Journal: Lipids in Health and Disease

Article Title: Klotho ameliorates oxidized low density lipoprotein (ox-LDL)-induced oxidative stress via regulating LOX-1 and PI3K/Akt/eNOS pathways

doi: 10.1186/s12944-017-0447-0

Figure Lengend Snippet: Effect of Klotho on cellular viability in HUVECs. Cells were exposed to various concentrations of ox-LDL (25, 50, 100, and 200 μg/ml) for 24 h. a HUVEC morphology was observed under an inverted phase contrast microscope (×10) following 24 h of ox-LDL treatment. Typical cellular fragmentations, vacuoles, and debris are arrowed. (Bar = 50 μm) ( b ) Cellular viability was detected by MTT assay. c and d SOD enzymatic activity and MDA levels in HUVECs were analyzed using commercially available assay kits. e HUVECs were pretreated with 100, 200, 400, and 800 pM of Klotho for 1 h, and then incubated with ox-LDL (50 μg/ml) for 24 h. Cellular viability was detected by MTT assay. Data are shown as mean ± S.D. ( n = 3). Statistical differences are expressed as ## p < 0.01 vs. control, * p < 0.05, ** p < 0.01 vs. ox-LDL

Article Snippet: To analyze the effect of ox-LDL on HUVECs, the cells were treated with different concentrations (25, 50, 100, and 200 μg/ml) of ox-LDL for 24 h. In another experiment, HUVECs were pre-incubated with 100, 200, 400, and 800 pM of recombinant human Klotho (R&D Systems) protein for 1 h and subsequently treated with 50 μg/mlox-LDL for 24 h. These cells were then stained with 20 μl of 5 mg/ml MTT (Sigma-Aldrich) per well and incubated for 4 h at 37 °C.

Techniques: Microscopy, MTT Assay, Activity Assay, Incubation, Control

Journal: Lipids in Health and Disease

Article Title: Klotho ameliorates oxidized low density lipoprotein (ox-LDL)-induced oxidative stress via regulating LOX-1 and PI3K/Akt/eNOS pathways

doi: 10.1186/s12944-017-0447-0

Figure Lengend Snippet: Klotho inhibited ROS production induced by ox-LDL in HUVECs. HUVECs were pre-incubated with 200 pM of recombinant human Klotho for 1 h, then treated with ox-LDL (50 μg/mL) for another 24 h. Ox-LDL alone and Klotho alone were used as controls. a Images observed under an inverted fluorescence microscope. (Bar = 50 μm) ( b ) Output figure of the fluorescence intensity detected by flow cytometry. c Average fluorescence intensity: Mean = total area under the peak/the total number of cells. Data are shown as mean ± S.D. d Lipid peroxidation was assessed by measuring the MDA levels in HUVECs treated with ox-LDL and/or Klotho ( n = 3). Statistical differences are expressed as ## p < 0.01 vs. blank control; ** p < 0.01 vs. ox-LDL

Article Snippet: To analyze the effect of ox-LDL on HUVECs, the cells were treated with different concentrations (25, 50, 100, and 200 μg/ml) of ox-LDL for 24 h. In another experiment, HUVECs were pre-incubated with 100, 200, 400, and 800 pM of recombinant human Klotho (R&D Systems) protein for 1 h and subsequently treated with 50 μg/mlox-LDL for 24 h. These cells were then stained with 20 μl of 5 mg/ml MTT (Sigma-Aldrich) per well and incubated for 4 h at 37 °C.

Techniques: Incubation, Recombinant, Fluorescence, Microscopy, Flow Cytometry, Control

Journal: Lipids in Health and Disease

Article Title: Klotho ameliorates oxidized low density lipoprotein (ox-LDL)-induced oxidative stress via regulating LOX-1 and PI3K/Akt/eNOS pathways

doi: 10.1186/s12944-017-0447-0

Figure Lengend Snippet: SOD, Cu/Zn-SODandgp91 phox in ox-LDL and Klotho-treated HUVECs. Cellular treatment was the same as described in Fig. . a Intracellular SOD activity was determined by the hydroxylamine method. b , c and d Western blotanalysis of Cu/Zn-SOD and gp91 phoxexpression in pre-treated HUVECs. Densitometry of the probed bands from the western blot were analyzed by Image J2x. Values are means ± S.D. ( n = 3). Statistical differences are expressed as ## p < 0.01 vs. control; ** p < 0.01 vs. ox-LDL

Article Snippet: To analyze the effect of ox-LDL on HUVECs, the cells were treated with different concentrations (25, 50, 100, and 200 μg/ml) of ox-LDL for 24 h. In another experiment, HUVECs were pre-incubated with 100, 200, 400, and 800 pM of recombinant human Klotho (R&D Systems) protein for 1 h and subsequently treated with 50 μg/mlox-LDL for 24 h. These cells were then stained with 20 μl of 5 mg/ml MTT (Sigma-Aldrich) per well and incubated for 4 h at 37 °C.

Techniques: Activity Assay, Western Blot, Control

Journal: Lipids in Health and Disease

Article Title: Klotho ameliorates oxidized low density lipoprotein (ox-LDL)-induced oxidative stress via regulating LOX-1 and PI3K/Akt/eNOS pathways

doi: 10.1186/s12944-017-0447-0

Figure Lengend Snippet: Klotho regulated NO production in HUVECs. a NO production in pre-treated HUVECs. b , c , d , and e mRNA levels of iNOS, eNOS, PI3K, and Akt were measured by real time PCR. Values are means ± S.D. ( n = 3). Statistical differences are expressed as # p < 0.05 vs . control; * p < 0.05 vs . ox-LDL. ## p < 0.01 vs. control; ** p < 0.01 vs. ox-LDL

Article Snippet: To analyze the effect of ox-LDL on HUVECs, the cells were treated with different concentrations (25, 50, 100, and 200 μg/ml) of ox-LDL for 24 h. In another experiment, HUVECs were pre-incubated with 100, 200, 400, and 800 pM of recombinant human Klotho (R&D Systems) protein for 1 h and subsequently treated with 50 μg/mlox-LDL for 24 h. These cells were then stained with 20 μl of 5 mg/ml MTT (Sigma-Aldrich) per well and incubated for 4 h at 37 °C.

Techniques: Real-time Polymerase Chain Reaction, Control

Journal: Oncogene

Article Title: Therapeutic efficacy of a DNA vaccine targeting the endothelial tip cell antigen delta-like ligand 4 in mammary carcinoma.

doi: 10.1038/onc.2010.176

Figure Lengend Snippet: Figure 1 Immunization with plasmid-encoded DLL4 results in an immunological response in BALB/c mice. (a) Expression of human DLL4 was verified by transient transfection of HeLa cells with empty vector or the vector encoding DLL4 (pDLL4) and analyzed by western blot. Recombinant human DLL4 was used as a positive control (rhDLL4). (b) Female BALB/c mice were immunized either intramuscularly (IM) or intradermally (ID) according to the depicted protocols. All intradermal injections were given in combination with electroporation (EP). One week after the final immunization animals were challenged by orthotopic injection of D2F2/E2 or TUBO cells in the mammary fat pad. (c) To investigate if the vaccination gave rise to an immunological response, serum was taken 10 days after the final immunization (protocol 1) and analyzed by human DLL4 enzyme-linked immunosorbent assay (ELISA). The figure depicts a representative analysis for one out of four mice analyzed.

Article Snippet: Plates were coated overnight at 4 1C with 2 pmol of recombinant human DLL4 (Bioinvent International, Lund, Sweden) or 1 pmol of recombinant mouse DLL4 (R&D Systems) per well in 50ml of 0.1M sodium carbonate, pH 9.5.

Techniques: Plasmid Preparation, Expressing, Transfection, Western Blot, Recombinant, Positive Control, Electroporation, Injection, Enzyme-linked Immunosorbent Assay

Journal: Oncogene

Article Title: Therapeutic efficacy of a DNA vaccine targeting the endothelial tip cell antigen delta-like ligand 4 in mammary carcinoma.

doi: 10.1038/onc.2010.176

Figure Lengend Snippet: Figure 2 DNA vaccination against DLL4 results in suppression of tumor growth in BALB/c mice. (a) Expression of DLL4 in D2F2/ E2 tumor endothelial cells was assessed by immunofluorescent staining for the endothelial markers, CD31 (red) and DLL4 (green). Cell nuclei were counterstained by DAPI (blue). Growth of orthotopically implanted D2F2/E2 (b and c) or TUBO (d) mammary carcinomas in BALB/c mice following immunization with empty vector or with DLL4 plasmid DNA according to protocol 1 (3 ID þ EP) or protocol 2 (2 IM, 1 ID þ EP; n ¼ 8 for all groups). The data shown in (b and d) are representative of three independent experiments. *Po0.05, **Po0.01; Student’s t-test.

Article Snippet: Plates were coated overnight at 4 1C with 2 pmol of recombinant human DLL4 (Bioinvent International, Lund, Sweden) or 1 pmol of recombinant mouse DLL4 (R&D Systems) per well in 50ml of 0.1M sodium carbonate, pH 9.5.

Techniques: Expressing, Staining, Plasmid Preparation

Journal: Oncogene

Article Title: Therapeutic efficacy of a DNA vaccine targeting the endothelial tip cell antigen delta-like ligand 4 in mammary carcinoma.

doi: 10.1038/onc.2010.176

Figure Lengend Snippet: Figure 3 Targeting DLL4 by vaccination results in excessive formation of nonfunctional blood vessels. (a) Visual inspection of excised D2F2/E2 tumors revealed overt thrombosis in tumors from DLL4-vaccinated mice. (b) Blood vessel density of D2F2/E2 tumors visualized by immunostaining for the endothelial cell marker CD31 (red; n ¼ 5 for each group). (c) Perfused blood vessels of D2F2/E2 tumors as determined by vascular perfusion using fluorescein-labeled tomato lectin (green; n ¼ 5 for each group). The total area of the vascular bed was visualized by immunostaining for CD31 (red; n ¼ 5 for each group). The fraction perfused area was expressed as perfused area divided by total vascular area for each field. (d) Apoptotic index of D2F2/E2 tumor cells as determined by immunostaining for activated caspase-3 (n ¼ 5 for each group). *Po0.05, **Po0.01; Student’s t-test.

Article Snippet: Plates were coated overnight at 4 1C with 2 pmol of recombinant human DLL4 (Bioinvent International, Lund, Sweden) or 1 pmol of recombinant mouse DLL4 (R&D Systems) per well in 50ml of 0.1M sodium carbonate, pH 9.5.

Techniques: Immunostaining, Marker, Labeling

Journal: Oncogene

Article Title: Therapeutic efficacy of a DNA vaccine targeting the endothelial tip cell antigen delta-like ligand 4 in mammary carcinoma.

doi: 10.1038/onc.2010.176

Figure Lengend Snippet: Figure 4 T cells are not mediators of the effector functions of the DLL4 vaccine. (a) Mouse DLL4-specific T cell responses following immunization by protocol 1 (3 ID þ EP) or protocol 2 (2 IM, 1 ID þ EP) were assessed by IFN-g ELISpot by stimulation of splenocytes with mouse DLL4 peptides or recombinant mouse extracellular portion of DLL4 (n ¼ 5 for each group). Recombinant carcinoembryonic antigen protein and carcinoembryonic antigen peptides were used as negative controls. The polyclonal stimulant concanavalin A (conA) was used as a positive control. The experiment was repeated three times. (b) Splenocyte activity in mice following immunization by protocol 1 (3 ID þ EP) or protocol 2 (2 IM, 1 ID þ EP) at the time of sacrifice after tumor challenge was assessed by IFN-g ELISpot without antigen-specific stimulation (vector, n ¼ 9; protocol 1, n ¼ 8; protocol 2, n ¼ 5). (c) Depletion of CD4 þ or CD8 þ cells was initiated three days before tumor challenge by injection of anti-CD4 or anti-CD8 antibodies and verified by flow cytometric analysis of peripheral blood 5 days after injection of antibodies. Tumor growth curve from vector-immunized mice was duplicated from Figure 2c, which depicts an experiment performed simultaneously to the T cell depletion experiment. (d) Growth of orthotopically implanted D2F2/E2 tumors in control mice and in mice depleted of CD4 þ/CD8 þ cells following vaccination with DLL4 plasmid DNA (n ¼ 6 for all groups). **Po0.01, ***Po0.001; Student’s t-test.

Article Snippet: Plates were coated overnight at 4 1C with 2 pmol of recombinant human DLL4 (Bioinvent International, Lund, Sweden) or 1 pmol of recombinant mouse DLL4 (R&D Systems) per well in 50ml of 0.1M sodium carbonate, pH 9.5.

Techniques: Enzyme-linked Immunospot, Recombinant, Positive Control, Activity Assay, Plasmid Preparation, Injection, Control

Journal: Oncogene

Article Title: Therapeutic efficacy of a DNA vaccine targeting the endothelial tip cell antigen delta-like ligand 4 in mammary carcinoma.

doi: 10.1038/onc.2010.176

Figure Lengend Snippet: Figure 5 Tumor growth suppression in DLL4-vaccinated mice is mediated by induction of anti-DLL4 antibodies. (a) The presence of anti-mouse DLL4 antibodies in the serum of DLL4-immunized mice was analyzed by ELISA. The figure depicts a representative analysis for one out of four mice analyzed. (b) IsolectinB4 staining of wholemounted P5 mouse retinas from mice treated with immune serum from vector or DLL4 immunized mice according to protocol 1 (3 ID þ EP; n ¼ 4 for each group). Arrowheads indicate endothelial cells morphologically identified as tip cells. (c) Growth of orthotopically implanted D2F2/E2 tumors in mice treated with serum from mice immunized with vector (n ¼ 12) or DLL4 plasmid DNA (n ¼ 16) according to protocol 1 (3 ID þ EP). The experiment was repeated twice with similar results. **Po0.01; Student’s t-test.

Article Snippet: Plates were coated overnight at 4 1C with 2 pmol of recombinant human DLL4 (Bioinvent International, Lund, Sweden) or 1 pmol of recombinant mouse DLL4 (R&D Systems) per well in 50ml of 0.1M sodium carbonate, pH 9.5.

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Plasmid Preparation

Journal: Oncogene

Article Title: Therapeutic efficacy of a DNA vaccine targeting the endothelial tip cell antigen delta-like ligand 4 in mammary carcinoma.

doi: 10.1038/onc.2010.176

Figure Lengend Snippet: Figure 6 No evidence for liver toxicity or delayed wound healing following immunization with the DLL4 vaccine. (a) Hematoxylin and eosin staining of tissue sections of liver from mice that initiated the immunization procedure with empty vector or the DLL4 vaccine 5 months earlier. (b) Rate of healing expressed as % closure of wounds inflicted to mice immunized with empty vector or the DLL4 vaccine (n ¼ 8 mice per group, two wounds per mouse). *Po0.05, repeated-measures ANOVA.

Article Snippet: Plates were coated overnight at 4 1C with 2 pmol of recombinant human DLL4 (Bioinvent International, Lund, Sweden) or 1 pmol of recombinant mouse DLL4 (R&D Systems) per well in 50ml of 0.1M sodium carbonate, pH 9.5.

Techniques: Staining, Plasmid Preparation

Journal: Frontiers in Immunology

Article Title: Differential Expression of miRNAs in Trichloroethene-Mediated Inflammatory/Autoimmune Response and Its Modulation by Sulforaphane: Delineating the Role of miRNA-21 and miRNA-690

doi: 10.3389/fimmu.2022.868539

Figure Lengend Snippet: DCAC treatment led to increased miR-21 expression in RAW 264.7 cells. (A) Increased expression of miR-21 in the DCAC-treated RAW 264.7 cells and its amelioration by SFN. (B, C) mRNA expression of NF-κB (p65) and IL-12 in RAW 264.7 cells transfected with miR-21 inhibitors. n = 4 in each group. (D) Protein expression of NF-κB (p65) and its densitometry analysis in RAW 264.7 cells following DCAC treatment and transfection with miR-21 inhibitors. n = 3 in each group. *p < 0.05 vs. controls; # p < 0.05 vs. DCAC-treated RAW 264.7 cells. DCAC, dichloroacetyl chloride; SFN, sulforaphane.

Article Snippet: RAW 264.7 cells (30,000 cells/100 μl in BPS medium) were plated in 96-well plates and allowed to adhere for 24 h. For transfection, 100 ng of DNA constructs [NF-κB Transient Pack (BPS Biosciences, San Diego, CA, USA) consisting of NF-κB reporter vector + constitutively expressing Renilla luciferase vector or NC reporter (non-inducible luciferase vector + Renilla luciferase vector)] along with 20 nM of miR-690 mimics or respective 20 nM of miRNA control were co-transfected into RAW 264.7 cells using Lipofectamine 2000 (Thermo Fisher Scientific).

Techniques: Expressing, Transfection

Journal: Frontiers in Immunology

Article Title: Differential Expression of miRNAs in Trichloroethene-Mediated Inflammatory/Autoimmune Response and Its Modulation by Sulforaphane: Delineating the Role of miRNA-21 and miRNA-690

doi: 10.3389/fimmu.2022.868539

Figure Lengend Snippet: DCAC treatment led to increased expression of miR-690 in RAW 264.7 cells. (A) Increased expression of miR-690 in the DCAC-treated RAW 264.7 cells. SFN treatment ameliorated the effect of DCAC. (B, C) mRNA expression of NF-κB (p65) and IL-12 in RAW 264.7 cells transfected with miR-690 mimics. n = 4 in each group. (D) Protein expression of NF-κB (p65) and its densitometry analysis in RAW 264.7 cells. (E) NF-κB-luciferase reporter plasmid along with miR-690 mimics or control were co-transfected into RAW 264.7 cells. (F) Predicted miR-690 targeting region in 3′ UTR of NFkB. Luciferase activity is the ratio of Renilla/ Firefly . Renilla is raw renilla luciferase activity, whereas Firefly is firefly luciferase activity. The 3′ UTR starts at bp 6,577 and ends at bp 6,627. n = 3 in each group. *p < 0.05 vs. controls; # p < 0.05 vs. DCAC-treated RAW 264.7 cells. DCAC, dichloroacetyl chloride; SFN, sulforaphane.

Article Snippet: RAW 264.7 cells (30,000 cells/100 μl in BPS medium) were plated in 96-well plates and allowed to adhere for 24 h. For transfection, 100 ng of DNA constructs [NF-κB Transient Pack (BPS Biosciences, San Diego, CA, USA) consisting of NF-κB reporter vector + constitutively expressing Renilla luciferase vector or NC reporter (non-inducible luciferase vector + Renilla luciferase vector)] along with 20 nM of miR-690 mimics or respective 20 nM of miRNA control were co-transfected into RAW 264.7 cells using Lipofectamine 2000 (Thermo Fisher Scientific).

Techniques: Expressing, Transfection, Luciferase, Plasmid Preparation, Activity Assay

Journal: Cell

Article Title: Broad neutralization of SARS-CoV-2 variants by an inhalable bispecific single-domain antibody

doi: 10.1016/j.cell.2022.03.009

Figure Lengend Snippet:

Article Snippet: SARS-CoV-2 Spike RBD-His (Alpha) , , Sino Biological Inc. , Cat# 40592-V08H18.

Techniques: Produced, Virus, Recombinant, Saline, Modification, Luciferase, Antibody Labeling, Clone Assay, Expressing, Plasmid Preparation, Software

Journal: Vaccine

Article Title: A cell-based ELISA as surrogate of virus neutralization assay for RBD SARS-CoV-2 specific antibodies

doi: 10.1016/j.vaccine.2022.02.044

Figure Lengend Snippet: Detection of SARS-CoV-2 NAbs by a cell based ELISA-virus neutralization test (cbE-VNT). The illustrations were created using BioRender.

Article Snippet: Next, 50 μl of human Fc-RBD (RBD-hFc) or mouse Fc-RBD (RBD-mFc) fusion proteins, or SARS-CoV-2 Spike S1 (Sino-Biological-Inc #40591-V08H3) at different concentrations were added and incubated for 2 h at RT.

Techniques: In-Cell ELISA, Neutralization

Journal: ACS Infectious Diseases

Article Title: Microfluidic Antibody Affinity Profiling Reveals the Role of Memory Reactivation and Cross-Reactivity in the Defense Against SARS-CoV-2

doi: 10.1021/acsinfecdis.1c00486

Figure Lengend Snippet: Microfluidic antibody affinity profiling (MAAP) of nine convalescent COVID-19 patients and three pre-pandemic sera. (A) Probability density plots of MAAP against fluorescently labeled SARS-CoV-2 RBD, spike S1, and spike S2. The graphs show the affinity ( K D ) and the molar concentration of antibody binding sites for each convalescent serum sample. No binding could be detected in the three pre-pandemic sera. Points correspond to the maximum a posteriori values in the two-dimensional posterior probability distribution, and shaded regions correspond to the probability density. Gray shaded regions indicate the area of nonbinding for samples with [antibody binding sites] < K D . (B) ELISA −log (EC 50 ) values for the same nine convalescent COVID-19 patient samples and three pre-pandemic sera ( 1 : NL63+, 229E+; 2 : NL63+, 229E+, OC43+, HKU1+; 3 : 229E+). A SARS-CoV-2 seronegative sample and a SARS-CoV-2 seropositive sample served as negative and positive controls, respectively. Immobilized antigens are SARS-CoV-2 spike ectodomain, spike S1, and the RBD. Detection was performed with fluorescently labeled anti-human IgG antibodies. (A, B) Red boxes/circles indicate convalescent sera 4 and 5 , which have the strongest immune response to all SARS-CoV-2 spike subunits.

Article Snippet: HCoV-NL63 Spike RBD (10605-CV) and HCoV-HKU1 Spike RBD (10600-CV) were obtained from R&D Systems.

Techniques: Labeling, Concentration Assay, Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: ACS Infectious Diseases

Article Title: Microfluidic Antibody Affinity Profiling Reveals the Role of Memory Reactivation and Cross-Reactivity in the Defense Against SARS-CoV-2

doi: 10.1021/acsinfecdis.1c00486

Figure Lengend Snippet: Microfluidic antibody affinity profiling against HCoV-NL63 spike S1 and RBD to establish cross-reactivity of antibodies in convalescent COVID-19 serum. (A, B) Equilibrium binding curves of a neutralizing SARS-CoV-2 antibody against 10 nM fluorescently labeled spike S1 from (A) HCoV-HKU1 or (B) HCoV-NL63, respectively. The hydrodynamic radii ( R h ) of the free labeled spike proteins did not increase upon addition of the antibody, indicating the absence of binding. (C) Equilibrium binding curve of an anti-HKU1 antibody against 10 nM fluorescently labeled spike S1 of HCoV-HKU1 showed very tight binding with a K D below 0.1 nM. The K D was determined by nonlinear least-squares fitting using eq . (D) Probability density plots of MAAP against fluorescently labeled HCoV-NL63 spike S1 and HCoV-NL63 RBD. The graphs show the affinity ( K D ) and the molar concentration of antibody binding sites for each of the convalescent COVID-19 (red) and the pre-pandemic sera (blue), whereby pre-pandemic sera and 2 were found to be seropositive for HCoV-NL63. Points correspond to the maximum a posteriori values in the two-dimensional posterior probability distribution, and shaded regions correspond to the probability density.

Article Snippet: HCoV-NL63 Spike RBD (10605-CV) and HCoV-HKU1 Spike RBD (10600-CV) were obtained from R&D Systems.

Techniques: Binding Assay, Labeling, Concentration Assay

Journal: ACS Infectious Diseases

Article Title: Microfluidic Antibody Affinity Profiling Reveals the Role of Memory Reactivation and Cross-Reactivity in the Defense Against SARS-CoV-2

doi: 10.1021/acsinfecdis.1c00486

Figure Lengend Snippet: (A) Schematic of SARS-CoV-2–HCoV-NL63 cross-reactivity competition assay. Typically, 10 nM fluorescently labeled HCoV-NL63 RBD was mixed either with buffer or with patient serum, incubated for 1 h, and subjected to microfluidic diffusional sizing to determine the size of the free labeled RBD ( R h,free ) and the size of the immune-complex ( R h,complex ). For binding competition, 250 nM unlabeled SARS-CoV-2 RBD was added to the mixture of 10 nM HCoV-NL63 RBD and patient serum and incubated for 1 h before measuring the hydrodynamic radius ( R h ). If the serum contains cross-reactive antibodies, a size decrease is observed, as the antibodies will bind to the excess of unlabeled RBD (left box). If the antibodies in the serum are not cross-reactive, they remain bound to the labeled HCoV-NL63 RBD, and the R h of the immuno-complex will stay constant (right box). (B) SARS-CoV-2–HCoV-NL63 cross-reactivity competition assay in convalescent COVID-19 sera and pre-pandemic sera. Filled circles (●) indicate the size of the HCoV-NL63 RBD–antibody immune complex. Empty circles (○) indicate the size of HCoV-NL63 RBD after addition of excess unlabeled SARS-CoV-2 RBD. (Red) Convalescent COVID-19 sera, (Blue) pre-pandemic sera. No decrease of R h upon addition of the unlabeled SARS-CoV-2 RBD could be observed in any of the sera, indicating no cross-reactivity of anti-NL63 antibodies with SARS-CoV-2 RBD.

Article Snippet: HCoV-NL63 Spike RBD (10605-CV) and HCoV-HKU1 Spike RBD (10600-CV) were obtained from R&D Systems.

Techniques: Competitive Binding Assay, Labeling, Incubation, Binding Assay