Journal: iScience

Article Title: BA.1 breakthrough infection elicits distinct antibody and memory B cell responses in vaccinated-only versus hybrid immunity individuals

doi: 10.1016/j.isci.2025.111962

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Article Snippet: Recombinant SARS-CoV-2 RBD (HEK)-Biotin 50ug , Miltenyi , 130-127-457.

Techniques: Recombinant, Bioassay, Luciferase, Software, Imaging, Staining, Blocking Assay

Journal: Advanced Biology

Article Title: Sars‐Cov‐2 Spike Protein‐Induced Damage of hiPSC‐Derived Cardiomyocytes

doi: 10.1002/adbi.202101327

Figure Lengend Snippet: Beating arrest and nuclear distortion of hiPSC‐CMs. A) Comparison of the rhythmic activity of hiPSC‐CMs incubated for 54 h, with or without spike protein and Ang II. Spike protein (1 µg mL ‐1 ) was added at −3 h, 100 ng mL ‐1 Ang II was added after another 3 h. Data were obtained from five positions under each condition. The blue and red significant symbols indicate the difference of the group “Ang II only/No treatment” and the group “Spike+Ang II/No treatment,” respectively. B) Immunofluorescent images of hiPSC‐CMs, showing expression of ACE2 (Green), Actin (Red) and Nucleus (Gray or Blue) under different conditions. Yellow full arrows point to cells with nuclear fragmentation while yellow hollow arrows point to cells with chromatin margining. Scale bar: 5 µm. C) The bar graph shows the mean intensity of ACE2 fluorescence from cells of 8 different positions. D) Schematic figures to show different nuclear morphology and data statistical analyses, showing quantitatively the changes in chromatin distribution. The nuclei of 120–150 cells from ten positions per condition were analyzed.

Article Snippet: The recombinant biotinylated Sars‐Cov‐2 spike protein (bs‐46003P‐Biotin) and Ang II (A9525) were purchased from Bioss and Sigma.

Techniques: Activity Assay, Incubation, Expressing, Fluorescence

Journal: Advanced Biology

Article Title: Sars‐Cov‐2 Spike Protein‐Induced Damage of hiPSC‐Derived Cardiomyocytes

doi: 10.1002/adbi.202101327

Figure Lengend Snippet: Degradation of the cellular structures of hiPSC‐CMs. A) Immunofluorescent images of hiPSC‐CMs, showing the expression of TnT (Green), Actinin (Red) under different conditions. Scale bar: 5 µm. B,C) Bar graphs at left show the mean intensity of TnT (B) and Actinin (C) fluorescence, bar graph at right shows the distribution of TnT (B) and Actinin (C) fluorescence spot areas with no treatment, spike protein or spike protein, and supplementary Ang II. D) Immunostaining images of connexin 43 (Green) and Actin (Red). Scale bar: 5 µm. E) Mean intensity of the connexin 43 fluorescence. All the data of α‐actinin, troponin T, or connexin 43 were from five different positions per condition.

Article Snippet: The recombinant biotinylated Sars‐Cov‐2 spike protein (bs‐46003P‐Biotin) and Ang II (A9525) were purchased from Bioss and Sigma.

Techniques: Expressing, Fluorescence, Immunostaining

Journal: Advanced Biology

Article Title: Sars‐Cov‐2 Spike Protein‐Induced Damage of hiPSC‐Derived Cardiomyocytes

doi: 10.1002/adbi.202101327

Figure Lengend Snippet: Schematic summaries of the effects of spike protein on hiPSC‐CMs. A) Sarcomere structures with and without spike protein treatment. Without spike protein, myosin slides along actin to contract the hiPSC‐CMs. With spike protein, the distribution of α‐actinin proteins might be disturbed and the overlap between myosin and actin became shortened. B) Damage effects of the spike protein on cells. Without spike protein, the cell‐cell contact with the help of gap junction proteins (connexin 43) enables coordinated contraction of the cardiac tissue. With spike protein, the gap junction disappeared due to mislocalization of connexin 43. When incubated with supplementary Ang II, the spike protein‐induced changes became fatal in terms of ACE2 downregulation, chromatin margination, connexin 43 expression, and sarcomere disruption, leading to cell death.

Article Snippet: The recombinant biotinylated Sars‐Cov‐2 spike protein (bs‐46003P‐Biotin) and Ang II (A9525) were purchased from Bioss and Sigma.

Techniques: Incubation, Expressing

Journal: Advanced Biology

Article Title: Sars‐Cov‐2 Spike Protein‐Induced Damage of hiPSC‐Derived Cardiomyocytes

doi: 10.1002/adbi.202101327

Figure Lengend Snippet: Overview of possible ends of shedding spike protein and evaluation process. A) After endocytosis of Sars‐Cov‐2, the spike‐ACE2 complex may: 1) Enter into the cytoplasm; 2) Totally or partially shed from the cell membrane; 3) Stay on the cell membrane. B) Workflow of cardiac patch fabrication and spike protein testing. hiPSCs are first differentiated to cardiomyocytes on a culture patch made of arrayed nanofiber membrane. They are then incubated with synthetic spike protein with or without supplementary Ang II. The beating behavior, cellular structure, and nuclear morphology of the cells are analyzed to evaluate the effect of the spike protein of Sars‐Cov‐2.

Article Snippet: The recombinant biotinylated Sars‐Cov‐2 spike protein (bs‐46003P‐Biotin) and Ang II (A9525) were purchased from Bioss and Sigma.

Techniques: Incubation

Journal: medRxiv

Article Title: Detection of SARS-CoV-2 neutralizing antibodies with a cell-free PCR assay

doi: 10.1101/2020.05.28.20105692

Figure Lengend Snippet: (A) Viral entry of SARS-CoV-2 is mediated by the binding of the spike protein to the human receptor angiotensin-converting enzyme 2 (ACE2). Disruption of this interaction forms the basis of neutralizing antibodies (NAbs). (B) Neutralization PCR assay reconstructs this interaction using a pair of S1 subunits of the spike protein- and ACE2-DNA conjugates. In the absence of NAbs, S1 and ACE2 engage with strong affinity, thereby positioning the two DNA barcodes in close proximity for subsequent ligation and PCR-amplification. On the other hand, binding of NAbs blocks S1 from ACE2, leaving the two DNA barcodes separated. Since each barcode has only one PCR primer binding site, they cannot be amplified. Therefore, the quantities of NAbs are correlated with the decrease of PCR amplicon formation.

Article Snippet: The SARS-CoV-2 spike protein (S1) containing amino acids 1–674 with an Fc-tag at the C-terminus (#31806) expressed in HEK293 cells was purchased from the Native Antigen Company (Oxford, United Kingdom).

Techniques: Binding Assay, Disruption, Neutralization, Ligation, Amplification

Journal: bioRxiv

Article Title: An alpaca nanobody neutralizes SARS-CoV-2 by blocking receptor interaction

doi: 10.1101/2020.06.02.130161

Figure Lengend Snippet: a , VSV G or SARS-CoV-2 spike pseudotyped lentivirus (PSV) was incubated with a dilution series of Ty1 or control nanobody (influenza NP-VHH1 ). Infectivity relative to cells infected with pseudotyped virus in the absence of nanobody is shown. Neutralization by Ty1 was repeated in duplicate across 6 assays, and the error bars represent the standard deviation. b , Cells were transfected with a plasmid harboring the SARS-CoV-2 spike for 24 h. Cells were fixed, permeabilized and stained with Ty1-AS635P (black and red) or left unstained (grey). Cells were analyzed by flow cytometry. Cell counts are presented as % of max (representative histogram). c , Vero E6 cells were infected with SARS-CoV-2 at a MOI of 1 for 24 hours. Cells were fixed, permeabilized and stained for DNA (blue), dsRNA (green) and with Ty1-AS635P (red). Pictures were taken by fluorescence microscopy and representative examples are shown. d , ACE2 expressing HEK293T cells were trypsinized, fixed and stained with RBD-AS635P alone, or preincubated with IAV NP-VHH1 or Ty1. Cells were analyzed by flow cytometry.

Article Snippet: SARS-CoV-2 S1-sheep-Fc (Native Antigen Company, SKU: REC31806) was used for the first two immunization, and SARS-CoV-2 RBD produced in FreeStyle 293F cells was used for the last two immunizations.

Techniques: Incubation, Infection, Virus, Neutralization, Standard Deviation, Transfection, Plasmid Preparation, Staining, Flow Cytometry, Fluorescence, Microscopy, Expressing

Journal: bioRxiv

Article Title: An alpaca nanobody neutralizes SARS-CoV-2 by blocking receptor interaction

doi: 10.1101/2020.06.02.130161

Figure Lengend Snippet: SARS-CoV-2 spike pseudotyped lentivirus (PSV) was incubated with a dilution series of Ty1 or Ty1-Fc. Infectivity relative to cells infected with pseudotyped virus in the absence of nanobody is shown. Neutralization was repeated in duplicate across 3 assays, and the error bars represent the standard deviation

Article Snippet: SARS-CoV-2 S1-sheep-Fc (Native Antigen Company, SKU: REC31806) was used for the first two immunization, and SARS-CoV-2 RBD produced in FreeStyle 293F cells was used for the last two immunizations.

Techniques: Incubation, Infection, Virus, Neutralization, Standard Deviation