Journal: Cell Reports

Article Title: A two-adjuvant multiantigen candidate vaccine induces superior protective immune responses against SARS-CoV-2 challenge

doi: 10.1016/j.celrep.2021.110112

Figure Lengend Snippet: Neutralization of SARS-CoV-2 pseudovirus in sera and detection of N- and S-specific T cell responses against the B.1351 and B.1.617.2 variants SARS-CoV-2 wild-type (WT), B.1.351 variant, or B.1.617 variant pseudoviruses were incubated with different serum sample dilutions for 1 h at 37°C before the mixtures were added to ACE2-overexpressing 293T cells. Transduction efficiency was quantified by measuring virus-encoded luciferase activity in cell lysates 48 h after transduction and used to calculate the serum dilution factor that resulted in a 50% reduction in pseudovirus particles that were associated with different degrees of S protein-mediated cell entry. (A and B) The 50% pseudovirus neutralization (pVNT 50 ) in serum from mice immunized with the candidate vaccine (A) and mice immunized with the inactivated vaccine (B) against the B.1.351 and B.1.617.2 variants compared with that against the wild-type (WT) virus (n = 4 mice per group). (C–F) Antigen-specific activation of T cells by the N and S proteins of the B.1.351 and B.1.617.2 variants compared with the homologous WT proteins. N-specific (C) and S-specific (D) activation of T cells in splenocytes from mice immunized with the candidate vaccine or the inactivated vaccine at 14 days after the third immunization. N-specific (E) and S-specific (F) activation of T cells in lung tissues from mice immunized with the candidate vaccine or the inactivated vaccine at 14 days after the third immunization (n = 4 mice per group). Significance was determined via one-way ANOVA with a Tukey multiple comparison test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ns, no significance.

Article Snippet: B.1.351 variant N protein , SinoBiological , Cat# 40588-V07E9.

Techniques: Neutralization, Variant Assay, Incubation, Transduction, Luciferase, Activity Assay, Activation Assay

Journal: Cell Reports

Article Title: A two-adjuvant multiantigen candidate vaccine induces superior protective immune responses against SARS-CoV-2 challenge

doi: 10.1016/j.celrep.2021.110112

Figure Lengend Snippet:

Article Snippet: B.1.351 variant N protein , SinoBiological , Cat# 40588-V07E9.

Techniques: Blocking Assay, Recombinant, Variant Assay, Enzyme-linked Immunospot, Luciferase, Reporter Assay, Enzyme-linked Immunosorbent Assay, Transgenic Assay, Software, Filtration, Modification

Journal: Analytical Chemistry

Article Title: Development of a Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of SARS-CoV-2 Spike Glycoprotein and Nucleoprotein

doi: 10.1021/acs.analchem.0c02288

Figure Lengend Snippet: Figure 2. Schematic of the workflow used to develop a PRM assay for the detection and quantitation of SARS-CoV-2 spike and nucleoprotein A) PRM assay development was performed using recombinant SARS CoV-2 spike protein and nucleoprotein. Proteotypic target peptides/transitions were selected to generate a spectral library in Skyline. B) The PRM assay was then used to quantitate the SARS CoV-2 protein levels in a mock sample that was created by adding an inactivated virus sample to in-vitro derived mucus.

Article Snippet: SARS-CoV-2 nucleocapsid-His recombinant protein was purchased from Sino Biological (100 g, 40588-V08B,) The protein was difficult to dissolve and required treatment before S-trap preparation below with 40L dimethyl sulfoxide (276855-100mL, Sigma), 126 L 1% TFA, and 100L 1x S-Trap lysis buffer (5% SDS, 50 mM TEAB, pH adjusted to 7.55 using 12% phosphoric acid).

Techniques: Quantitation Assay, Recombinant, Targeted Proteomics, Virus, In Vitro, Derivative Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: Aptamers targeting SARS-CoV-2 nucleocapsid protein exhibit potential anti pan-coronavirus activity

doi: 10.1038/s41392-024-01748-w

Figure Lengend Snippet: Prediction of binding and affinity identification between the aptamers and the N proteins of SARS, MERS and -OC43. a Prediction of the binding affinity between the aptamers and N proteins of SARS, MERS and HCoV-OC43 via Zdock score. The N protein of SARS-CoV-2 and ALB protein were used as positive and negative control, respectively. The characterizations of affinity and specificity between aptamers and N proteins of ( b ) SARS, ( c ) MERS and ( d ) -OC43 were performed via CE. e The results of molecular docking to predict and simulate the binding sites of aptamer Seq-1022 to N proteins of SARS, MERS and -OC43 using Discovery Studio software

Article Snippet: SARS NP (40143-V08B), MERS NP (40068-V08B), RBD of spike protein of SARS-CoV-2 Omicron strain (40592-V08H121) and SARS-CoV-2 Nucleocapsid monoclonal antibody (40588-R0004) were purchased from Sino Biological Inc. Normal human serum was obtained from Beijing BioDee Biotechnology Co., Ltd. Human albumin was obtained from Sigma-Aldrich (St. Louis, MO).

Techniques: Binding Assay, Negative Control, Software

Journal: Vaccine

Article Title: The recombinant N-terminal domain of spike proteins is a potential vaccine against Middle East respiratory syndrome coronavirus (MERS-CoV) infection

doi: 10.1016/j.vaccine.2016.11.064

Figure Lengend Snippet: Description of the N-terminal domain (NTD) immunogen and vaccination schedule. (A) The location of the NTD protein on the Middle East respiratory syndrome coronavirus MERS-CoV spike (S) protein. The recombinant (r)NTD protein consists of 336 amino acid (aa) residues (18–353) of S protein. A gp67 signal peptide (SP) was added to the N terminus for expression of the rNTD protein. (B) Purified rNTD protein detected by SDS-PAGE (left) and Western blot (right). The purified rNTD protein was separated by a 10% SDS-PAGE and stained with 0.25% Coomassie brilliant blue. Anti-NTD polyclonal antibody and infrared ray-labeled secondary antibody were used for the Western blot assay. Lane 1: protein molecular weight marker; lane 2: purified rNTD protein. (C). Vaccination schedule and detection. Mice received three vaccinations consisting of 5 or 10 μg of rNTD protein combined with adjuvants at 4-week intervals. Sera were collected at the indicated times to analyze the humoral immune response. Six mice from each group were sacrificed 2 weeks after the last immunization. The spleens were harvested for enzyme-linked immunospot (ELISpot), intracellular cytokine staining (ICS), and cytometric bead array (CBA) assays. In parallel experiments, the remaining mice were challenged with MERS-CoV to detect the protective effect elicited by the rNTD protein. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Rabbit-serum-derived polyclonal antibody against nucleoprotein (cat: 100213-RP02; Sino Biological Inc., Beijing, CHN) was incubated with the sections at 1:1000 dilution; goat anti-rabbit (cat: pv-9001; ZSGB-Bio, Beijing, CHN) secondary antibody was used at 1:2000, and sections were evaluated using light microscopy.

Techniques: Recombinant, Expressing, Purification, SDS Page, Western Blot, Staining, Labeling, Molecular Weight, Marker, Enzyme-linked Immunospot

Journal: Vaccine

Article Title: The recombinant N-terminal domain of spike proteins is a potential vaccine against Middle East respiratory syndrome coronavirus (MERS-CoV) infection

doi: 10.1016/j.vaccine.2016.11.064

Figure Lengend Snippet: IHC detection of virus antigen expression in mouse tissue after challenge with MERS-CoV. Lung (A–C) and trachea (D–F) sections were assessed using rabbit polyclonal antibody to MERS-CoV nucleoprotein (NP) 3 days after the MERS-CoV challenge. The dark purple spot marked the inflammatory cell infiltration, and the brown particle marked the antigen of MERS-CoV. The MERS-CoV was located mainly in the trachea. Additionally, the lung tissue showed MERS-CoV expression in all immunized groups.

Article Snippet: Rabbit-serum-derived polyclonal antibody against nucleoprotein (cat: 100213-RP02; Sino Biological Inc., Beijing, CHN) was incubated with the sections at 1:1000 dilution; goat anti-rabbit (cat: pv-9001; ZSGB-Bio, Beijing, CHN) secondary antibody was used at 1:2000, and sections were evaluated using light microscopy.

Techniques: Expressing