v08b b 229e cov spike sino biological cat  (Sino Biological)

Journal: Cell Host & Microbe

Article Title: Rare, convergent antibodies targeting the stem helix broadly neutralize diverse betacoronaviruses

doi: 10.1016/j.chom.2022.10.010

Figure Lengend Snippet: Identification of broadly neutralizing antibodies against human betacoronaviruses The area under the curve (AUC) from titration of mAb binding to spike proteins from human betacoronaviruses SARS-CoV-2 Wuhan-Hu-1 (CoV-2), SARS-CoV (CoV-1), MERS-CoV, HCoV-HKU1, and HCoV-OC43, as well as alphacoronaviruses HCoV-NL63 and HCoV-229E, is shown on the panel of the left. Influenza H1 hemagglutinin (HA) was included as a control antigen and L9 IgG1 (malaria specific; Wang et al., 2020 ) was included as a negative control mAb for binding experiments. AUC values for each antigen are shown after subtraction with values for the negative control antigen CD4. The antibody titers at 50% neutralization (NT 50 ) against SARS-CoV-2 Wuhan-Hu-1, SARS-CoV, MERS-CoV, HCoV-NL63 envelope-pseudotyped virus, as well as authentic HCoV-OC43, are shown on the right. Neutralizing mAbs are ranked by their breadth of neutralization and the geometric mean of their NT 50 values. Cells highlighted in blue denote mAbs that did not show neutralizing potency at the highest concentration tested (100 μg/mL). Negative control mAbs for neutralization are DEN3 (dengue-specific; Rogers et al., 2020 ) for SARS-CoV-2, SARS-CoV, MERS-CoV and HCoV-NL63, and CV503 (SARS-CoV-2 RBD-specific; Cho et al., 2021 ) for HCoV-OC43. NT 50 values were calculated using the dose-response-inhibition model with 5-parameter Hill slope equation in GraphPad Prism version 9.3.1.

Article Snippet: 229E-CoV spike , Sino Biological , Cat. # 40605-V08B-B.

Techniques: Titration, Binding Assay, Negative Control, Neutralization, Concentration Assay, Inhibition

Journal: Cell Host & Microbe

Article Title: Rare, convergent antibodies targeting the stem helix broadly neutralize diverse betacoronaviruses

doi: 10.1016/j.chom.2022.10.010

Figure Lengend Snippet:

Article Snippet: 229E-CoV spike , Sino Biological , Cat. # 40605-V08B-B.

Techniques: Recombinant, Transfection, Lysis, Cell Culture, Luciferase, Expressing, Irradiation, Amplification, Plasmid Preparation, Software, Fluorescence, Microscopy, Imaging

Journal: Scientific Reports

Article Title: Cross-reactive antibodies elicited to conserved epitopes on SARS-CoV-2 spike protein after infection and vaccination

doi: 10.1038/s41598-022-10230-y

Figure Lengend Snippet: Vaccination targeting SARS-CoV-2 did not significantly affect antibody responses against most seasonal coronaviruses. ( A ) Bar and line graph of antibody responses obtained by ELISA at different plasma dilutions to Human CoV OC43 spike protein, before the Pfizer-BioNTech BNT162b2 vaccine was administered (Week 0) and after both doses (Week 7), using plasma from patients with no history of SARS-CoV-2 infection, n = 24 individuals, p ≤ 0.0001. ( B ) Bar and line graph of antibody responses obtained by ELISA at different plasma dilutions to Human CoV HKU1 spike protein, before the Pfizer-BioNTech BNT162b2 vaccine was administered (Week 0) and after both doses (Week 7), using plasma from patients with no history of SARS-CoV-2 infection, n = 24 individuals, p ≥ 0.99. ( C ) Bar and line graph of antibody responses obtained by ELISA at different plasma dilutions to Human CoV 229E spike protein, before the Pfizer-BioNTech BNT162b2 vaccine was administered (Week 0) and after both doses (Week 7), using plasma from patients with no history of SARS-CoV-2 infection, n = 24 individuals, p = 0.75. ( D ) Bar and line graph of antibody responses obtained by ELISA at different plasma dilutions to Human CoV NL63 spike protein, before the Pfizer-BioNTech BNT162b2 vaccine was administered (Week 0) and after both doses (Week 7), using plasma from patients with no history of SARS-CoV-2 infection, n = 24 individuals, p ≤ 0.0001. Endpoint titers are designated by the most dilute plasma concentration detected above the minimum threshold of the background OD450 multiplied by 3. Dots are representative of individual samples. Bars are representative of the mean of all samples. Statistical analysis was performed using Wilcoxon matched-pairs signed rank test with two-tailed P values reported.

Article Snippet: ELISAs were performed using the following antigens: SARS-CoV-1 (Cat# 10683-CV, Lot# DOXT0120121, R&D Systems, Minneapolis, MN, USA), SARS-CoV-2 (Cat# 10549-CV, Lot# DODR0221011, R&D Systems), Bat CoV RaTG13 (Cat# 10660-CV, Lot# DOWW0120121, R&D Systems), MERS-CoV spike protein (Cat# 40069-V08B, Lot# LC14AP2305, Sino Biological, Wayne, PA, USA), Human CoV NL63 spike protein (Cat# 40600-V08H, Lot#LC14NO2607, Sino Biological), Human CoV 229E spike protein (Cat# 40605-V08H, Lot# LC14AP2302, Sino Biological), Human CoV HKU1 spike protein (Cat# 40021-V08H, Lot# LC14AP2707, Sino Biological), Human CoV OC43 spike protein (Cat# 40607-V08H, Lot#LC14DE1609, Sino Biological) were all diluted to 1 ug/mL in 0.1 M sodium bicarbonate and incubated on high-binding plates (3369, Corning Inc, Corning, NY, USA) overnight at 4 degrees.

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Concentration Assay, Two Tailed Test

Journal: Journal of Immunological Methods

Article Title: Highly versatile antibody binding assay for the detection of SARS-CoV-2 infection and vaccination

doi: 10.1016/j.jim.2021.113165

Figure Lengend Snippet: Time course of key biomarkers in SARS-CoV-2 infection, adapted from BioRender.com . The solid green line represents a typical trajectory of the RT-PCR data for viral nucleic acid from respiratory samples, while the broken purple line indicates a typical virus-specific antibody trajectory in peripheral blood, relative to time of infection, as indicated. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Spike Protein S1 from non-SARS-CoV-2 coronaviruses (HCoV-229E, HCoV-NL63, HCoV-HKU-1) and Spike Protein S1 and S2 extracellular domain (HCoV-OC43) were obtained from Sino Biologicals (Wayne, PA, USA) and pooled in equimolar amounts to a final concentration of 1 mg/ml.

Techniques: Infection, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of Immunological Methods

Article Title: Highly versatile antibody binding assay for the detection of SARS-CoV-2 infection and vaccination

doi: 10.1016/j.jim.2021.113165

Figure Lengend Snippet: S1 RBD antibody binding assay. (A) Violin plots showing IgG reactivity of pre-COVID-19 sera ( n = 19) to RBD (blue) and a pool of non-SARS-CoV-2 coronavirus antigens (red) determined by ELISA using sera diluted 1:20. (B) Comparison among four serological assays. The dot plots show IgG results obtained with our in-house automated ELISA (RBD-based) (pink symbols), Roche Elecsys® Anti-SARS-CoV-2 (N-based) (green symbols), Abbott Architect SARS-CoV-2 IgG assay (N-based) (blue symbols), and the Roche Elecsys® Anti-SARS-CoV-2 S (S-based) (brown symbols). Serum/plasma samples were obtained from subjects who tested PCR-positive for SARS-CoV-2 infection (open circles, n = 30); pre-COVID-19 samples (open triangles, n = 70 for all assays except the Roche spike-based assay for which we used a subset of n = 57). Assays were performed in technical duplicates. Each symbol represents one study subject. Black horizontal lines represent cut-off values each serological assay. Cut-off values for the commercial assays were as per manufacturer's instructions. For the in-house ELISA, the cut-off value (OD 405 = 0.3) was calculated as the mean + 3 SD obtained with sera from 103 SARS-CoV-2 PCR-negative subjects who remained negative for at least 16 weeks after the blood draw utilized in the assay (data shown in ). It is noted that, for clarity purposes, a single scale (Index on the right y axis) was used for both commercial assays. However, the index calculation is different in the two assays; therefore, the relative numbers cannot be compared across assays. CI, confidence interval. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Spike Protein S1 from non-SARS-CoV-2 coronaviruses (HCoV-229E, HCoV-NL63, HCoV-HKU-1) and Spike Protein S1 and S2 extracellular domain (HCoV-OC43) were obtained from Sino Biologicals (Wayne, PA, USA) and pooled in equimolar amounts to a final concentration of 1 mg/ml.

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

Journal: Journal of Immunological Methods

Article Title: Highly versatile antibody binding assay for the detection of SARS-CoV-2 infection and vaccination

doi: 10.1016/j.jim.2021.113165

Figure Lengend Snippet: Applications of the in-house S1 RBD antibody binding assay. Violin plots showing the reactivity to RBD of serum/plasma samples (diluted 1:80) from convalescent subjects who had tested PCR-positive for SARS-CoV-2 infection ( n = 83, blue); COVID-19 hospitalized subjects ( n = 146, red); subjects who received full COVID-19 vaccination ( n = 283, green); adult residents in the Lakewood, NJ township (high burden) s( n = 148, purple); pre-COVID-19 samples ( n = 104; orange); SARS-CoV-2 PCR-negative subjects ( n = 103; black). In all panels, the solid horizontal lines represent the median (thick line) and interquartile range (thin lines). Assays were performed in technical duplicates. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Spike Protein S1 from non-SARS-CoV-2 coronaviruses (HCoV-229E, HCoV-NL63, HCoV-HKU-1) and Spike Protein S1 and S2 extracellular domain (HCoV-OC43) were obtained from Sino Biologicals (Wayne, PA, USA) and pooled in equimolar amounts to a final concentration of 1 mg/ml.

Techniques: Binding Assay, Infection

Journal: Journal of Immunological Methods

Article Title: Highly versatile antibody binding assay for the detection of SARS-CoV-2 infection and vaccination

doi: 10.1016/j.jim.2021.113165

Figure Lengend Snippet: Comparison of dried blood microsampling and phlebotomy for blood collection. (A) Dried-blood microsampling. The dot plot shows RBD-specific IgG antibody binding determined by ELISA with samples collected by finger stick utilizing Mitra microsamplers from subjects that tested PCR-negative ( n = 50, circles) or PCR-positive for SARS-CoV-2 infection ( n = 12, triangles). The horizontal line represents the cut-off value of the assay, calculated as in the legend to . (B) Correlation between ELISA results obtained from the same subjects by phlebotomy and by finger stick and Mitra microsampling. The correlation plot shows results obtained with samples from 16 subjects (13 seropositive and 3 seronegative for SARS-CoV-2 infection). Assays were performed in technical duplicates. The Pearson correlation coefficient (R 2 ) of the comparison is also shown. In both panels, each symbol represents one study subject.

Article Snippet: Spike Protein S1 from non-SARS-CoV-2 coronaviruses (HCoV-229E, HCoV-NL63, HCoV-HKU-1) and Spike Protein S1 and S2 extracellular domain (HCoV-OC43) were obtained from Sino Biologicals (Wayne, PA, USA) and pooled in equimolar amounts to a final concentration of 1 mg/ml.

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

Journal: Journal of Immunological Methods

Article Title: Highly versatile antibody binding assay for the detection of SARS-CoV-2 infection and vaccination

doi: 10.1016/j.jim.2021.113165

Figure Lengend Snippet: Matrix equivalency assays. The dot plot shows RBD-specific IgG antibody binding determined by ELISA. (A) Serum and plasma matrices. Serum and plasma samples were obtained from SARS-CoV-2 PCR-negative subjects (n = 5) using concurrently five different collection tubes, as indicated by the different symbols. (B) Breast milk matrix. Breast milk samples were collected from four SARS-CoV-2 PCR-negative lactating mothers. (A,B) All matrices (serum and plasma in panel A and breast milk in panel B) were used to serially dilute serum from one SARS-CoV-2 antibody positive subject at 1:20, 1:80, and 1:320, as indicated. The same seropositive sample was also diluted in conventional ELISA blocking buffer (reference condition, black circle). No serum indicates ELISA results obtained with the various matrices only (serum, plasma, breast milk), in the absence of the diluted seropositive serum. Assays were performed in technical duplicates. In both panels, each symbol represents one matrix per study subject.

Article Snippet: Spike Protein S1 from non-SARS-CoV-2 coronaviruses (HCoV-229E, HCoV-NL63, HCoV-HKU-1) and Spike Protein S1 and S2 extracellular domain (HCoV-OC43) were obtained from Sino Biologicals (Wayne, PA, USA) and pooled in equimolar amounts to a final concentration of 1 mg/ml.

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