s1 subunit, receptor-binding domain  (InterPro Inc)

Journal: International Archives of Allergy and Immunology

Article Title: Quantitative Measurement of IgG to Severe Acute Respiratory Syndrome Coronavirus-2 Proteins Using ImmunoCAP

doi: 10.1159/000514203

Figure Lengend Snippet: a Dilution curves for IgG to SARS-CoV-2 spike-RBD and nucleocapsid (5-fold dilutions, n = 5). b Comparison of results of IgG to SARS-CoV-2 nucleocapsid using ImmunoCAP and Abbott assay (n = 10). c Concentration response curve using the anti-spike glycoprotein S1 monoclonal antibody CR3022 (2-fold dilutions carried out in triplicate ±SD). d IgG to SARS-CoV-2 spike-RBD and nucleocapsid in COVID-19 patients (n = 51) and controls (n = 109). SARS-CoV-2, severe acute respiratory syndrome coronavirus-2; COVID-19, coronavirus disease 2019; RBD, receptor-binding domain.

Article Snippet: Reagents Recombinant SARS-CoV-2 S1 receptor-binding domain (RBD) subunit and recombinant SARS-CoV-2 nucleocapsid protein which had been expressed in HEK293 cells were purchased from RayBiotech (Peachtree Corners, GA, USA).

Techniques: Concentration Assay, Binding Assay

Journal: International Archives of Allergy and Immunology

Article Title: Quantitative Measurement of IgG to Severe Acute Respiratory Syndrome Coronavirus-2 Proteins Using ImmunoCAP

doi: 10.1159/000514203

Figure Lengend Snippet: Longitudinal sampling of IgG to SARS-CoV-2 spike-RBD (a) and nucleocapsid with ImmunoCAP (b) in 17 patients hospitalized with COVID-19. c IgG to spike-RBD and nucleocapsid in individual patients. All samples after day 40 were obtained from a post-hospital follow-up clinic. SARS-CoV-2, severe acute respiratory syndrome coronavirus-2; COVID-19, coronavirus disease 2019; RBD, receptor-binding domain.

Article Snippet: Reagents Recombinant SARS-CoV-2 S1 receptor-binding domain (RBD) subunit and recombinant SARS-CoV-2 nucleocapsid protein which had been expressed in HEK293 cells were purchased from RayBiotech (Peachtree Corners, GA, USA).

Techniques: Sampling, Binding Assay

Journal: International Archives of Allergy and Immunology

Article Title: Quantitative Measurement of IgG to Severe Acute Respiratory Syndrome Coronavirus-2 Proteins Using ImmunoCAP

doi: 10.1159/000514203

Figure Lengend Snippet: a Levels of IgG to SARS-CoV-2 spike-RBD, non-SARS coronaviruses and reference antigens with ImmunoCAP, and also total IgG, among controls (n = 29) and COVID-19 patients sampled in the subacute (n = 15, denoted in red) or convalescent (n = 36, denoted in orange) phase. b IgG levels expressed in relation to total IgG. c Relationship between IgG to SARS-CoV-2 at the subacute time point and IgG to non-SARS HCoV. SARS-CoV-2, severe acute respiratory syndrome coronavirus-2; COVID-19, coronavirus disease 2019; RBD, receptor-binding domain.

Article Snippet: Reagents Recombinant SARS-CoV-2 S1 receptor-binding domain (RBD) subunit and recombinant SARS-CoV-2 nucleocapsid protein which had been expressed in HEK293 cells were purchased from RayBiotech (Peachtree Corners, GA, USA).

Techniques: Binding Assay

Journal: bioRxiv

Article Title: The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in vitro models of the human blood–brain barrier

doi: 10.1101/2020.06.15.150912

Figure Lengend Snippet: Paraffin-embedded brain tissue was sectioned at 5 microns and immunostained for ACE2. (A) shows frontal cortex from a patient with no abnormal neuropathology (control) and (B) from a patient diagnosed with mixed dementia. The representative images at 40x objective magnification, shows the ACE2 expression in blue (Vector Blue). The images were selected to demonstrate immunopositive ACE2 on a range of vessel calibers. Arrow heads indicates the vascular presentation of ACE2 expression compared to ACE2 expression in parenchymal cells. Scalebars = 25 microns. (C) Western blots of hBMVEC cell lysates probed with ACE2 antibodies after cells were exposed to 10nM of SARS-CoV-2 spike proteins S1-RBD and S2. (D) Bar graph of densitometry quantification (average ± SEM) for the expression of ACE2 normalized to β-Actin is shown. Statistical significance differences, *P < 0.05, compared with the untreated control (n=3 donors performed in triplicate).

Article Snippet: Spike protein S1 subunit (RayBiotech, Cat No 230-01101), Spike protein S1 subunit Receptor Binding Domain sequence (RBD) (RayBiotech, Cat No 230-01102), and Spike protein S2 subunit (RayBiotech, Cat No 230-01103) were used for in vitro experiments.

Techniques: Expressing, Plasmid Preparation, Western Blot

Journal: bioRxiv

Article Title: The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in vitro models of the human blood–brain barrier

doi: 10.1101/2020.06.15.150912

Figure Lengend Snippet: hBMVECs were treated with 1nM and 10nM of the SARS-CoV2 spike proteins (S1 full, S1-RBD, and S2 full) for 48 hrs (A) and 72 hrs (B). Cell viability was determined using the Live/Dead Cytotoxicity assay. Calcein positive (green) indicates live cells while ethidium homodimer-1 (EthD-1, red) indicates dead cells. Saponin was used a positive control. Data obtained from two different donors, each performed in 6 replicates. mean ± SD *p<0.05.

Article Snippet: Spike protein S1 subunit (RayBiotech, Cat No 230-01101), Spike protein S1 subunit Receptor Binding Domain sequence (RBD) (RayBiotech, Cat No 230-01102), and Spike protein S2 subunit (RayBiotech, Cat No 230-01103) were used for in vitro experiments.

Techniques: Cytotoxicity Assay, Positive Control

Journal: bioRxiv

Article Title: The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in vitro models of the human blood–brain barrier

doi: 10.1101/2020.06.15.150912

Figure Lengend Snippet: (A-C) Barrier electrical resistance was modelled based on continuous cell-substrate impedance readings recorded at 6 frequencies (400Hz – 48kHz) every 6 min for the duration of the time shown. Endothelial monolayers were treated with 0.1nM, 1nM or 10nM of S1 full, S1 RBD, S2 full or left untreated to serve as a baseline. Treatments were initiated at 0 timepoint. Experiment was performed in quadruplicates and repeated three times using primary cells obtained from three different donors. Each data point is represented as the percentage of the mean value ± SEM. D. Endothelial monolayers were treated with 100ng/mL TNF-a, 10nM of S1 full, S1 RBD, S2 full or left untreated to serve as a baseline. Barrier permeability to small molecular tracer was modelled using FITC-conjugated DEAE-dextran (3 kDa). Experiment was performed in quadruplicates and repeated three times using primary cells obtained from three different donors. Each data point is represented as the percentage of the mean value ± SD. p-values were computed using one-way ANOVA and Turkey post-hoc test.

Article Snippet: Spike protein S1 subunit (RayBiotech, Cat No 230-01101), Spike protein S1 subunit Receptor Binding Domain sequence (RBD) (RayBiotech, Cat No 230-01102), and Spike protein S2 subunit (RayBiotech, Cat No 230-01103) were used for in vitro experiments.

Techniques: Permeability

Journal: bioRxiv

Article Title: The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in vitro models of the human blood–brain barrier

doi: 10.1101/2020.06.15.150912

Figure Lengend Snippet: Human brain microvascular endothelial cells (hBMVECs) were treated with 10nM of S1 full, S1 RBD, S2 full or 100ng/mL of TNF-α for 4 and 24hr. Cells were stained for ICAM-1 and VCAM-1 expression and analyzed using a FACS Canto II flow cytometer. Shown are representative histograms for ICAM-1 expression in response to S1 full (A), S1-RBD (B), S2 full (C), TNF-α (D) and the bar graph quantification of the Mean Fluorescent Intensity (MFI) (E). Representative histogram for VCAM-1 expression in response to S1 full (F), S1 RBD (G), S2 full (H), TNF-α (J) and the bar graph quantification of MFI (J). (n=3, *p<0.05).

Article Snippet: Spike protein S1 subunit (RayBiotech, Cat No 230-01101), Spike protein S1 subunit Receptor Binding Domain sequence (RBD) (RayBiotech, Cat No 230-01102), and Spike protein S2 subunit (RayBiotech, Cat No 230-01103) were used for in vitro experiments.

Techniques: Staining, Expressing, Flow Cytometry

Journal: bioRxiv

Article Title: The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in vitro models of the human blood–brain barrier

doi: 10.1101/2020.06.15.150912

Figure Lengend Snippet: Confluent BMVECs monolayers were incubated with 10nM of S1 full, S1-RBD, S2 full for time indicated or left untreated to serve as a control. mRNA expression of cytokine genes after stimulation with the SARS-CoV-2 spike protein S1, S1-RBD and S2 subunits (all at 10nM) are shown for either 4 hrs and 24 hrs. Taqman gene expression assays were performed in quadruplicates and repeated three times using primary cells obtained from three different donors. Target cytokine genes analyzed included: IL1β, IL6, CCL5, CXCL10 at 4hrs (A) and 24 hrs (B). Gene expression analysis for MMP2, MMP3, MMP9, MMP12 and the MMP inhibitor TIMP1 are shown for 4 hrs (C) and 24 hrs (D) respectively. Each bar represents fold-change ± SEM. Data sets were separately analyzed using one-way ANOVA and p-values were computed using Turkey post-hoc test.

Article Snippet: Spike protein S1 subunit (RayBiotech, Cat No 230-01101), Spike protein S1 subunit Receptor Binding Domain sequence (RBD) (RayBiotech, Cat No 230-01102), and Spike protein S2 subunit (RayBiotech, Cat No 230-01103) were used for in vitro experiments.

Techniques: Incubation, Expressing