mammalian rbd  (GenScript corporation)

Journal: Journal of Translational Medicine

Article Title: Potential inhibitors for blocking the interaction of the coronavirus SARS-CoV-2 spike protein and its host cell receptor ACE2

doi: 10.1186/s12967-022-03501-9

Figure Lengend Snippet: TS-984 can inhibit the SARS-COV-2 pseudo virus entering the Capan2 with ACE2 overexpression. A TS-984 can greatly reduce the EGFP/mCherry signal ratio. [*P < 0.05 and **P < 0.01 in comparison to control group]. B The 10× fluorescence image show that TS984 can inhibit the entering of pseudoviurs (green) into the Capan2 with ACE2 overexpression. (Scar bar, 200 μm)

Article Snippet: In brief, ACE2 and SARS-Cov-2 S-protein were prepared at multiple concentrations with a PBS containing 0.1% BSA, 5 μl ACE2 (Mammalian, C-Fc, DRA36, Novoprotein) and 5 μl SARS-Cov-2 S-protein RBD (Mammalian, C-6His, C05Y, Novoprotein).

Techniques: Over Expression, Fluorescence

Journal: Journal of Translational Medicine

Article Title: Potential inhibitors for blocking the interaction of the coronavirus SARS-CoV-2 spike protein and its host cell receptor ACE2

doi: 10.1186/s12967-022-03501-9

Figure Lengend Snippet: TS984 can inhibit the SARS-COV-2 pseudo virus entering the 293T with ACE2 overexpression. A TS984 can greatly reduce the EGFP/mCherry signal ratio. [*P < 0.05 and **P < 0.01 in comparison to control group]. B The 10× fluorescence image show that TS984 can inhibit the entering of pseudoviurs (green) into the 293T with ACE2 overexpression. (Scar bar, 200 μm)

Article Snippet: In brief, ACE2 and SARS-Cov-2 S-protein were prepared at multiple concentrations with a PBS containing 0.1% BSA, 5 μl ACE2 (Mammalian, C-Fc, DRA36, Novoprotein) and 5 μl SARS-Cov-2 S-protein RBD (Mammalian, C-6His, C05Y, Novoprotein).

Techniques: Over Expression, Fluorescence

Journal: Cell

Article Title: In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies

doi: 10.1016/j.cell.2021.06.021

Figure Lengend Snippet: SARS-CoV-2 receptor-binding domain (RBD) and N-terminal domain (NTD) Abs mediate enhancement of infection (A and B) Timeline of blood sampling, plasmablasts and/or antigen-specific memory B cells (MBC) sorting, and Ab isolation from convalescent (A) SARS-CoV-2 and (B) SARS-CoV donors. (C) Summary of number and specificity of Abs isolated from each donor. (D and E) In vitro neutralization curves for NTD infection-enhancing Abs against (D) pseudotyped SARS-CoV-2 D614G in 293T-hACE2 cells, and (E) replication-competent nano-luciferase (nLuc) SARS-CoV-2 in Vero cells. (F–J) FcγR-dependent pseudotyped SARS-CoV-2 infection enhancement when RBD Abs or mock medium control was added to (F) parental TZM-bl cells, and TZM-bl cells stably expressing human FcγR receptors (G) FcγRI, (H) FcγRIIa, (I) FcγRIIb or (J) FcγRIII. (K and L) The effect of RBD Ab fragment antigen-binding regions (Fabs) on pseudotyped SARS-CoV-2 D614G infection was tested in (K) FcγRI-expressing TZM-bl cells and (L) FcγRIIb-expressing TZM-bl cells. Data are represented as mean ± SEM. Three or four independent experiments were performed, and representative data are shown. See also Figure S1 .

Article Snippet: SARS-CoV-2 Spike RBD from mammalian cell 293 , Sino Biological , Cat #40592-V08H.

Techniques: Binding Assay, Infection, Sampling, Isolation, In Vitro, Neutralization, Luciferase, Stable Transfection, Expressing

Journal: Cell

Article Title: In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies

doi: 10.1016/j.cell.2021.06.021

Figure Lengend Snippet: Isolation of SARS-CoV-2-reactive Abs from single cell-sorted plasmablasts and memory B cells of SARS-CoV-2 and SARS-CoV-1 convalescent donors, related to (A) Symptom severity scores of the COVID-19 convalescent donor. The method to determine severity score is in supplementary online material. Red arrows indicate the blood sampling time points that we used to isolate Abs. (B) Viral load from nasopharyngeal (NP) swabs. (C) Serum micro-neutralization titer. Micro-Neutralization titers were defined as the highest serum dilution that neutralize all the virus, or 99% inhibitory concentration (IC 99 ). (D) Flow cytometry gating strategy for unbiased plasmablasts sorting or antigen specific-memory B cells sorting. At day 11 and day 15 post onset of COVID-19 symptom, plasmablasts (CD14 - /CD16 - /CD3 - /CD235a - /CD19 + /CD20 low /IgD - /CD27 high /CD38 high ) from a SARS-CoV-2 donor. Antigen specific B cells from SARS-CoV-1 and SARS-CoV-2 donors were sorted with different combinations of the SARS-CoV-2 S-2P, RBD, NTD probes. Representative data for sorting Spike double positive, Spike + or NTD + , as well as RBD + or NTD + subsets were shown. (E-H) RBD Ab neutralization activity. (E) Proportion of SARS-CoV-2 RBD Abs (n = 81) that exhibited detectable neutralization in the microneutralization assay. (F) Neutralization IC 50 and IC 80 of RBD neutralizing Abs (NAbs) against pseudotyped SARS-CoV-2. (G) Microneutralization titer, plaque reduction neutralization test (PRNT) IC 50 and IC 80 of RBD NAbs against replication-competent SARS-CoV-2. Microneutralization titer was defined as the lowest Ab concentration that neutralized all the virus, or 99% inhibitory concentration (IC 99 ). Abs with undetectable microneutralization titers are shown as gray symbols and nAbs are represented by blue symbols. (H) RBD NAbs blocking of ACE2 binding to SARS-CoV-2 Spike (S) protein. Blocking titer is shown as IC 50 . (I-J) Correlation analysis of RBD Abs between neutralization and ACE2 blocking activities. Spearman correlation analysis were performed for (I) ACE2 blocking IC 50 versus PV neutralization IC 50 , as well as (J) for ACE2 blocking IC 50 versus SARS-CoV-2 neutralization titers (indicated by the lowest concentration that shows no CPE). Purified RBD Abs in and that have pseudovirus neutralization data (n = 59) or SARS-CoV-2 micro-neutralization assay data (n = 80) were used in this analysis. P values (p)and correlation coefficients (r) are indicated for each figure. (K-M) Neutralization activity of NTD Abs. (K) Proportion of SARS-CoV-2 NTD Abs (n = 41) that exhibited detectable neutralization in the microneutralization assay. (L) Neutralization IC 50 and IC 80 of NTD neutralizing Abs against pseudotyped SARS-CoV-2. (M) Microneutralization titer, PRNT IC 50 and IC 80 of NTD neutralizing Abs against replication-competent SARS-CoV-2. Abs with undetectable microneutralization titers are shown as gray symbols and neutralizing Abs are represented by orange symbols. Horizontal bars represent the geometric means for each group of Abs.

Article Snippet: SARS-CoV-2 Spike RBD from mammalian cell 293 , Sino Biological , Cat #40592-V08H.

Techniques: Isolation, Sampling, Neutralization, Concentration Assay, Flow Cytometry, Activity Assay, Microneutralization Assay, Plaque Reduction Neutralization Test, Blocking Assay, Binding Assay, Purification

Journal: Cell

Article Title: In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies

doi: 10.1016/j.cell.2021.06.021

Figure Lengend Snippet: Structural and phenotypic characterization of infection-enhancing and non-infection-enhancing RBD and NTD Abs. (A) Summary of Ab epitope, binding, and neutralizing or infection-enhancing activity in ACE2-positive/FcγR-negative cells or ACE2-negative/FcγR-positive cells. Ab functions are color-coded based on the key shown at the right. MN titer, micro-neutralization titer; ND, not determined. (B–E) 3D reconstruction of negative stain electron microscopy images of stabilized SARS-CoV-2 S ectodomain trimers (S-2P; gray) bound to the Fabs (various colors) of (B and D) infection-enhancing or (C and E) non-infection-enhancing RBD or NTD antibodies. See also and .

Article Snippet: SARS-CoV-2 Spike RBD from mammalian cell 293 , Sino Biological , Cat #40592-V08H.

Techniques: Infection, Binding Assay, Activity Assay, Neutralization, Staining, Electron Microscopy

Journal: Cell

Article Title: In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies

doi: 10.1016/j.cell.2021.06.021

Figure Lengend Snippet: Binding and neutralization activities of down-selected SARS-CoV-2 Abs, related to Figure 2 (A-D) ELISA binding curves of down-selected Abs. Different SARS-CoV-2 or other CoV viral antigens were coated on plates and detected with serial diluted (A) RBD infection-enhancing Abs, (B) RBD non-infection-enhancing Abs, (C) NTD infection-enhancing Abs, and (D) NTD non-infection-enhancing Abs. (E-F) Neutralization curves for RBD Abs against pseudotyped (E) and replication-competent (F) SARS-CoV-2. (G-H) Neutralization curves for NTD Abs against pseudotyped (G) and replication-competent (H) SARS-CoV-2. (I-L) Neutralization curves for cross-neutralizing Abs against pseudotyped (I) and replication-competent (J) SARS-CoV-2, SARS-CoV-1 nanoluciferase (nLuc) virus (L), and Bat WIV1-CoV nLuc virus (L).

Article Snippet: SARS-CoV-2 Spike RBD from mammalian cell 293 , Sino Biological , Cat #40592-V08H.

Techniques: Binding Assay, Neutralization, Enzyme-linked Immunosorbent Assay, Infection

Journal: Cell

Article Title: In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies

doi: 10.1016/j.cell.2021.06.021

Figure Lengend Snippet: Simultaneous binding of infection-enhancing and non-infection-enhancing Abs to individual S trimers (A) Cross-blocking activity of RBD and NTD-neutralizing Abs tested by surface plasmon resonance (SPR). S-2P was captured by one Ab (y axis) followed by binding by the second Ab (x axis). (B) 3D reconstruction of simultaneous recognition of SARS-CoV-2 S-2P by two RBD Abs DH1041+DH1047 or DH1043+DH1047. (C) Cross-blocking activity of neutralizing or infection-enhancing NTD Abs tested by SPR and shown as in (A). (D–F) 3D reconstruction of SARS-CoV-2 S-2P simultaneously bound (D) NTD Abs DH1053 and DH1050.1, (E) RBD infection-enhancing Ab and a NTD non-infection-enhancing Ab, or (F) triple-Ab combinations of RBD Ab DH1043, RBD Ab DH1047, and either NTD Ab DH1051 (left) or DH1050.1 (right). (G and H) RBD Ab neutralization of SARS-CoV-2 D614G pseudovirus infection of 293T/ACE2 cells in the presence of 132 or 1,325 fold excess of infection-enhancing NTD Ab DH1052. See also .

Article Snippet: SARS-CoV-2 Spike RBD from mammalian cell 293 , Sino Biological , Cat #40592-V08H.

Techniques: Binding Assay, Infection, Blocking Assay, Activity Assay, SPR Assay, Neutralization

Journal: Cell

Article Title: In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies

doi: 10.1016/j.cell.2021.06.021

Figure Lengend Snippet: In vitro analysis of human Abs and SARS-CoV-2-infected serum samples, related to , and (A-C) Effect of combining infection-enhancing RBD and NTD Abs on SARS-CoV-2 pseudovirus infection in ACE2-expressing cells. The infection-enhancing NTD Ab DH1052 was tested alone (A) or mixed with infection-enhancing RBD Abs DH1041 (B) or DH1043 (C) in 1:13 ratio or 1:13250 ratio, respectively. The NTD:RBD Ab mixtures (orange), as well as RBD Ab alone (blue), were five-fold serially diluted and tested for neutralization against SARS-CoV-2 D614G pseudovirus in 293T/ACE2 cells. (D-F) Comparison of RBD and NTD directed serum Ab responses in SARS-CoV-2 infected humans. (D) Serum IgG binding titers to RBD (blue) and NTD (salmon) as measured by ELISA as log area-under-curve (AUC). Each symbol represents an individual study participant, with the mean binding titer for the visit day shown as a black horizontal bar. (E) Percent decrease in binding to NTD relative to RBD binding titer. Each symbol represents the change in binding titer for an individual study subject. Mean decrease is shown as a black horizontal bar. (F) Serum blocking of RBD neutralizing Ab DH1041 (blue) or NTD neutralizing Ab DH1050.1 (salmon), or non-neutralizing Ab DH1052 (burgundy) binding to SARS-CoV-2 spike. Black symbols show individual study participants. Mean blocking percentage for the visit day is shown as a filled bar. (G-H) Neutralization activities of neutralizing and enhancing Abs against wild-type (WT) and -mouse-adapted SARS-CoV-2. (G) NTD neutralizing Abs DH1050.1, RBD neutralizing and enhancing Abs DH1041 were tested for neutralization activities against WT virus, mouse-adapted 2AA MA virus, and mouse-adapted MA10 virus in live virus neutralization assay. CH65 Ab was used as a control. Mean value of neutralization (%) from duplicate wells were shown. (H) NTD enhancing Ab DH1052 and control Ab CH65 were tested for neutralization activities against WTvirus, mouse adapted 2AA MA virus, and mouse-adapted MA10 virus in live virus neutralization assay. Mean values of neutralization (%) from duplicate wells were shown.

Article Snippet: SARS-CoV-2 Spike RBD from mammalian cell 293 , Sino Biological , Cat #40592-V08H.

Techniques: In Vitro, Infection, Expressing, Neutralization, Binding Assay, Enzyme-linked Immunosorbent Assay, Blocking Assay

Journal: Cell

Article Title: In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies

doi: 10.1016/j.cell.2021.06.021

Figure Lengend Snippet: NTD Ab DH1052 does not always enhance SARS-CoV-2 replication or disease in vivo (A–F) DH1052 passive immunization and murine SARS-CoV-2 challenge (A) study design, (B) body weight, (C) survival, (D) hemorrhagic scores, (E) lung viral titers, and (F) SARS-CoV-2 envelope (E) and nucleocapsid (N) gene subgenomic RNA (sgRNA). (G–Q) Reduction of SARS-CoV-2 replication and disease in cynomolgus macaques by prophylactic administration of an NTD-neutralizing Ab DH1050.1 or an NTD in vitro infection-enhancing Ab DH1052. (G) DH1050.1 and DH1052 prophylaxis cynomolgus macaque (n = 5 per group) study design. CH65 was used as a negative control Ab. (H and I) Serum human IgG concentrations at (H) day −5 and (I) day 2. (J and K) Day 2 serum neutralization titers shown as the reciprocal serum dilution that inhibits 50% (ID 50 ) of (J) pseudotyped SARS-CoV-2 replication in 293T/ACE2 cells or (K) SARS-CoV-2 replication in Vero cells. (L and M) Lung histopathology 4 days post-infection. Lung sections were scored for (L) inflammation by hematoxylin and eosin (H&E) staining, and for (M) the presence of SARS-CoV-2 nucleocapsid by immunohistochemistry (IHC) staining. (N–Q) Viral load quantified as SARS-CoV-2 E gene sgRNA and N gene sgRNA in (N and O) bronchoalveolar lavage (BAL) or (P and Q) nasal swab fluid on day 2 and day 4 post challenge. LOD, limit of detection. Statistical significance in all the panels were determined using Wilcoxon rank-sum exact test. Horizontal bars are the group mean except in (J and K) where geometric mean is shown. Error bars indicate standard error of the mean. Asterisks show the statistical significance between the indicated group and CH65 control group: ns, not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. See also , , and .

Article Snippet: SARS-CoV-2 Spike RBD from mammalian cell 293 , Sino Biological , Cat #40592-V08H.

Techniques: In Vivo, In Vitro, Infection, Negative Control, Neutralization, Histopathology, Staining, Immunohistochemistry

Journal: Cell

Article Title: In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies

doi: 10.1016/j.cell.2021.06.021

Figure Lengend Snippet: RBD Abs that mediate FcγR-dependent infection enhancement in vitro protect non-human primates from SARS-CoV-2 challenge (A) Cynomolgus macaques (n = 5 per group) RBD Ab SARS-CoV-2 challenge study design. DH1041, DH1043, DH1046, DH1047, or an irrelevant CH65 were infused into macaques. (B and C) Serum human IgG concentrations at day −5 (B) and day 2 (C). (D and E) Day 2 serum neutralization titers shown as the reciprocal serum dilution that inhibits 50% (ID 50 ) of (D) pseudotyped SARS-CoV-2 replication in 293T/ACE2 cells or (E) SARS-CoV-2 replication in Vero cells. (F and G) Lung histopathology for (F) inflammation by H&E staining and (G) the presence of SARS-CoV-2 nucleocapsid by IHC staining 4 days post-challenge. (H–K) Viral load quantified as SARS-CoV-2 E gene sgRNA and N gene sgRNA in (H and I) bronchoalveolar lavage (BAL) or (J and K) nasal swab fluid on day 2 and day 4 post-challenge. Statistical significance in all the panels were determined using Wilcoxon rank-sum exact test. Horizontal bars are the group mean except in (D and E) where group geometric mean is shown. Asterisks show the statistical significance between indicated group and CH65 control group: ns, not significant, ∗ p < 0.05, ∗∗ p < 0.01. See also , , and .

Article Snippet: SARS-CoV-2 Spike RBD from mammalian cell 293 , Sino Biological , Cat #40592-V08H.

Techniques: Infection, In Vitro, Neutralization, Histopathology, Staining, Immunohistochemistry

Journal: Cell

Article Title: In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies

doi: 10.1016/j.cell.2021.06.021

Figure Lengend Snippet: Cryo-electron microscopy of neutralizing and non-neutralizing Abs in complex with SARS-CoV-2 Spike ectodomain Structures of SARS-CoV-2 S protein in complex with RBD Abs (A) DH1041 (red), (B) DH1043 (pink), (C) DH1047 (magenta), (D) neutralizing NTD Ab DH1050.1 (blue), and (E) infection-enhancing NTD Ab DH1052 (green). Each Ab is bound to S-2P shown in gray with its RBM colored purple blue. (Right) Zoomed-in views of the Ab interactions with S-2P trimers. The Ab complementarity determining (CDR) loops are colored: HCDR1 yellow, HCDR2 limon, HCDR3 cyan, LCDR1 orange, LCDR2 wheat and LCDR3 light blue. See also .

Article Snippet: SARS-CoV-2 Spike RBD from mammalian cell 293 , Sino Biological , Cat #40592-V08H.

Techniques: Electron Microscopy, Infection

Journal: Cell

Article Title: In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies

doi: 10.1016/j.cell.2021.06.021

Figure Lengend Snippet: Lung histopathology of Ab-treated and SARS-CoV-2 challenged cynomolgus macaques, related to and (A) Representative images of hematoxylin and eosin (H&E) staining and SARS-CoV-2 antigen immunohistochemistry (IHC) staining from each group. All images were taken at 10x magnification. The images in this figure are representative of the average severity of pathologic processes observed and recorded during microscopic evaluation. Red arrows indicate SARS-CoV-2 infection foci. (B) Following microscopic evaluation of DH1052, 1 animal (BB536A) out of 5 animals in this group exhibited histologic features that was substantially more severe than the rest of the cohort and may suggest some degree of Ab-mediated disease enhancement. The features were characterized by prominent perivascular mononuclear inflammation ( ∗ ) and a substantial amount of perivascular and alveolar edema (fluid; X). These findings suggest a vaso-centric process with some degree of altered vascular permeability. The remaining 4 animals in DH1052 group had inflammatory changes that ranged from minimal to moderate severity and more infiltrates were mixed and predominantly polymorphonuclear with lesser mononuclear cell involvement and present in the alveolar spaces. (C-E) Expression of macrophage activation markers in macaque lung tissues. An animal from the CH65 control group (C), the DH1052-treated animal (BB536A) that exhibited substantially more severe lung inflammation (D), and an animal from the NTD NAb DH1050.1 group (E) were selected for Immunohistochemistry (IHC) staining. Immunohistochemical staining was performed using MHCII, CD68, IBA1 and CD163 to detect classically activated macrophages (M1) and/or alternatively activated macrophages (M2). CD11b is a macrophage/monocyte marker and CD3 is a T cell marker. All images are 10x magnification; scale bars = 100μm.

Article Snippet: SARS-CoV-2 Spike RBD from mammalian cell 293 , Sino Biological , Cat #40592-V08H.

Techniques: Histopathology, Staining, Immunohistochemistry, Infection, Permeability, Expressing, Activation Assay, Immunohistochemical staining, Marker

Journal: Cell

Article Title: In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies

doi: 10.1016/j.cell.2021.06.021

Figure Lengend Snippet: High-dose NTD enhancing Ab DH1052 does not enhance SARS-CoV-2 replication or disease in vivo , related to Figure 5 (A) Diagram of the macaque study design showing cynomolgus macaques (n = 5 per group) were infused with high dose (30 mg/kg body weight) DH1052 or an irrelevant control CH65 Ab 3 days before 10 5 PFU of SARS-CoV-2 challenge via intranasal and intratracheal routes. Viral load including viral RNA and subgenomic RNA (sgRNA) were measured at the indicated pre-challenge and post-challenge time points. Lungs were harvested on Day 4 post-challenge for histopathology analysis. (B-D) SARS-CoV-2 (B) E gene sgRNA, (C) N gene sgRNA and (D) E gene total viral RNA in bronchoalveolar lavage (BAL) on Day 2 and Day 4 post challenge. (E-G) SARS-CoV-2 (E) E gene sgRNA, (F) N gene sgRNA and (G) E gene total viral RNA in nasal swab on Day 2 and Day 4 post challenge. (H-I) Lung inflammation. Sections of the left caudal (Lc), right middle (Rm), and right caudal (Rc) lung were evaluated and scored for the presence of inflammation by hematoxylin and eosin (H&E) staining. (H) Summary of inflammation scores. Symbols indicate the sums of Lc, Rm, and Rc scores in each animal. (I) Representative images of lung H&E staining. (J-K) Immunohistochemistry (IHC) staining for the presence of SARS-CoV-2 nucleocapsid in lungs. (J) Summary of IHC scores. Symbols indicate the sums of Lc, Rm, and Rc scores in each animal. (K) Representative images of lung IHC staining. Red arrows indicate SARS-CoV-2 infection foci. LOD, limit of detection. Horizontal bars are the group mean except in (C) where group geometric mean is shown. Statistical significance in all the panels were determined using Wilcoxon rank sum exact test. Asterisks show the statistical significance between the indicated group and CH65 control group: ns, not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: SARS-CoV-2 Spike RBD from mammalian cell 293 , Sino Biological , Cat #40592-V08H.

Techniques: In Vivo, Histopathology, Staining, Immunohistochemistry, Infection

Journal: Cell

Article Title: In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies

doi: 10.1016/j.cell.2021.06.021

Figure Lengend Snippet: RBD Abs that mediate FcγR-dependent infection enhancement in vitro protect mice from SARS-CoV-2 or bat WIV1-CoV challenge (A and B) Protection of BALB/c mice (n = 5 per group) from mouse-adapted SARS-CoV-2 (SARS-CoV-2 2AA MA) by (A) prophylactic or (B) therapeutic RBD and/or NTD Ab administration. Ab CH65 served as a negative control. Titers of infectious virus in the lung were examined 48 h post-infection. (C) Maximum likelihood tree of Spike amino acid sequences for human, bat, and pangolin coronaviruses. (D) Monoclonal RBD, NTD and S2 Ab ELISA binding titer for soluble S protein ectodomains from human and animal coronaviruses. Titers are log area-under-the-curve (AUC). (E) SARS-CoV and bat WIV1-CoV cross-neutralization titers for cross-reactive RBD and S2 Abs. (F and G) Protection of HFH4-hACE2-transgenic mice (n = 5 per group) from SARS-related bat WIV1-CoV challenge by (A) prophylactic or (B) therapeutic RBD Ab administration. Lung viral titers were examined at 48 h post-infection. Statistical significance in all the panels were determined using Wilcoxon rank-sum exact test. Horizontal bars are the group mean. Asterisks show the statistical significance between the indicated group and CH65 control group: ns, not significant, ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: SARS-CoV-2 Spike RBD from mammalian cell 293 , Sino Biological , Cat #40592-V08H.

Techniques: Infection, In Vitro, Negative Control, Enzyme-linked Immunosorbent Assay, Binding Assay, Neutralization, Transgenic Assay

Journal: Cell

Article Title: In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies

doi: 10.1016/j.cell.2021.06.021

Figure Lengend Snippet: Different doses of a cross-neutralizing Ab DH1047 treatments do not enhance SARS-CoV-2 replication in vivo , related to Figure 7 (A) Diagram of the macaque study design. Cynomolgus macaques (n = 5 per group) were infused with DH1047 at the dose of 10 mg/kg, 5 mg/kg, 1 mg/kg, 0.1 mg/kg weight. Macaques treated with 10 mg/kg weight of DH65 Ab were set as the control group. Three days post-infusion, 10 5 PFU of SARS-CoV-2 challenge via intranasal and intratracheal routes. Viral load including viral RNA and subgenomic RNA (sgRNA) were measured at the indicated pre-challenge and post-challenge time points. Lungs were harvested on Day 4 post-challenge for histopathology analysis. (B) Serum human IgG concentrations at Day 2. (C) Day 2 serum neutralization titers shown as the reciprocal serum dilution that inhibits 50% (ID 50 ) of SARS-CoV-2 replication in Vero cells. (D-E) SARS-CoV-2 (D) E gene sgRNA and (E) N gene sgRNA in bronchoalveolar lavage (BAL) on Day 2 and Day 4 post challenge. (F-G) SARS-CoV-2 (F) E gene sgRNA and (G) N gene sgRNA in nasal swab on Day 2 and Day 4 post challenge. (H-I) Lung inflammation. Sections of the left caudal (Lc), right middle (Rm), and right caudal (Rc) lung were evaluated and scored for the presence of inflammation by hematoxylin and eosin (H&E) staining. (H) Summary of inflammation scores. Symbols indicate the sums of Lc, Rm, and Rc scores in each animal. (I) Representative images of lung H&E staining. (J-K) Immunohistochemistry (IHC) staining for the presence of SARS-CoV-2 nucleocapsid in lungs. (J) Summary of IHC scores. Symbols indicate the sums of Lc, Rm, and Rc scores in each animal. (K) Representative images of lung IHC staining. Red arrows indicate SARS-CoV-2 infection foci. LOD, limit of detection. Horizontal bars are the group mean except in (C) where group geometric mean is shown. Statistical significance in all the panels were determined using Wilcoxon rank sum exact test. Asterisks show the statistical significance between the indicated group and CH65 control group: ns, not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: SARS-CoV-2 Spike RBD from mammalian cell 293 , Sino Biological , Cat #40592-V08H.

Techniques: In Vivo, Histopathology, Neutralization, Staining, Immunohistochemistry, Infection

Journal: Cell

Article Title: In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies

doi: 10.1016/j.cell.2021.06.021

Figure Lengend Snippet:

Article Snippet: SARS-CoV-2 Spike RBD from mammalian cell 293 , Sino Biological , Cat #40592-V08H.

Techniques: Recombinant, Staining, Random Hexamer Labeling, Luciferase, Cell Culture, Lysis, Multiplex Assay, Reporter Gene Assay, Binding Assay, Expressing, Transgenic Assay, Sequencing, Software