spike protein subunit 1  (RayBiotech inc)

Journal: medRxiv

Article Title: A Phase II Study Integrating a Single-Blind Safety Phase with a Double-Blind, Placebo-Controlled Randomized Phase, Assessing Single-Dose Intramuscular or Intranasal Administration to Evaluate the Safety and Immunogenicity of the Recombinant Vaccine Against COVID-19 (AVX/COVID-12 “Patria”) Based on an Active Newcastle Disease Viral Vector as a Heterologous Booster in Subjects with Evidence of Previous Immunity to SARS-CoV-2

doi: 10.1101/2024.02.11.24302594

Figure Lengend Snippet:

Article Snippet: Duplicate samples of 2 x 10 5 cells/well were plated in 96-well plates and stimulated with 5 µg/mL of the spike protein subunit 1 (RayBiotech, Peachtree Corners, GA), using culture medium as a negative control.

Techniques:

Journal: Frontiers in Immunology

Article Title: Robust memory humoral immune response to SARS-CoV-2 in the tonsils of adults and children

doi: 10.3389/fimmu.2023.1291534

Figure Lengend Snippet: Tonsils processing and stimulation. (A) SARS-CoV-2 positive samples. Nasopharyngeal and oropharyngeal swabs were collected from each patient in viral transport media (VTM). Six laboratory-confirmed RT-PCR SARS-CoV-2 positive samples with Ct values of 18-25 were selected and pooled into one tube. The pooled samples were mixed well, aliquoted, and stored at -80°C to ensure that the same stimulant (viral load) was used for each tonsillar sample. (B) TMNCs isolation. Fresh tonsillar samples were processed within one hour after surgery. The tissues were cut into small pieces using a sterilized scalpel and then checked grossly. Then, the cell suspension was passed through a 70-μm sterile nylon mesh. TMNCs were separated by density gradient centrifugation using Ficoll-Paque (400 x g for 30 min). The cells were then washed twice using sterile phosphate-buffered saline (PBS) and resuspended in 5 ml complete RPMI 1640 medium for further processing and cell culture steps. (C) TMNCs stimulation. One aliquot of stored stimulation sample was thawed and 10 μl were added to 4 × 10 6 cells/ml of TMNC suspension from each individual in 96 well plates. Negative control samples (unstimulated) were prepared by adding 10 μl of VTM only to a TMNC suspension from each individual. Following gentle mixing, 96-well cell culture plates were incubated in a 5% CO 2 incubator at 37°C. Cell culture supernatants were collected after 10 days and stored at −80°C until assayed.

Article Snippet: Recombinant SARS-CoV-2 S1 subunit (amino acids 1–685), RBD (amino acids 319–541), and full-length nucleocapsid (N) proteins were purchased commercially (Sino Biological, China).

Techniques: Reverse Transcription Polymerase Chain Reaction, Isolation, Suspension, Sterility, Gradient Centrifugation, Saline, Cell Culture, Negative Control, Gentle, Incubation

Journal: Frontiers in Immunology

Article Title: Robust memory humoral immune response to SARS-CoV-2 in the tonsils of adults and children

doi: 10.3389/fimmu.2023.1291534

Figure Lengend Snippet: Serum antibody response. (A) Endpoint binding IgG titers in serum samples from adults and children were determined against SARS-CoV-2 N, S1, and RBD proteins. The dotted lines represent the cutoff of each assay. (B) Inhibitory activity of serum samples from adults and children against ancestral Wuhan strain and Omicron BA.1 variant pseudoviruses were determined. (C) Heatmap showing neutralizing antibodies IC 50 obtained from analysis of panel (B) against ancestral Wuhan strain and Omicron BA.1 variant pseudoviruses. Fold change in IC 50 is shown on top of the heatmap representing activity against the SARS-CoV-2 Omicron BA.1 variant relative to the ancestral Wuhan SARS-CoV-2 strain. (D) Correlation between neutralizing antibodies against the SARS-CoV-2 Omicron BA.1 variant and the ancestral Wuhan SARS-CoV-2 strain and binding IgG antibodies in serum is shown for anti-RBD, -S1, and -N antibodies from adults and children.

Article Snippet: Recombinant SARS-CoV-2 S1 subunit (amino acids 1–685), RBD (amino acids 319–541), and full-length nucleocapsid (N) proteins were purchased commercially (Sino Biological, China).

Techniques: Binding Assay, Activity Assay, Variant Assay

Journal: Frontiers in Immunology

Article Title: Robust memory humoral immune response to SARS-CoV-2 in the tonsils of adults and children

doi: 10.3389/fimmu.2023.1291534

Figure Lengend Snippet: Induced antibodies in stimulated tonsils supernatants. Supernatants from stimulated or unstimulated mononuclear cells from tonsils were tested for binding and neutralizing antibodies. (A) Endpoint binding IgG titers in supernatants (unstimulated and stimulated) from adults and children were determined against SARS-CoV-2 N, S1, and RBD proteins. The mean titer is shown on top of each bar. (B) The inhibitory activity in supernatants (unstimulated and stimulated) from adults and children against ancestral Wuhan and Omicron BA.1 pseudoviruses was determined. Heatmap shows neutralizing antibodies (IC 50 ) obtained from unstimulated and stimulated against ancestral Wuhan and Omicron BA.1 pseudoviruses. (C) Correlation between neutralizing antibody against the SARS-CoV-2 Omicron BA.1 variant and the ancestral Wuhan SARS-CoV-2 strain and binding IgG antibodies in the supernatants (unstimulated and stimulated) is shown from anti-RBD, -S1 and -N antibodies from adults and children. (D) Correlation between neutralizing antibody titers against the ancestral Wuhan SARS-CoV-2 strain vs the SARS-CoV-2 Omicron BA.1 variant is shown.

Article Snippet: Recombinant SARS-CoV-2 S1 subunit (amino acids 1–685), RBD (amino acids 319–541), and full-length nucleocapsid (N) proteins were purchased commercially (Sino Biological, China).

Techniques: Binding Assay, Activity Assay, Variant Assay