21262399

Figure Lengend Snippet: (A) Schematic depicting recruitment of pregnant and non-pregnant women with acute or convalescent COVID-19 or who were not exposed to SARS-CoV-2. (B) Location during acute COVID-19 for pregnant (P) and non-pregnant (Non-P) donors. (C-D) Timeline showing days post disease symptom onset at which blood samples were collected for (C) pregnant and (D) non-pregnant women who had COVID-19. (E) Median days post symptom onset in acute (P n=13, Non-P n=12) and convalescent (P n=15, Non-P n=33) pregnant and non-pregnant donors. Donors with longitudinal sampling are represented for each timepoint collected. (F) Median age of pregnant and non-pregnant healthy (n=21 and 37), acute (n=8 and 9) or convalescent (n=13 and 18) COVID-19 donors. Median and range are shown. (G) Gestational age in weeks for healthy (n=21) and COVID-19 (n=25) pregnant donors. Means and standard deviations are shown. (H) Schematics showing a sub-group of recovered COVID-19 (n=9) or healthy (n=6) pregnant donors from whom matched cord blood and placenta tissue were collected in addition to maternal blood. Median age in years for healthy (n=6) and COVID-19 pregnant donors (n=9). Median and range are shown. (J) Timeline showing the week of gestation at which each pregnant donor became symptomatic with COVID-19 (n=9).

Article Snippet: In addition, SARS-CoV-2 spike trimers (kindly provided by Adam K. Wheatley) and SARS-CoV-2 spike trimers (BPS Bioscience) were also included.

Techniques: Sampling

Journal: medRxiv

Article Title: Integrated immune networks in SARS-CoV-2 infected pregnant women reveal differential NK cell and unconventional T cell activation

doi: 10.1101/2021.08.21.21262399

Figure Lengend Snippet: (Ai) Principal component plots showing pregnant (red, n=13) and non-pregnant (blue, n=11) donors with acute COVID-19. (Aii) Loading plot showing the 14 selected features that cause pregnant and non-pregnant donors to separate along the PC1 axis. Principal component plot of selected features. (Bi) Principal component plots showing pregnant (red, n=8) and non-pregnant (blue, n=10) donors with convalescent COVID-19. (Bii) Loading plot showing the 10 selected features that cause pregnant and non-pregnant donors to separate along the PC1 axis. Principal component plot of selected features. (Ci) Principal component plots showing convalescent pregnant (red, n=8) and cord blood (orange, n=3) from COVID-19 pregnancies. (Cii) Loading plot showing the five selected features that cause pregnant and non-pregnant donors to separate along the PC1 axis. Principal component plot of selected features. IgM and IgA features were excluded from this comparison due to the lack of these isotypes in the cord blood which would masked any antigen-specific findings. SARS2, SARS-CoV-2; SARS1, SARS-CoV-1; S, spike, S1, spike subunit 1 (stalk); S2, spike subunit 2 (head); N, nucleocapsid; RBD, receptor binding domain.

Article Snippet: In addition, SARS-CoV-2 spike trimers (kindly provided by Adam K. Wheatley) and SARS-CoV-2 spike trimers (BPS Bioscience) were also included.

Techniques: Binding Assay

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:

Journal: bioRxiv

Article Title: ACIS, A Novel KepTide™, Binds to ACE-2 Receptor and Inhibits the Infection of SARS-CoV2 Virus in vitro in Primate Kidney Cells: Therapeutic Implications for COVID-19

doi: 10.1101/2020.10.13.337584

Figure Lengend Snippet: Chemiluminometric binding assay of S-glycoprotein RBD and hACE-2(His-tag) with increasing doses of ( A ) KEpTide 1 ( B ) KEpTide 2 (0.5 nM to 100 nM) as directed with manufacturer’s protocol (BPS Bioscience, SARS-CoV2 inhibitor assay kit, Cat # 79936). IC50 value of KEpTide 1 (5.128nM) and KEpTide 2 (24.47 nM) were calculated after logarithmic value of percent inhibition as a function of log doses and then plotted with non-liner regression curve-fit analyses in GraphPad Prism software. Scrambled-peptide did not display any binding. Results are mean ± SD of three independent experiments.

Article Snippet: Donkey anti-mouse Alexa 647-conjugated affinity pure secondary antibody (Cat # 715-605-150) was purchased from Jackson ImmunoResearch Laboratories Inc. Spike RBD (SARS-CoV-2): ACE2 Inhibitor Screening Assay Kit was purchased from BPS Biosciences (Cat # 79931).

Techniques: Binding Assay, Inhibition, Software

Journal: Life

Article Title: 1,2,3-Triazole-Benzofused Molecular Conjugates as Potential Antiviral Agents against SARS-CoV-2 Virus Variants

doi: 10.3390/life12091341

Figure Lengend Snippet: Three-dimensional binding mode of compound 9 (green) at the binding interface between the SARS-CoV-2 S-RBD (red) and human ACE2 (blue).

Article Snippet: SARS-CoV-2 assay kit (SARS-CoV-2 Assay Kit, Biosource, Muskego, WI, USA) and the Omicron variant Spike RBD inhibitor screening kits (B.1.1.529 BA.1, Omicron Variant Assay Kit, BPS Bioscience, San Diego, CA, USA) were used to evaluate the anti-SARS-CoV-2 activity in vitro.

Techniques: Binding Assay

Journal: bioRxiv

Article Title: A novel antibody against the furin cleavage site of SARS-CoV-2 spike protein: effects on proteolytic cleavage and ACE2 binding

doi: 10.1101/2021.02.09.430451

Figure Lengend Snippet: Structure of the SARS-CoV-2 spike protein, including the location of the furin cleavage site at the boundary between the S1 and S2 subunits.

Article Snippet: His-tagged SARS-CoV-2 protein containing the S1/S2 boundary furin site, His-tagged peptide containing the SARS-CoV-2-specific furin motif (EpiGentek), and His-tagged SARS-CoV-2 S1 RBD protein lacking the S1/S2 boundary furin site (EpiGentek) were added at a concentration of 10 ng/well to the Ni-NTA-coated strips and incubated for 1h at 37°C.

Techniques:

Journal: bioRxiv

Article Title: A novel antibody against the furin cleavage site of SARS-CoV-2 spike protein: effects on proteolytic cleavage and ACE2 binding

doi: 10.1101/2021.02.09.430451

Figure Lengend Snippet: fbAB titer of recognizing the antigen containing the SARS-CoV-2-specific furin motif. Antigen concentration = 200 ng/well.

Article Snippet: His-tagged SARS-CoV-2 protein containing the S1/S2 boundary furin site, His-tagged peptide containing the SARS-CoV-2-specific furin motif (EpiGentek), and His-tagged SARS-CoV-2 S1 RBD protein lacking the S1/S2 boundary furin site (EpiGentek) were added at a concentration of 10 ng/well to the Ni-NTA-coated strips and incubated for 1h at 37°C.

Techniques: Concentration Assay

Journal: bioRxiv

Article Title: A novel antibody against the furin cleavage site of SARS-CoV-2 spike protein: effects on proteolytic cleavage and ACE2 binding

doi: 10.1101/2021.02.09.430451

Figure Lengend Snippet: Sensitivity of fbAB recognizing the antigen containing the SARS-CoV-2-specific furin motif.

Article Snippet: His-tagged SARS-CoV-2 protein containing the S1/S2 boundary furin site, His-tagged peptide containing the SARS-CoV-2-specific furin motif (EpiGentek), and His-tagged SARS-CoV-2 S1 RBD protein lacking the S1/S2 boundary furin site (EpiGentek) were added at a concentration of 10 ng/well to the Ni-NTA-coated strips and incubated for 1h at 37°C.

Techniques:

Journal: bioRxiv

Article Title: A novel antibody against the furin cleavage site of SARS-CoV-2 spike protein: effects on proteolytic cleavage and ACE2 binding

doi: 10.1101/2021.02.09.430451

Figure Lengend Snippet: Specificity of fbAB recognizing the SARS-CoV-2 spike protein containing the S1/S2 boundary furin site or a peptide containing the SARS-CoV-2-specific furin motif. A: SARS-CoV-2 protein containing the S1/S2 boundary furin site; B: Peptide containing the SARS-CoV-2-specific furin motif; C: SARS-CoV-2 S1 RBD protein lacking the S1/S2 boundary furin site.

Article Snippet: His-tagged SARS-CoV-2 protein containing the S1/S2 boundary furin site, His-tagged peptide containing the SARS-CoV-2-specific furin motif (EpiGentek), and His-tagged SARS-CoV-2 S1 RBD protein lacking the S1/S2 boundary furin site (EpiGentek) were added at a concentration of 10 ng/well to the Ni-NTA-coated strips and incubated for 1h at 37°C.

Techniques:

Journal: bioRxiv

Article Title: A novel antibody against the furin cleavage site of SARS-CoV-2 spike protein: effects on proteolytic cleavage and ACE2 binding

doi: 10.1101/2021.02.09.430451

Figure Lengend Snippet: Immunoprecipitation of the SARS-CoV-2 protein containing the S1/S2 boundary furin site by fbAB. A: SARS-CoV-2 protein containing the S1/S2 boundary furin site; B: Peptide containing the SARS-CoV-2 specific furin motif.

Article Snippet: His-tagged SARS-CoV-2 protein containing the S1/S2 boundary furin site, His-tagged peptide containing the SARS-CoV-2-specific furin motif (EpiGentek), and His-tagged SARS-CoV-2 S1 RBD protein lacking the S1/S2 boundary furin site (EpiGentek) were added at a concentration of 10 ng/well to the Ni-NTA-coated strips and incubated for 1h at 37°C.

Techniques: Immunoprecipitation

Journal: bioRxiv

Article Title: A novel antibody against the furin cleavage site of SARS-CoV-2 spike protein: effects on proteolytic cleavage and ACE2 binding

doi: 10.1101/2021.02.09.430451

Figure Lengend Snippet: fbAB blockage of SARS-CoV-2 furin motif cleavage by furin. Furin concentration = 2-4 U/well. fbAB and spike protein neutralization antibody (SnAB) concentration = 200 ng/well.

Article Snippet: His-tagged SARS-CoV-2 protein containing the S1/S2 boundary furin site, His-tagged peptide containing the SARS-CoV-2-specific furin motif (EpiGentek), and His-tagged SARS-CoV-2 S1 RBD protein lacking the S1/S2 boundary furin site (EpiGentek) were added at a concentration of 10 ng/well to the Ni-NTA-coated strips and incubated for 1h at 37°C.

Techniques: Concentration Assay, Neutralization

Journal: bioRxiv

Article Title: A novel antibody against the furin cleavage site of SARS-CoV-2 spike protein: effects on proteolytic cleavage and ACE2 binding

doi: 10.1101/2021.02.09.430451

Figure Lengend Snippet: fbAB blockage of SARS-CoV-2 furin motif cleavage by facilitating protease trypsin. Trypsin concentration = 10-20 ng/well. fbAB concentration = 200 ng/well.

Article Snippet: His-tagged SARS-CoV-2 protein containing the S1/S2 boundary furin site, His-tagged peptide containing the SARS-CoV-2-specific furin motif (EpiGentek), and His-tagged SARS-CoV-2 S1 RBD protein lacking the S1/S2 boundary furin site (EpiGentek) were added at a concentration of 10 ng/well to the Ni-NTA-coated strips and incubated for 1h at 37°C.

Techniques: Concentration Assay

Journal: bioRxiv

Article Title: A novel antibody against the furin cleavage site of SARS-CoV-2 spike protein: effects on proteolytic cleavage and ACE2 binding

doi: 10.1101/2021.02.09.430451

Figure Lengend Snippet: fbAB blockage of SARS-CoV-2 furin motif cleavage by human nasal swab sample. Human nasal swab sample was released into 300 µl of furin assay buffer and 20-30 µl of sample solution was used for the assay. fbAB concentration = 200 ng/well.

Article Snippet: His-tagged SARS-CoV-2 protein containing the S1/S2 boundary furin site, His-tagged peptide containing the SARS-CoV-2-specific furin motif (EpiGentek), and His-tagged SARS-CoV-2 S1 RBD protein lacking the S1/S2 boundary furin site (EpiGentek) were added at a concentration of 10 ng/well to the Ni-NTA-coated strips and incubated for 1h at 37°C.

Techniques: Concentration Assay

Journal: bioRxiv

Article Title: A novel antibody against the furin cleavage site of SARS-CoV-2 spike protein: effects on proteolytic cleavage and ACE2 binding

doi: 10.1101/2021.02.09.430451

Figure Lengend Snippet: Reduction of ACE2 binding to SARS-CoV-2 spike protein by fbAB blocking the SARS-CoV-2 furin motif at different concentrations. Coated SARS-CoV-2 spike protein concentration = 50 ng/well. ACE2 concentration = 100 ng/well.

Article Snippet: His-tagged SARS-CoV-2 protein containing the S1/S2 boundary furin site, His-tagged peptide containing the SARS-CoV-2-specific furin motif (EpiGentek), and His-tagged SARS-CoV-2 S1 RBD protein lacking the S1/S2 boundary furin site (EpiGentek) were added at a concentration of 10 ng/well to the Ni-NTA-coated strips and incubated for 1h at 37°C.

Techniques: Binding Assay, Blocking Assay, Protein Concentration, Concentration Assay