Journal: Biosensors & Bioelectronics

Article Title: Magnetic beads combined with carbon black-based screen-printed electrodes for COVID-19: A reliable and miniaturized electrochemical immunosensor for SARS-CoV-2 detection in saliva

doi: 10.1016/j.bios.2020.112686

Figure Lengend Snippet: The MBs-based assay for SARS-CoV-2 detection in untreated saliva.

Article Snippet: The use of the high sensitive antibodies is one of the main task to develop high sensitive and selective analytical device, thus for the selection of antibodies, spectrophotometric ELISA was carried out to assess the reactivity of MAb and two different PAb (Sinobiological, Germany and ProSci, USA) towards two different S proteins namely SARS Coronavirus 2019 Spike Recombinant protein (1000–1200 aa) and Recombinant Spike protein SARS-CoV Spike protein, S1 subunit.

Techniques:

Journal: Biosensors & Bioelectronics

Article Title: Magnetic beads combined with carbon black-based screen-printed electrodes for COVID-19: A reliable and miniaturized electrochemical immunosensor for SARS-CoV-2 detection in saliva

doi: 10.1016/j.bios.2020.112686

Figure Lengend Snippet: A) ELISA response for two different PAb anti-SARS-CoV-2 1 μg/mL (Sinobiological and ProSci) towards two different Spike proteins coated at 2 ng/mL. B) Binding curve of colorimetric ELISA for MAb anti-SARS-CoV-2 ranging from 0.12 – 2 μg/mL. Coating of Spike protein: 2 ng/mL. C) Electrochemical response using the MBs-based assay using CB-based modified electrode (blue line) and bare electrode (black line). The mean value (n = 3) with the corresponding standard deviation was reported for each measurement. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The use of the high sensitive antibodies is one of the main task to develop high sensitive and selective analytical device, thus for the selection of antibodies, spectrophotometric ELISA was carried out to assess the reactivity of MAb and two different PAb (Sinobiological, Germany and ProSci, USA) towards two different S proteins namely SARS Coronavirus 2019 Spike Recombinant protein (1000–1200 aa) and Recombinant Spike protein SARS-CoV Spike protein, S1 subunit.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Modification, Standard Deviation

Journal: Biosensors & Bioelectronics

Article Title: Magnetic beads combined with carbon black-based screen-printed electrodes for COVID-19: A reliable and miniaturized electrochemical immunosensor for SARS-CoV-2 detection in saliva

doi: 10.1016/j.bios.2020.112686

Figure Lengend Snippet: A) Study of the signal response for evaluating PAb concentration. 10 μL of MBs, 200 μL of PAb anti SARS-CoV-2 0.5, 1, and 2 μg/mL (in PBS) + 200 μL of PAb-AP anti rabbit IgG 0.5, 1, and 2 μg/mL (in PBS) + 300 μL of tested sample, incubation time 30 min without stirring, 3 washing steps, testing a negative control and 0.16 μg/mL of S protein. B) Study of the signal response for evaluating the effect of incubation time. 10 μL of MBs, 200 μL of PAb anti SARS-CoV-2 1 μg/mL (in PBS) + 200 μL of PAb-AP anti rabbit IgG 1 μg/mL (in PBS) + 300 μL of tested sample, incubation time (15/30 min) without stirring, 3 washing steps, testing a negative control, 0.16 and 2.5 μg/mL of S protein. C) Study of the signal response as function of mixing during the incubation step. 10 μL of MBs, 200 μL of PAb anti SARS-CoV-2 1 μg/mL (in PBS) + 200 μL of PAb-AP anti rabbit IgG 1 μg/mL (in PBS) + 300 μL of tested sample, incubation time 30 min with and without stirring, 3 washing steps, testing a negative control and 0.16 and 2.5 μg/mL of S protein. D) Study of the signal response for evaluating the effect of number of washing steps. 10 μL of MBs, 200 μL of PAb anti SARS-CoV-2 1 μg/mL (in PBS) + 200 μL of PAb-AP anti rabbit IgG 1 μg/mL (in PBS) + 300 μL of tested sample, incubation time 30 min without stirring, 1/2/3 washing steps, testing a negative control and 0.16 μg/mL of S protein. The mean value (n = 3) with the corresponding standard deviation was reported for each measurement.

Article Snippet: The use of the high sensitive antibodies is one of the main task to develop high sensitive and selective analytical device, thus for the selection of antibodies, spectrophotometric ELISA was carried out to assess the reactivity of MAb and two different PAb (Sinobiological, Germany and ProSci, USA) towards two different S proteins namely SARS Coronavirus 2019 Spike Recombinant protein (1000–1200 aa) and Recombinant Spike protein SARS-CoV Spike protein, S1 subunit.

Techniques: Concentration Assay, Incubation, Negative Control, Standard Deviation

Journal: Biosensors & Bioelectronics

Article Title: Magnetic beads combined with carbon black-based screen-printed electrodes for COVID-19: A reliable and miniaturized electrochemical immunosensor for SARS-CoV-2 detection in saliva

doi: 10.1016/j.bios.2020.112686

Figure Lengend Snippet: A) Electrochemical calibration curve (inset the voltammograms) using the optimized parameters for S protein detection in buffer (black line) and in untreated saliva (red line) (n = 3). B) Electrochemical calibration curve (inset the voltammograms) for N protein detection in buffer (black line) and untreated saliva (red line) (n = 3). C) Voltammograms obtained without (black line) and with (blue line) cultured SARS-CoV-2 virus at concentration 6.5 PFU/mL using MBs-based immunoassay for S protein. D) Voltammograms obtained without (black line) and with (blue line) cultured SARS-CoV-2 virus at concentration 6.5 x 10 4 PFU/mL using MBs-based immunoassay for N protein. E) Exclusivity test on seasonal influenza virus A (H1N1) (10 2.9 TCID50 mL −1 ) and Influenza 2009 pH1N1 virus (10 4.15 TCID50 mL −1 ) in comparison to SARS-CoV-2 (6.5 PFU/mL) using assay for S protein (n = 3). F) Exclusivity test on seasonal influenza virus A (H1N1) (10 2.9 TCID50 mL −1 ) and 2009 influenza virus pH1N1 (10 4.15 TCID50 mL −1 ) in comparison to SARS-CoV-2 (6.5 x 10 4 PFU/mL) using assay for N protein (n = 3). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The use of the high sensitive antibodies is one of the main task to develop high sensitive and selective analytical device, thus for the selection of antibodies, spectrophotometric ELISA was carried out to assess the reactivity of MAb and two different PAb (Sinobiological, Germany and ProSci, USA) towards two different S proteins namely SARS Coronavirus 2019 Spike Recombinant protein (1000–1200 aa) and Recombinant Spike protein SARS-CoV Spike protein, S1 subunit.

Techniques: Cell Culture, Concentration Assay

Journal: Biosensors & Bioelectronics

Article Title: Magnetic beads combined with carbon black-based screen-printed electrodes for COVID-19: A reliable and miniaturized electrochemical immunosensor for SARS-CoV-2 detection in saliva

doi: 10.1016/j.bios.2020.112686

Figure Lengend Snippet: Overview of biosensors for SARS-CoV-2 detection.

Article Snippet: The use of the high sensitive antibodies is one of the main task to develop high sensitive and selective analytical device, thus for the selection of antibodies, spectrophotometric ELISA was carried out to assess the reactivity of MAb and two different PAb (Sinobiological, Germany and ProSci, USA) towards two different S proteins namely SARS Coronavirus 2019 Spike Recombinant protein (1000–1200 aa) and Recombinant Spike protein SARS-CoV Spike protein, S1 subunit.

Techniques: CRISPR, Amplification, Reporter Assay, SPR Assay, Transduction, Fluorescence, Immunochromatographic Assay

Journal: Cell reports

Article Title: Differences in syncytia formation by SARS-CoV-2 variants modify host chromatin accessibility and cellular senescence via TP53

doi: 10.1016/j.celrep.2023.113478

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: To generate viral stocks, Huh7.5, VeroE6, or VeroE6-ACE2-TMPRSS2 cells were inoculated with HKU5-SARS-CoV-1-S (BEI Resources #NR-48814), SARS-CoV-2 isolate USA-WA1/2020 (BEI Resources #NR-52281), MERS-CoV (BEI Resources #NR-48813), SARS-CoV-2 B.1.1.7 (alpha) isolate (BEI Resources #NR-54000), SARS-CoV-2 B.1.617.2 (delta) isolate (BEI Resources #NR-55611), SARS-CoV-2 P.1 (gamma) isolate (BEI Resources #NR-54982), or a SARS-CoV-2 BA.1 isolated from a patient at Yale New Haven Hospital at a MOI of approximately 0.01 for three days to generate a P1 stock.

Techniques: FLAG-tag, Luciferase, Virus, Recombinant, Reverse Transcription, SYBR Green Assay, Activity Assay, Software, CRISPR

Journal: bioRxiv

Article Title: Development of a Synthetic Poxvirus-Based SARS-CoV-2 Vaccine

doi: 10.1101/2020.07.01.183236

Figure Lengend Snippet: A) Schematic representation of vector construction. S and N antigen sequences (red spheres and green triangles) were inserted into sMVA fragments F2 and F3 by bacterial recombination methods in E. coli . The modified sMVA fragments of F1 and F2 with inserted antigen sequences and the unmodified sMVA fragment F1 were isolated from E. coli and co-transfected into FPV-infected BHK cells to initiate virus reconstitution. B) Schematics of single (sMVA-S, sMVA-N) and double (sMVA-N/S, sMVA-S/N) recombinant sMVA-CoV2 vectors with S and N antigen sequences inserted into commonly used MVA insertion sites (Del2, IGR69/70, Del3). All antigens were expressed via the Vaccinia mH5 promoter. C) Western Blot. BHK cells infected with the single and double recombinant sMVA-CoV2 vectors derived with FPV HP1.441 (sMVA-S/N hp, sMVA-N/S hp) or TROVAC (sMVA-S/N tv, sMVA-N/S tv, sMVA-S tv, sMVA-N tv) were evaluated for antigen expression by Western Blot using anti-S1 and N antibodies (αS1 and αN Ab). Vaccinia B5R protein was verified as infection control. Higher and lower molecular weight bands may represent mature and immature protein species. D) Flow cytometry staining. HeLa cells infected with the vaccine vectors were evaluated by cell surface and intracellular flow staining using anti-S1, S2, and N antibodies (αS1, αS2, and αN Ab). Live cells were used to evaluate cell surface antigen expression. Fixed and permeabilized cells were used to evaluate intracellular antigen expression. Anti-Vaccinia virus antibody (αVAC) was used as staining control to verify MVA protein expression. Cells infected with sMVA or wtMVA or uninfected cells were used as controls for experiments in C and D as indicated.

Article Snippet: Anti-SARS-CoV-1 S1 mouse (40150-R007, Sino Biological) and S2 rabbit (GTX632604, GeneTex) monoclonal antibodies, anti-SARS-CoV-1 N rabbit monoclonal antibody (40143-R001, Sino Biological), and anti-vaccinia rabbit polyclonal antibody (9503-2057, Bio Rad) were used in dilution 1:2,000.

Techniques: Plasmid Preparation, Modification, Isolation, Transfection, Infection, Virus, Recombinant, Western Blot, Derivative Assay, Expressing, Control, Molecular Weight, Flow Cytometry, Staining