Journal: bioRxiv

Article Title: A protective broadly cross-reactive human antibody defines a conserved site of vulnerability on beta-coronavirus spikes

doi: 10.1101/2021.03.30.437769

Figure Lengend Snippet: A . Neutralization of SARS-CoV-2 and SARS-CoV-1 by CC40.8 mAb isolated from a COVID-19 donor. B . CELISA binding of CC40.8 mAb with β-HCoV spikes expressed on 293T cells. Binding to HCoV spikes is recorded as % positive cells using a flow cytometry method. CC40.8 mAb shows cross-reactive binding with 4 out of 5 human β-HCoV spikes. BioLayer Interferometry (BLI) binding of CC40.8 mAb with human β-HCoV soluble S proteins. Binding constants (KDs) for each Ab-antigen interaction are indicated. C . Epitope mapping of CC40.8 with HCoV-HKU1 S2 subunit overlapping peptides. A series of HCoV-HKU1 S2 overlapping biotinylated peptides (15-residue long with a 10-residue overlap) were tested for binding to CC40.8 mAb by ELISA. CC40.8 showed binding to the 95 th 15-mer peptide corresponding to the HCoV-HKU1 S2 stem-helix region (residue position range: 1231-1245). An unrelated antibody to dengue virus, DEN3, was a control. D . BLI binding of CC40.8 to the HCoV-HKU1 95 th 15-mer stem peptide (blue) and HCoV-HKU1 stem peptide variants with 5 additional residues either at the N-(20-mer: brick red) or C-(20-mer: orange) terminus or added at both termini (25-mer: red). CC40.8 showed strongest binding to the 25-residue stem peptide corresponding to HCoV-HKU1 S2 residues 1126-1150. BLI binding of CC40.8 to 25-mer stem peptides derived from all HCoV spikes. CC40.8 showed binding to the β-but not to the α-HCoV S2 stem peptides. E . SARS-CoV-2 S protein cartoon depicting the S2-stem epitope region in green at the base of the prefusion spike. F . Sequence conservation of CC40.8 stem-helix epitope on SARS-CoV-1/2, HCoV-HKU1 and HCoV-OC43 human β-CoV spikes. Conserved residues are highlighted with blue boxes, while similar residues in cyan boxes. An N-linked glycosylation site is indicated with a “#” symbol.

Article Snippet: Amino acid point mutations in SARS-CoV-2 spike encoding plasmids were made by using site-directed mutagenesis kit (NEB cat.# E0554S) according to the manufacturer’s instructions.

Techniques: Neutralization, Isolation, Binding Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Derivative Assay, Sequencing

Journal: bioRxiv

Article Title: A protective broadly cross-reactive human antibody defines a conserved site of vulnerability on beta-coronavirus spikes

doi: 10.1101/2021.03.30.437769

Figure Lengend Snippet: HeLa-SARS-CoV-2 & 1 Spike cells expressing nucleus-restricted RFP (red) are pre-incubated with no antibody, negative control antibody (DEN3) or CC40.8 for 1h, and mixed with HeLa-ACE2 cells expressing cytosolic GFP (green). Green syncytia were observed without antibody or with negative control (row 1, 2, 4 & 5), indicating widespread cell-cell fusion mediated by SARS-CoV-1 & 2 spike and hACE2; this was rescued by addition of CC40.8 (row 3 & 6). Hoechst was used to stain cell nuclei.

Article Snippet: Amino acid point mutations in SARS-CoV-2 spike encoding plasmids were made by using site-directed mutagenesis kit (NEB cat.# E0554S) according to the manufacturer’s instructions.

Techniques: Expressing, Incubation, Negative Control, Staining

Journal: bioRxiv

Article Title: A protective broadly cross-reactive human antibody defines a conserved site of vulnerability on beta-coronavirus spikes

doi: 10.1101/2021.03.30.437769

Figure Lengend Snippet: A . An overall view of the CC40.8 antibody and S2 stem-peptide interaction. Heavy and light chains of CC40.8 are shown in orange and yellow, respectively, whereas the SARS-CoV-2 stem peptide is in green. Hydrogen bonds and salt bridges are represented by black dashed lines. B . Details of interactions between CC40.8 and the SARS-CoV-2 stem peptide. Residues conserved in SARS-CoV-1, SARS-CoV-2, HCoV-HKU-1, and HCoV-OC43 are labeled with asterisks (*). C . Sequence conservation of CC40.8 stem-helix epitope on SARS-CoV-1/2, sarbecoviruses infecting other animal species, human β-CoVs and mouse hepatitis virus (MHV). The stem region forming the helix and the residues involved in interaction with CC40.8 antibody are indicated by red dots (cutoff distance = 4 Å), within which bigger dots indicate residues that are essential for CC40.8 interaction, where alanine scanning mutagenesis of such residue decreased neutralization IC50 for at least 10 folds or a complete knock-out (details are shown in ). Conserved residues are highlighted with blue boxes, while similar residues in cyan boxes. An N-linked glycosylation site is indicated with a “#” symbol. The region presents a helical secondary structure in the CC40.8/peptide structure is indicated on top of the panel. D . BLI binding of CC40.8 bnAb to SARS-CoV-2 stem-helix peptide and soluble spike alanine mutants spanning the whole epitope. The stem peptide or spike mutants that substantially affect CC40.8 bnAb binding are shown in colors. E . CC40.8 neutralization of SARS-CoV-2 and the stem-helix alanine mutants spanning the whole epitope. The virus mutants that substantially affect CC40.8 bnAb neutralization are shown in colors.

Article Snippet: Amino acid point mutations in SARS-CoV-2 spike encoding plasmids were made by using site-directed mutagenesis kit (NEB cat.# E0554S) according to the manufacturer’s instructions.

Techniques: Labeling, Sequencing, Mutagenesis, Neutralization, Knock-Out, Binding Assay

Journal: bioRxiv

Article Title: A protective broadly cross-reactive human antibody defines a conserved site of vulnerability on beta-coronavirus spikes

doi: 10.1101/2021.03.30.437769

Figure Lengend Snippet: A . Overall view of the CC40.8-peptide complex structure. Heavy and light chains of CC40.8 are shown in orange and yellow semi-transparent surfaces, respectively, where paratope regions are shown as cartoon. The SARS-CoV-2 stem-helix peptide is shown in green. B . Surface area of the SARS-CoV-2 stem peptide. Solvent exposed and buried areas were calculated with Proteins, Interfaces, Structures and Assemblies (PISA) . C . The SARS-CoV-2 stem peptide inserts into a hydrophobic groove formed by the heavy and light chains of CC40.8. Surfaces of CC40.8 are color-coded by hydrophobicity [calculated by Color h ( https://pymolwiki.org/index.php/Color_h )]. D . Electrostatic surface potential of the CC40.8 paratope. Electrostatic potential is calculated by APBS and PDB2PQR ( , ).

Article Snippet: Amino acid point mutations in SARS-CoV-2 spike encoding plasmids were made by using site-directed mutagenesis kit (NEB cat.# E0554S) according to the manufacturer’s instructions.

Techniques:

Journal: bioRxiv

Article Title: A protective broadly cross-reactive human antibody defines a conserved site of vulnerability on beta-coronavirus spikes

doi: 10.1101/2021.03.30.437769

Figure Lengend Snippet: A . Alignment of CC40.8 with the germline VH3-23 and VL3-10 sequences. Paratope residues [defined as BSA > 0 Å 2 as calculated by PISA are highlighted with yellow boxes. Somatically mutated residues as calculated by IgBLAST are highlighted in red. B . Detailed interactions between CC40.8 Fab and the SARS-CoV-2 stem peptide. Heavy and light chains of CC40.8 are shown in orange and yellow, while the SARS-CoV-2 stem peptide is in pale green. Hydrogen bonds and salt bridges are represented by black dashed lines. Somatically mutated residues are shown in red. Conserved residues among coronaviruses are indicated by asterisks (*).

Article Snippet: Amino acid point mutations in SARS-CoV-2 spike encoding plasmids were made by using site-directed mutagenesis kit (NEB cat.# E0554S) according to the manufacturer’s instructions.

Techniques:

Journal: bioRxiv

Article Title: A protective broadly cross-reactive human antibody defines a conserved site of vulnerability on beta-coronavirus spikes

doi: 10.1101/2021.03.30.437769

Figure Lengend Snippet: The upper panel shows the IC50 neutralization of CC40.8 bnAb with WT SARS-CoV-2 and spike mutant pseudoviruses and the BLI binding responses with WT SARS-CoV-2 soluble S protein and alanine mutants. SARS-CoV-2 receptor binding domain (RBD) antibody S309 was control for the spike binding assays. The IC50 fold change (n-fold) was calculated by dividing the mutant value by the WT value. For IC50, n-fold <0.3 are indicated in red, n-fold >5 in green. The middle and lower panels show BLI binding responses of CC40.8 antibody to WT and alanine mutants of the SARS-CoV-1/2 and HCoV-HKU1 stem peptides, respectively. For binding response values where the % change in binding (from WT peptide) is <80%, are indicated in yellow. Antibody S309 recognizing the RBD of both SARS-CoV-1 and SARS-CoV-2 was used as control. N/A, not available.

Article Snippet: Amino acid point mutations in SARS-CoV-2 spike encoding plasmids were made by using site-directed mutagenesis kit (NEB cat.# E0554S) according to the manufacturer’s instructions.

Techniques: Neutralization, Mutagenesis, Binding Assay

Journal: bioRxiv

Article Title: A protective broadly cross-reactive human antibody defines a conserved site of vulnerability on beta-coronavirus spikes

doi: 10.1101/2021.03.30.437769

Figure Lengend Snippet: A . A SARS-CoV-2 spike structure in pre-fusion state. The three protomers are shown in gray, pale green, and white, respectively with N-linked glycans represented by sticks. The 3-helix bundle stem region is highlighted in a blue-outlined box. Representative epitope residues of CC40.8 are shown in sticks. CC40.8 bnAb epitope is rich in hydrophobic residue. A cryo-EM structure of SARS-CoV-2 spike structure in pre-fusion state that contains the coordinates of the 3-helix bundle stem region (PDB: 6XR8, ) is shown here. B . SARS-CoV-2 spike pre-fusion structure superimposed with the structure of CC40.8 (orange/yellow) in complex with a SARS-CoV-2 S2 peptide. CC40.8 would clash with the other protomers of the spike protein in pre-fusion state. C . A putative neutralization mechanism of CC40.8. The S2 3-helix bundle region is shown in green, whereas heavy and light chains of CC40.8 in orange and yellow, respectively. Mechanism of neutralization model inspired by interaction of a mouse S2 stem antibody, B6, isolated from an S protein vaccinated animal that targets a similar stem epitope .

Article Snippet: Amino acid point mutations in SARS-CoV-2 spike encoding plasmids were made by using site-directed mutagenesis kit (NEB cat.# E0554S) according to the manufacturer’s instructions.

Techniques: Cryo-EM Sample Prep, Neutralization, Isolation

Journal: bioRxiv

Article Title: A protective broadly cross-reactive human antibody defines a conserved site of vulnerability on beta-coronavirus spikes

doi: 10.1101/2021.03.30.437769

Figure Lengend Snippet: A . CC40.8 was intraperitonially (i.p.) administered at 2 mg per animal dose into Syrian hamsters (average: 16.5 mg/kg). Control animals received 2 mg of control Zikv mAb1. Each group of five animals was challenged intranasally (i.n.) 12 hours after antibody infusion with 1 × 10 6 PFU of SARS-CoV-2. Animal weight was monitored daily as an indicator of disease progression and lung tissue was collected on day 5 for viral burden assessment. B . Percent weight change in CC40.8 or control antibody-treated animals after SARS-CoV-2 challenge. Percent weight change was calculated from day 0 for all animals. C . SARS-CoV-2 titers (PFU) as determined by plaque assay from lung tissue at day 5 after infection.

Article Snippet: Amino acid point mutations in SARS-CoV-2 spike encoding plasmids were made by using site-directed mutagenesis kit (NEB cat.# E0554S) according to the manufacturer’s instructions.

Techniques: Plaque Assay, Infection

Journal: bioRxiv

Article Title: A protective broadly cross-reactive human antibody defines a conserved site of vulnerability on beta-coronavirus spikes

doi: 10.1101/2021.03.30.437769

Figure Lengend Snippet: A . Heatmap showing ELISA binding reactivity profiles of convalescent COVID sera with 25-mer peptides corresponding to the CC40.8 bnAb S2 epitopes on human β-(SARS-CoV-2, SARS-CoV-1, MERS-CoV, HCoV-HKU1, HCoV-OC43) and α-(HCoV-NL63 and HCoV-229E) coronaviruses. The extent of binding is color coded with red indicating strong reactivity. CC40.8 mAb was the positive control for the binding assay and PBS-BSA solution served as the negative control. Six out of 60 COVID donors showed cross-reactive binding to various HCoV S peptides. B . ELISA-based alanine scan epitope mapping of convalescent COVID-19 sera from CC6, CC21, CC40, CC48, CC57 and CC57 donors with SARS-CoV-2 stem peptides (25mer). CC40 sera showed dependence on similar SARS-CoV-2 stem helix residues as the CC40.8 mAb. SARS-CoV-2 stem helix residue positions targeted (decrease in ELISA binding compared to WT stem peptide) by multiple cross-reactive COVID-19 sera are shown in grey. Five residues, F 1148 , E 1151 , L 1152 , Y 1155 and F 1156 were commonly targeted by the cross-reactive COVID-19 serum Abs. These residues form the stem-helix bnAb core epitope. C . SARS-CoV-2 neutralization by CC40.8 in presence of competing SARS-CoV-2 stem peptide. Neutralization of SARS-CoV-2 by CC40.8 mAb, CC40.8 mAb pre-incubated with SARS-CoV-2 stem peptide (60ug/ml) and stem peptide-only control. The SARS-CoV-2 stem peptide inhibits the neutralizing activity of CC40.8 mAb. D . SARS-CoV-2 neutralization by cross-reactive COVID-19 sera in presence of competing SARS-CoV-2 stem peptide. Neutralization of SARS-CoV-2 by sera from COVID-19 donors, CC6, CC21, CC40, CC48, CC57, CC57, sera pre-incubated with SARS-CoV-2 stem peptide (60ug/ml) and stem peptide only controls. The SARS-CoV-2 stem peptide had minimal effects on neutralization by stem-targeting COVID serum antibodies.

Article Snippet: Amino acid point mutations in SARS-CoV-2 spike encoding plasmids were made by using site-directed mutagenesis kit (NEB cat.# E0554S) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Positive Control, Negative Control, Neutralization, Incubation, Activity Assay

Journal: medRxiv

Article Title: Bivalent COVID-19 mRNA booster vaccination (BA.1 or BA.4/BA.5) increases neutralization of matched Omicron variants

doi: 10.1101/2023.04.20.23288813

Figure Lengend Snippet: (a) Antigenic maps of SARS-CoV-2 variants based on post-vaccination (V1/V2/V3-monovalent, n=31; V1/V2/V3/V4-monovalent, n=26; V1/V2/V3/V4-bivalent-BA.1, n=12; V1/V2/V3/V4-bivalent-BA.5, n=22) and Omicron breakthrough-infection sera, including subvariants BA.1, BA.2 and BA.4/BA.5 (n=22). Squares represent individual sera, circles SARS-CoV-2 variants. The x- and y-axes of the maps are antigenic distances, and each square represents a two-fold change in neutralization titer. (b) Cumulative antigenic distance scores. Boxes range from 25th to 75th percentile, whiskers show min and max, and horizontal lines the median. BTI, breakthrough infection. Scores were compared with Kruskal-Wallis tests with Dunn’s multiple comparison correction. ****p < 0.0001, ***p < 0.001.

Article Snippet: Nasopharyngeal swabs were analyzed with the mutation assays VirSNiP SARS-CoV-2 Spike S371L S373P, VirSNiP SARS-CoV-2 Spike 484A 486V and VirSNiP SARS-CoV-2 Spike L452R (TIB MOLBIOL, Berlin, Germany).

Techniques: Infection, Neutralization

Journal: Cell Reports Medicine

Article Title: Ongoing evolution of SARS-CoV-2 drives escape from mRNA vaccine-induced humoral immunity

doi: 10.1016/j.xcrm.2024.101850

Figure Lengend Snippet:

Article Snippet: pTwist-SARS-CoV-2 Spike-ΔC18 VOC-501Y.V2 (B.1.351) - All Mutations , This study , Addgene Plasmid ID 212381.

Techniques: Virus, Recombinant, Saline, Modification, Plasmid Preparation, Software, Control

Journal: Nature Communications

Article Title: Vaccination impairs de novo immune response to omicron breakthrough infection, a precondition for the original antigenic sin

doi: 10.1038/s41467-024-47451-w

Figure Lengend Snippet: Omicron SARS-CoV-2 is abbreviated as O and wild-type as WT throughout the figure. a Representative flow cytometry pseudocolor plots demonstrating the detection of T cells specific for the omicron spike (S) protein. Percentages of the parent populations are indicated within the gates. b Frequency of omicron-spike-specific CD4 T cells as a percentage of all T cells. The following numbers of biologically independent samples were included in each group: n(O-Inf) = 7, n(Vacc+O-Inf) = 36, n(Vacc) = 41. Frequency of omicron-not-wild-type-spike-specific CD4 T cells as ( c ) percentage of all T cells, ( e ) percentage of all omicron-spike-specific CD4 T cells. The following numbers of biologically independent samples were included in each group: n(O-Inf) = 7, n(Vacc+O-Inf) = 26, n(Vacc) = 36. d and h Time between the last exposure to SARS-CoV-2 antigens, either as vaccination or infection, and sampling time point for individuals included in different panels. The data is displayed as scatter plots with lines indicating mean and 95% confidence intervals. The numbers of biologically independent samples correspond to those used in each panel. f Omicron-spike-specific CD8 T cells as percentages of all T cells. The following numbers of biologically independent samples were included in each group: n(O-Inf) = 7, n(Vacc+O-Inf) = 35, n(Vacc) = 41. Frequency of omicron-not-wild-type-spike-specific CD8 T cells as ( g ) percentage of all T cells, ( i) percentage of all omicron-spike-specific CD8 T cells. The following numbers of biologically independent samples were included in each group: n(O-Inf) = 6, n(Vacc+O-Inf) = 12, n(Vacc) = 19. In panels b , c , e , f , g , and i the data is displayed as bar plots indicating the median and 95% confidence interval with individual data points. Differences between the groups were assessed using the Kruskal-Wallis test with Dunn’s correction for multiple testing. Source data including exact p values are provided as a Source Data file.

Article Snippet: B-cell-depleted PBMC fraction was seeded in 96-well U bottom plates and stimulated with two different pools of overlapping peptides: the first covering the mutated regions of the omicron SARS-CoV-2 spike protein (PepTivator® SARS-CoV-2 Prot_S B.1.1.529/BA.1 Mutation Pool, Miltenyi Biotec, 130-129-928) and the second covering conserved regions of the spike (PepTivator® SARS-CoV-2 Prot_S B.1.1.529/BA.1 WT Reference Pool, Miltenyi Biotec, 130-129-927).

Techniques: Flow Cytometry, Infection, Sampling

Journal: Journal of Biochemistry

Article Title: SARS-CoV-2 spike protein binding selectively accelerates substrate-specific catalytic activity of ACE2

doi: 10.1093/jb/mvab041

Figure Lengend Snippet: Binding of the SARS-CoV-2 spike protein increases the rate of ACE2 activity. ( A ) Kinetic curves showing the effect of full-length SARS-CoV-2 spike on ACE2 activity. ( B ) Kinetic curves showing the effect of SARS-CoV-1 and SARS-CoV-2 RBD spike on ACE2 activity. ( C ) Michaelis–Menten plot showing effect of SARS-CoV-1 and SARS-CoV-2 RBD on catalytic activity of ACE2. k obs , observed rate constant. ( D ) Catalytic rate ( k cat ) and K M of ACE2 in the absence or presence of SARS-CoV-1 and SARS-CoV-2 RBD spike protein [mean (95% confidence intervals)]. Pseudosubstrate concentration in (A) and (B) is 20 µM. Assays were conducted in two biological replicates.

Article Snippet: Recombinant SARS-CoV-2 RBD spike mutant (Y505C) was obtained from Sino Biological (cat# 40592-V08H72, Sino Biological US Inc., PA, USA).

Techniques: Binding Assay, Activity Assay, Concentration Assay

Journal: Journal of Biochemistry

Article Title: SARS-CoV-2 spike protein binding selectively accelerates substrate-specific catalytic activity of ACE2

doi: 10.1093/jb/mvab041

Figure Lengend Snippet: SARS-CoV-2 spike protein accelerates the activity of ACE2 in a substrate-dependent manner. Kinetic curves showing the effect of SARS-CoV-2 spike RBD binding on ACE2 activity in the presence of ( A ) pseudosubstrate MCA-YVADAPK(Dnp); ( B ) angiotensin II mimic [MCA-DRVYIHPK(Dnp)]; ( C ) apelin 13 mimic [MCA-QRPRLSHKGPMPK(Dnp)]; ( D ) des-Arg9-bradykinin mimic [MCA-RPPGFSPK(Dnp)] ( E ) angiotensin I mimic [MCA-DRVYIHPFK(Dnp)]; ( F ) dynorphin A mimic [MCA-YGGFLRRIRPKLK(Dnp)] substrates; ( G ) Michaelis–Menten plot showing effect of SARS-CoV-2 RBD on catalytic activity of ACE2 in the presence of des-Arg9-bradykinin mimic substrate. ( H ) Catalytic rate ( k cat ) and K M of ACE2 in the absence or presence of SARS-CoV-2 RBD spike protein and des-Arg9-bradykinin mimic substrate [mean (95% confidence intervals)]. Substrate concentration in (A)–(F) is 20 µM. Assays were conducted in two biological replicates.

Article Snippet: Recombinant SARS-CoV-2 RBD spike mutant (Y505C) was obtained from Sino Biological (cat# 40592-V08H72, Sino Biological US Inc., PA, USA).

Techniques: Activity Assay, Binding Assay, Concentration Assay

Journal: Journal of Biochemistry

Article Title: SARS-CoV-2 spike protein binding selectively accelerates substrate-specific catalytic activity of ACE2

doi: 10.1093/jb/mvab041

Figure Lengend Snippet: Binding of the heat-inactivated SARS-CoV-2 viral particles accelerates ACE2 catalytic activity. ( A – C ) Kinetic curves showing the effect of different concentrations of heat-inactivated SARS-CoV-2 on ACE2 activity in the presence of des-Arg9-bradykinin mimic [MCA-RPPGFSPK(Dnp)] substrate (A); MCA-YVADAPK(Dnp) substrate (B); and angiotensin II mimic [MCA-DRVYIHPK(Dnp)] substrate (C). Substrate concentration in (A)–(C) is 20 µM. Assays were conducted in two biological replicates.

Article Snippet: Recombinant SARS-CoV-2 RBD spike mutant (Y505C) was obtained from Sino Biological (cat# 40592-V08H72, Sino Biological US Inc., PA, USA).

Techniques: Binding Assay, Activity Assay, Concentration Assay

Journal: Genes & Diseases

Article Title: Intranasal boosting with RBD-HR protein vaccine elicits robust mucosal and systemic immune responses

doi: 10.1016/j.gendis.2023.06.035

Figure Lengend Snippet: Immune sera inhibited the binding of SARS-CoV-2 pseudoviruses to hACE2. (A–C) Fourteen days after the last immunization, the mouse sera were diluted to 1:90. (A) Representative graphs of fluorescence microscopy illustrating the neutralization of immune sera to WT SARS-CoV-2 pseudovirus. Scale bar = 200 μm. (B) Representative graphs of flow cytometry depicting the neutralization of immune sera to WT SARS-CoV-2 pseudovirus. (C) Neutralizing activities of immune sera to WT SARS-CoV-2 pseudovirus characterized by the relative intensity of eGFP. (D–E) pVNT50 of immune sera against SARS-CoV-2 pseudoviruses, including WT, B.1.617, BA.1, BA.2, BA.3, and BA.4/5. Results were expressed as mean ± SEM, and P values were calculated by one-way ANOVA. n = 6, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001; ns: P > 0.05.

Article Snippet: The inhibition of specific antibodies on RBD-Fc binding to hACE2-expressing cells was observed by flow cytometry as previously reported., Prototype and mutant recombinant SARS-CoV-2 RBD-Fc, including WT, B.1.617.2, and BA.1 (Sino Biological, China; Acro, China), were preincubated with 1:90 diluted sera at 37 °C for 0.5 h in 96-well plates.

Techniques: Binding Assay, Fluorescence, Microscopy, Neutralization, Flow Cytometry

Journal: iScience

Article Title: SARS-CoV-2 antigen-carrying extracellular vesicles activate T cell responses in a human immunogenicity model

doi: 10.1016/j.isci.2023.108708

Figure Lengend Snippet:

Article Snippet: Treatments with SARS-CoV-2 peptides (PepTivator SARS-CoV-2 Prot_S; Miltenyi Biotech; 130-126-700) and recombinant SARS-CoV-2 Spike protein (with K986P, V987P, R682S and R685S mutations as well as C-term His tag; R&D Systems, 10549-CV) were included as positive controls.

Techniques: Recombinant, Transduction, Transfection, Expressing, Electron Microscopy, Membrane, Bicinchoninic Acid Protein Assay, Enzyme-linked Immunosorbent Assay, Binding Assay, Plasmid Preparation, Software, Sequencing