sars cov 2 s1 s2  (Sino Biological)


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    Name:
    SARS CoV 2 2019 nCoV Spike Gene Lentiviral ORF cDNA clone expression plasmid C GFPSpark tag Codon Optimized COVID 19 Spike S1 S2 ECD Research
    Description:
    Full length Clone DNA of SARS CoV 2 2019 nCoV Spike
    Catalog Number:
    vg40589-acgln
    Product Aliases:
    coronavirus spike cDNA ORF Clone 2019-nCoV, cov spike cDNA ORF Clone 2019-nCoV, ncov RBD cDNA ORF Clone 2019-nCoV, ncov s1 cDNA ORF Clone 2019-nCoV, ncov s2 cDNA ORF Clone 2019-nCoV, ncov spike cDNA ORF Clone 2019-nCoV, NCP-CoV RBD cDNA ORF Clone 2019-nCoV, NCP-CoV s1 cDNA ORF Clone 2019-nCoV, NCP-CoV s2 cDNA ORF Clone 2019-nCoV, NCP-CoV Spike cDNA ORF Clone 2019-nCoV, novel coronavirus RBD cDNA ORF Clone 2019-nCoV, novel coronavirus s1 cDNA ORF Clone 2019-nCoV, novel coronavirus s2 cDNA ORF Clone 2019-nCoV, novel coronavirus spike cDNA ORF Clone 2019-nCoV, RBD cDNA ORF Clone 2019-nCoV, S1 cDNA ORF Clone 2019-nCoV, S2 cDNA ORF Clone 2019-nCoV, Spike RBD cDNA ORF Clone 2019-nCoV
    Price:
    645.0
    Applications:
    Stable or Transient mammalian expression
    Size:
    1Unit
    Category:
    cDNA Clone
    Molecule Name:
    Spike
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    Structured Review

    Sino Biological sars cov 2 s1 s2
    SARS CoV 2 2019 nCoV Spike Gene Lentiviral ORF cDNA clone expression plasmid C GFPSpark tag Codon Optimized COVID 19 Spike S1 S2 ECD Research
    Full length Clone DNA of SARS CoV 2 2019 nCoV Spike
    https://www.bioz.com/result/sars cov 2 s1 s2/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 s1 s2 - by Bioz Stars, 2021-02
    94/100 stars

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    Related Articles

    Recombinant:

    Article Title: Evaluating SARS-CoV-2 spike and nucleocapsid proteins as targets for antibody detection in severe and mild COVID-19 cases using a Luminex bead-based assay
    Article Snippet: .. 2.3 Luminex bead-based IgG immunoassayWe ordered commercially available recombinant NP, RBD, S1 and the complete Spike unit (S1S2) antigens derived from SARS-CoV-2 at Sino Biological (BIOCONNECT, Huissen, The Netherlands). .. Each antigen was coupled to a maximum of 1.25 × 10 ( ) paramagnetic MAGPLEX COOH-microsphere beads from Luminex Corporation (Austin, TX), as described previously ( ; ).

    Article Title: A therapeutic neutralizing antibody targeting receptor binding domain of SARS-CoV-2 spike protein
    Article Snippet: .. Recombinant proteins for RBD and its mutants (A435S, F342L, G476S, K458R, N354D, V367F, V483A, W436R), SARS-CoV S1, HCoV-HKU1 S1, and MERS-CoV RBD were commercial products (Sino Biological). .. mAb-neutralizing assays To evaluate the neutralizing activity of monoclonal antibodies, plaque reduction neutralizing tests for SARS-CoV-2 were performed as described previously .

    Article Title: Therapeutic activity of an inhaled potent SARS-CoV-2 neutralizing human monoclonal antibody in hamsters
    Article Snippet: .. Recombinant proteins used include SARS-CoV-2 S1+S2 (Sino Biological, Wayne, PA), SARS-CoV-2 D614G S1 (Sino Biological), SARS-CoV-1 S (BEI Resources), SARS-CoV-2 Nucleocapsid (Sino Biological), and HepG2 whole cell lysate (Abcam, Cambridge, MA). .. Purified hmAbs were diluted in PBS, and binding was detected with HRP-conjugated anti-human IgG (Jackson ImmunoResearch, West Grove, PA).

    Article Title: BANCOVID, the first D614G variant mRNA-based vaccine candidate against SARS-CoV-2 elicits neutralizing antibody and balanced cellular immune response
    Article Snippet: .. Peptide pool preparationDissolve 40 µg of SARS-CoV-2 Spike S1+S2 ECD His recombinant protein (Sino Biological, China), S2 ECD-His Recombinant Protein (Sino Biological, China), and RBD-His Recombinant Protein (Sino Biological, China) in 50mM ammonium bicarbonate (Wako Pure Chemicals Industries Ltd., Japan), pH 8 containing 8M urea (ThermoFisher Scientific, USA). .. After dissolving, add 500 mM DTT (ThermoFisher Scientific, USA) to the solution to a final concentration of 20 mM (1:25 dilution) and mix briefly; incubate at 60 °C for 1 hour.

    Binding Assay:

    Article Title: Methylene Blue Inhibits the SARS-CoV-2 Spike–ACE2 Protein-Protein Interaction–a Mechanism that can Contribute to its Antiviral Activity Against COVID-19
    Article Snippet: .. ACE2-Fc and SARS-CoV-2 S1 or RBD with His tag proteins used in the binding assays were obtained from Sino Biological (Wayne, PA, United States); catalog no. 10108-H05H, 40591-V08H, and 40592-V08H). .. Binding inhibition assays were performed in a 96-well cell-free format similar to the one described before ( ; ; ; ).

    Derivative Assay:

    Article Title: Evaluating SARS-CoV-2 spike and nucleocapsid proteins as targets for antibody detection in severe and mild COVID-19 cases using a Luminex bead-based assay
    Article Snippet: .. 2.3 Luminex bead-based IgG immunoassayWe ordered commercially available recombinant NP, RBD, S1 and the complete Spike unit (S1S2) antigens derived from SARS-CoV-2 at Sino Biological (BIOCONNECT, Huissen, The Netherlands). .. Each antigen was coupled to a maximum of 1.25 × 10 ( ) paramagnetic MAGPLEX COOH-microsphere beads from Luminex Corporation (Austin, TX), as described previously ( ; ).

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  • 94
    Sino Biological sars cov 2
    Structural conservation of SARS-CoV RBD. RBD is shown as colored surface. ACE2 is shown as gray cartoon. The three surface mutation sites (i.e. N354D, D364Y, and V367F) observed in <t>SARS-CoV-2</t> RBD are labeled. Mutation F342L is buried and not shown here.
    Sars Cov 2, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sars cov 2/product/Sino Biological
    Average 94 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 - by Bioz Stars, 2021-02
    94/100 stars
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    90
    Sino Biological sars cov 2 spike gene
    IgM, IgA, and IgG depletion in plasma samples from convalescent donors (A–C) Efficacy of the specific isotype depletion assessed by ELISA for total IgM, IgA, and IgG. All plasma samples were diluted 5-fold before depletion. (A) IgM concentration in non-depleted, IgM-depleted, IgA-depleted, and IgG-depleted plasma samples, measured with an anti-human IgM (μ-chain specific) as the capture antibody. (B) IgA concentration measured on the same plasma samples with an anti-human IgA (α-chain specific). (C) IgG concentration measured with anti-human an IgG (γ-chain specific). (D–G) Efficacy of <t>SARS-CoV-2-specific</t> antibody depletion as assessed by SARS-CoV-2 RBD ELISA. (D) Level of total (pan-Ig) anti-SARS-CoV-2, RBD-specific antibodies in non-depleted, IgM-depleted, IgA-depleted, and IgG-depleted plasma samples. (E) Level of IgM-specific anti-RBD. (F) Level of IgA-specific anti-RBD. (G) Level of IgG-specific anti-RBD. (H–K) Efficacy of full S glycoprotein-specific antibody depletion measured by flow cytometry. (H) Level of total (pan-Ig) anti-SARS-CoV-2 S-specific antibodies in non-depleted, IgM-depleted, IgA-depleted, and IgG-depleted plasma samples. (I) Level of IgM-specific anti-S. (J) Level of IgA-specific anti-S. (K) Level of IgG-specific anti-S. Red dashed lines represent the average signal given by negative controls taken from non-infected patients. Asterisks indicate the level of statistical significance obtained by a Dunn’s test. ∗∗∗∗ p
    Sars Cov 2 Spike Gene, supplied by Sino Biological, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sars cov 2 spike gene/product/Sino Biological
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 spike gene - by Bioz Stars, 2021-02
    90/100 stars
      Buy from Supplier

    Image Search Results


    Structural conservation of SARS-CoV RBD. RBD is shown as colored surface. ACE2 is shown as gray cartoon. The three surface mutation sites (i.e. N354D, D364Y, and V367F) observed in SARS-CoV-2 RBD are labeled. Mutation F342L is buried and not shown here.

    Journal: bioRxiv

    Article Title: SARS-CoV-2 and SARS-CoV Spike-RBD Structure and Receptor Binding Comparison and Potential Implications on Neutralizing Antibody and Vaccine Development

    doi: 10.1101/2020.02.16.951723

    Figure Lengend Snippet: Structural conservation of SARS-CoV RBD. RBD is shown as colored surface. ACE2 is shown as gray cartoon. The three surface mutation sites (i.e. N354D, D364Y, and V367F) observed in SARS-CoV-2 RBD are labeled. Mutation F342L is buried and not shown here.

    Article Snippet: Reagents, recombinant proteins and antibodies Recombinant S1 proteins of SARS-CoV-2 (Cat: 40591-V08H), SARS-CoV (Cat: 40150-V08B1) and MERS-CoV (Cat:40069-V08H), recombinant RBD protein of SARS-CoV (Cat: 40150-V31B2), transfection reagent Sinofection (Cat: STF02), mammalian expression plasmids of full length S or RBD protein of SARS-CoV-2 (Cat: VG40589-UT, Wuhan/IVDC-HB-01/2019) and SARS-CoV (Cat: VG40150-G-N, CUHK-W1), ACE2 (Cat: HG10108-UT), polyclonal antibodies against SARS-CoV RP01 (Cat: 40150-RP01) and T52 (Cat: 40150-T52) were purchased from Sino Biological.

    Techniques: Mutagenesis, Labeling

    Structure similarity between SARS-CoV-2 RBD and SARS-CoV RBD. RBD is shown in a space-filled model with colored surface. ACE2 is shown as gray tube model. The three glycosylation sites in SARS-CoV are labeled. Note that N 357 ST in SARS-CoV is changed to N 370 SA in SARS-CoV-2, which is different from the NXS/T pattern required for glycosylation, and hence this site is more likely to be unglycosylated. The two possible cross-reactive regions are marked with yellow circles.

    Journal: bioRxiv

    Article Title: SARS-CoV-2 and SARS-CoV Spike-RBD Structure and Receptor Binding Comparison and Potential Implications on Neutralizing Antibody and Vaccine Development

    doi: 10.1101/2020.02.16.951723

    Figure Lengend Snippet: Structure similarity between SARS-CoV-2 RBD and SARS-CoV RBD. RBD is shown in a space-filled model with colored surface. ACE2 is shown as gray tube model. The three glycosylation sites in SARS-CoV are labeled. Note that N 357 ST in SARS-CoV is changed to N 370 SA in SARS-CoV-2, which is different from the NXS/T pattern required for glycosylation, and hence this site is more likely to be unglycosylated. The two possible cross-reactive regions are marked with yellow circles.

    Article Snippet: Reagents, recombinant proteins and antibodies Recombinant S1 proteins of SARS-CoV-2 (Cat: 40591-V08H), SARS-CoV (Cat: 40150-V08B1) and MERS-CoV (Cat:40069-V08H), recombinant RBD protein of SARS-CoV (Cat: 40150-V31B2), transfection reagent Sinofection (Cat: STF02), mammalian expression plasmids of full length S or RBD protein of SARS-CoV-2 (Cat: VG40589-UT, Wuhan/IVDC-HB-01/2019) and SARS-CoV (Cat: VG40150-G-N, CUHK-W1), ACE2 (Cat: HG10108-UT), polyclonal antibodies against SARS-CoV RP01 (Cat: 40150-RP01) and T52 (Cat: 40150-T52) were purchased from Sino Biological.

    Techniques: Labeling

    Cross-reactivity and neutralization efficiency of SARS nAbs against SARS-CoV-2. A. Binding of SARS nAbs to SARS-CoV S1 protein were tested by ELISA. Recombinant S1 protein of SARS-CoV were coated on plates, serial diluted nAbs were added for binding to recombinant S1 protein. B. Binding of SARS nAbs to SARS-CoV-2 S1 protein were tested by ELSIA. Recombinant S1 protein of SARS-CoV-2 were coated on plates, serial diluted nAbs were added for binding to recombinant S1 protein. C. Neutralization of SARS-CoV nAbs against SARS-CoV-2 PSV. D. Antibody competition with SARS-CoV RBD binding to ACE2. Recombinant SARS-CoV RBD protein was coated on plates, nAbs and recombinant ACE2 were then added for RBD binding competition measurements.

    Journal: bioRxiv

    Article Title: SARS-CoV-2 and SARS-CoV Spike-RBD Structure and Receptor Binding Comparison and Potential Implications on Neutralizing Antibody and Vaccine Development

    doi: 10.1101/2020.02.16.951723

    Figure Lengend Snippet: Cross-reactivity and neutralization efficiency of SARS nAbs against SARS-CoV-2. A. Binding of SARS nAbs to SARS-CoV S1 protein were tested by ELISA. Recombinant S1 protein of SARS-CoV were coated on plates, serial diluted nAbs were added for binding to recombinant S1 protein. B. Binding of SARS nAbs to SARS-CoV-2 S1 protein were tested by ELSIA. Recombinant S1 protein of SARS-CoV-2 were coated on plates, serial diluted nAbs were added for binding to recombinant S1 protein. C. Neutralization of SARS-CoV nAbs against SARS-CoV-2 PSV. D. Antibody competition with SARS-CoV RBD binding to ACE2. Recombinant SARS-CoV RBD protein was coated on plates, nAbs and recombinant ACE2 were then added for RBD binding competition measurements.

    Article Snippet: Reagents, recombinant proteins and antibodies Recombinant S1 proteins of SARS-CoV-2 (Cat: 40591-V08H), SARS-CoV (Cat: 40150-V08B1) and MERS-CoV (Cat:40069-V08H), recombinant RBD protein of SARS-CoV (Cat: 40150-V31B2), transfection reagent Sinofection (Cat: STF02), mammalian expression plasmids of full length S or RBD protein of SARS-CoV-2 (Cat: VG40589-UT, Wuhan/IVDC-HB-01/2019) and SARS-CoV (Cat: VG40150-G-N, CUHK-W1), ACE2 (Cat: HG10108-UT), polyclonal antibodies against SARS-CoV RP01 (Cat: 40150-RP01) and T52 (Cat: 40150-T52) were purchased from Sino Biological.

    Techniques: Neutralization, Binding Assay, Enzyme-linked Immunosorbent Assay, Recombinant

    Sequence analysis and structure modeling of SARS-CoV-2 RBD and SARS-CoV RBD and their interactions with ACE2. A. RBD sequence alignment of SARS-CoV and SARS-CoV-2, highlighting the predominant residues that contribute to the interactions with ACE2. The distinct interactions of RBD and ACE2 for the two viruses are indicated by the down-pointing orange triangles and up-pointing red triangles, respectively. RBM residues are underlined. The one-residue insertion is indicated by the red arrow. Asterisks indicate positions of fully conserved residues. Colons indicate positions of strictly conserved residues. Periods indicate positions of weakly conserved residues. B. Conformational comparison between the RBD-ACE2 complex structures for SARS-CoV-2 and SARS-CoV. The RBD and ACE2 structures in the SARS-CoV-2 RBD-ACE2 complex model are shown as orange and pink tubes, respectively. The RBD and ACE2 structures in the optimized SARS-CoV RBD-ACE2 complex structure are shown as blue and green tubes, respectively. The location of noticeable subtle conformational difference is indicated by an arrow. C. Distinct interaction patterns in the SARS-CoV-2 and SARS-CoV RBD-ACE2 interfaces. Structures of RBD and ACE2 are shown as cartoon in pink and green colors, respectively. The side chains of the residues in both protein components, representing their unique interactions, are shown as sticks. Polar interactions (salt-bridge and hydrogen bond) are shown as blue dash line. Non-polar interactions (π-stack, π-anion, and hydrophobic interactions) are shown as orange dash line.

    Journal: bioRxiv

    Article Title: SARS-CoV-2 and SARS-CoV Spike-RBD Structure and Receptor Binding Comparison and Potential Implications on Neutralizing Antibody and Vaccine Development

    doi: 10.1101/2020.02.16.951723

    Figure Lengend Snippet: Sequence analysis and structure modeling of SARS-CoV-2 RBD and SARS-CoV RBD and their interactions with ACE2. A. RBD sequence alignment of SARS-CoV and SARS-CoV-2, highlighting the predominant residues that contribute to the interactions with ACE2. The distinct interactions of RBD and ACE2 for the two viruses are indicated by the down-pointing orange triangles and up-pointing red triangles, respectively. RBM residues are underlined. The one-residue insertion is indicated by the red arrow. Asterisks indicate positions of fully conserved residues. Colons indicate positions of strictly conserved residues. Periods indicate positions of weakly conserved residues. B. Conformational comparison between the RBD-ACE2 complex structures for SARS-CoV-2 and SARS-CoV. The RBD and ACE2 structures in the SARS-CoV-2 RBD-ACE2 complex model are shown as orange and pink tubes, respectively. The RBD and ACE2 structures in the optimized SARS-CoV RBD-ACE2 complex structure are shown as blue and green tubes, respectively. The location of noticeable subtle conformational difference is indicated by an arrow. C. Distinct interaction patterns in the SARS-CoV-2 and SARS-CoV RBD-ACE2 interfaces. Structures of RBD and ACE2 are shown as cartoon in pink and green colors, respectively. The side chains of the residues in both protein components, representing their unique interactions, are shown as sticks. Polar interactions (salt-bridge and hydrogen bond) are shown as blue dash line. Non-polar interactions (π-stack, π-anion, and hydrophobic interactions) are shown as orange dash line.

    Article Snippet: Reagents, recombinant proteins and antibodies Recombinant S1 proteins of SARS-CoV-2 (Cat: 40591-V08H), SARS-CoV (Cat: 40150-V08B1) and MERS-CoV (Cat:40069-V08H), recombinant RBD protein of SARS-CoV (Cat: 40150-V31B2), transfection reagent Sinofection (Cat: STF02), mammalian expression plasmids of full length S or RBD protein of SARS-CoV-2 (Cat: VG40589-UT, Wuhan/IVDC-HB-01/2019) and SARS-CoV (Cat: VG40150-G-N, CUHK-W1), ACE2 (Cat: HG10108-UT), polyclonal antibodies against SARS-CoV RP01 (Cat: 40150-RP01) and T52 (Cat: 40150-T52) were purchased from Sino Biological.

    Techniques: Sequencing

    Measurements of SARS-CoV-2 and SARS-CoV S1 binding to ACE2. A. Serial diluted recombinant S1 proteins of SARS-CoV-2, SARS-CoV and MERS-CoV were coated on 96 well plates, incubated with the recombinant Fc-tagged ACE2 (ACE2-Fc) for binding evaluation. B. Recombinant S1 proteins of SARS-CoV-2 and SARS-CoV were incubated with 293T-ACE2 cells and subjected to FACS evaluation for binding.

    Journal: bioRxiv

    Article Title: SARS-CoV-2 and SARS-CoV Spike-RBD Structure and Receptor Binding Comparison and Potential Implications on Neutralizing Antibody and Vaccine Development

    doi: 10.1101/2020.02.16.951723

    Figure Lengend Snippet: Measurements of SARS-CoV-2 and SARS-CoV S1 binding to ACE2. A. Serial diluted recombinant S1 proteins of SARS-CoV-2, SARS-CoV and MERS-CoV were coated on 96 well plates, incubated with the recombinant Fc-tagged ACE2 (ACE2-Fc) for binding evaluation. B. Recombinant S1 proteins of SARS-CoV-2 and SARS-CoV were incubated with 293T-ACE2 cells and subjected to FACS evaluation for binding.

    Article Snippet: Reagents, recombinant proteins and antibodies Recombinant S1 proteins of SARS-CoV-2 (Cat: 40591-V08H), SARS-CoV (Cat: 40150-V08B1) and MERS-CoV (Cat:40069-V08H), recombinant RBD protein of SARS-CoV (Cat: 40150-V31B2), transfection reagent Sinofection (Cat: STF02), mammalian expression plasmids of full length S or RBD protein of SARS-CoV-2 (Cat: VG40589-UT, Wuhan/IVDC-HB-01/2019) and SARS-CoV (Cat: VG40150-G-N, CUHK-W1), ACE2 (Cat: HG10108-UT), polyclonal antibodies against SARS-CoV RP01 (Cat: 40150-RP01) and T52 (Cat: 40150-T52) were purchased from Sino Biological.

    Techniques: Binding Assay, Recombinant, Incubation, FACS

    Conformational Effects of Trypsin Treatment of Spikes Follow the hACE2-Dependent Activation Pathway (A–C) The serine protease trypsin remodels conformational landscape of spike proteins toward down-stream conformations on the path of hACE2-dependent activation. (A and B) The FRET histogram (A) and TDP (B) of spike proteins on HIV-1 lentivirus particles in the presence of 50 μg/mL trypsin. (C) An experiment as in (A), for spikes in the presence of both 50 μg/mL trypsin and 200 μg/mL hACE2. (D) Three-dimensional presentations of FRET histograms of spike proteins on the virus in the presence and the absence of hACE2 and trypsin. FRET histograms represent mean ± SEM, determined from three randomly assigned populations of FRET traces. For evaluated state occupancies, see Table S1 . (E and F) Trypsin enhances SARS-CoV-2 spike-mediated hACE2-dependent virus-cell fusion. (E) Assay design to monitor virus-cell fusion using the HiBit and LgBiT split NanoLuc system ( Yamamoto et al., 2019 ). Vpr-HiBit was packaged into lentiviral particles carrying SARS-CoV-2 spike (LV_Spike). HEK293 target cells transiently expressing LgBiT tagged to PH domain of human phospholipase Cδ at the N terminus alone or together with hACE2. hACE2-dependent virus-cell fusion was determined by monitoring reconstituted NanoLuc activity in target cells 24 h after infection. (F) Normalized relative luciferase units (RLU; mean ± SD, two replicates with quadruplicates) measured 24 h post-infection to quantify virus-cell fusion in stated target cells after treating viruses with or with indicated amounts of trypsin for 15–20 min at 37°C. NanoLuc activities were normalized to luciferase activity detected in uninfected target cells. p values derived from unpaired t test; ∗∗∗∗ corresponds to p

    Journal: Cell Host & Microbe

    Article Title: Real-Time Conformational Dynamics of SARS-CoV-2 Spikes on Virus Particles

    doi: 10.1016/j.chom.2020.11.001

    Figure Lengend Snippet: Conformational Effects of Trypsin Treatment of Spikes Follow the hACE2-Dependent Activation Pathway (A–C) The serine protease trypsin remodels conformational landscape of spike proteins toward down-stream conformations on the path of hACE2-dependent activation. (A and B) The FRET histogram (A) and TDP (B) of spike proteins on HIV-1 lentivirus particles in the presence of 50 μg/mL trypsin. (C) An experiment as in (A), for spikes in the presence of both 50 μg/mL trypsin and 200 μg/mL hACE2. (D) Three-dimensional presentations of FRET histograms of spike proteins on the virus in the presence and the absence of hACE2 and trypsin. FRET histograms represent mean ± SEM, determined from three randomly assigned populations of FRET traces. For evaluated state occupancies, see Table S1 . (E and F) Trypsin enhances SARS-CoV-2 spike-mediated hACE2-dependent virus-cell fusion. (E) Assay design to monitor virus-cell fusion using the HiBit and LgBiT split NanoLuc system ( Yamamoto et al., 2019 ). Vpr-HiBit was packaged into lentiviral particles carrying SARS-CoV-2 spike (LV_Spike). HEK293 target cells transiently expressing LgBiT tagged to PH domain of human phospholipase Cδ at the N terminus alone or together with hACE2. hACE2-dependent virus-cell fusion was determined by monitoring reconstituted NanoLuc activity in target cells 24 h after infection. (F) Normalized relative luciferase units (RLU; mean ± SD, two replicates with quadruplicates) measured 24 h post-infection to quantify virus-cell fusion in stated target cells after treating viruses with or with indicated amounts of trypsin for 15–20 min at 37°C. NanoLuc activities were normalized to luciferase activity detected in uninfected target cells. p values derived from unpaired t test; ∗∗∗∗ corresponds to p

    Article Snippet: Construction of Full-Length Tagged SARS-CoV-2 Spike (S)A full-length wild-type pCMV3-SARS-CoV-2 Spike (S1+S2)-long (termed as pCMV-S, codon-optimized, Sino Biological, cat # VG40589-UT) plasmid was used as a template to generate tagged pCMV-S.

    Techniques: Activation Assay, Expressing, Activity Assay, Infection, Luciferase, Derivative Assay

    IgM, IgA, and IgG depletion in plasma samples from convalescent donors (A–C) Efficacy of the specific isotype depletion assessed by ELISA for total IgM, IgA, and IgG. All plasma samples were diluted 5-fold before depletion. (A) IgM concentration in non-depleted, IgM-depleted, IgA-depleted, and IgG-depleted plasma samples, measured with an anti-human IgM (μ-chain specific) as the capture antibody. (B) IgA concentration measured on the same plasma samples with an anti-human IgA (α-chain specific). (C) IgG concentration measured with anti-human an IgG (γ-chain specific). (D–G) Efficacy of SARS-CoV-2-specific antibody depletion as assessed by SARS-CoV-2 RBD ELISA. (D) Level of total (pan-Ig) anti-SARS-CoV-2, RBD-specific antibodies in non-depleted, IgM-depleted, IgA-depleted, and IgG-depleted plasma samples. (E) Level of IgM-specific anti-RBD. (F) Level of IgA-specific anti-RBD. (G) Level of IgG-specific anti-RBD. (H–K) Efficacy of full S glycoprotein-specific antibody depletion measured by flow cytometry. (H) Level of total (pan-Ig) anti-SARS-CoV-2 S-specific antibodies in non-depleted, IgM-depleted, IgA-depleted, and IgG-depleted plasma samples. (I) Level of IgM-specific anti-S. (J) Level of IgA-specific anti-S. (K) Level of IgG-specific anti-S. Red dashed lines represent the average signal given by negative controls taken from non-infected patients. Asterisks indicate the level of statistical significance obtained by a Dunn’s test. ∗∗∗∗ p

    Journal: Cell Reports

    Article Title: Major role of IgM in the neutralizing activity of convalescent plasma against SARS-CoV-2

    doi: 10.1016/j.celrep.2021.108790

    Figure Lengend Snippet: IgM, IgA, and IgG depletion in plasma samples from convalescent donors (A–C) Efficacy of the specific isotype depletion assessed by ELISA for total IgM, IgA, and IgG. All plasma samples were diluted 5-fold before depletion. (A) IgM concentration in non-depleted, IgM-depleted, IgA-depleted, and IgG-depleted plasma samples, measured with an anti-human IgM (μ-chain specific) as the capture antibody. (B) IgA concentration measured on the same plasma samples with an anti-human IgA (α-chain specific). (C) IgG concentration measured with anti-human an IgG (γ-chain specific). (D–G) Efficacy of SARS-CoV-2-specific antibody depletion as assessed by SARS-CoV-2 RBD ELISA. (D) Level of total (pan-Ig) anti-SARS-CoV-2, RBD-specific antibodies in non-depleted, IgM-depleted, IgA-depleted, and IgG-depleted plasma samples. (E) Level of IgM-specific anti-RBD. (F) Level of IgA-specific anti-RBD. (G) Level of IgG-specific anti-RBD. (H–K) Efficacy of full S glycoprotein-specific antibody depletion measured by flow cytometry. (H) Level of total (pan-Ig) anti-SARS-CoV-2 S-specific antibodies in non-depleted, IgM-depleted, IgA-depleted, and IgG-depleted plasma samples. (I) Level of IgM-specific anti-S. (J) Level of IgA-specific anti-S. (K) Level of IgG-specific anti-S. Red dashed lines represent the average signal given by negative controls taken from non-infected patients. Asterisks indicate the level of statistical significance obtained by a Dunn’s test. ∗∗∗∗ p

    Article Snippet: For the generation of 293T cells stably expressing SARS-CoV-2 Spike the same technique than previously described has been used ( ): VSV-G pseudotyped lentivirus packaging the SARS-CoV-2 Spike gene was produced in 293T using a third-generation lentiviral vector system.

    Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Flow Cytometry, Infection

    Role of IgM, IgA, and IgG in neutralization (A) Comparison of the SARS-CoV-2 pseudoviral inhibitory dilution (ID 50 ) of all plasma samples. (B–D) ID 50 of plasma from each convalescent donor before and after IgM (B), IgA (C), and IgG (D) depletion. (E) Fold decrease (isotype-depleted versus non-depleted plasma) in ID 50 measured by SARS-CoV-2 pseudoviral particle neutralization. (F and G) Microneutralization assay with infectious wild-type SARS-CoV-2 performed on non-depleted and isotype-depleted plasma from 10 donors. Mean percentage of infection (F) and ID 50 observed from plasma from the 10 donors (G). (H) ID 50 obtained with the pseudoviral particle neutralization assay for the samples in (F)–(G). Asterisks indicate the level of statistical significance obtained by a Wilcoxon signed rank test. n.s., not significant. ∗ p

    Journal: Cell Reports

    Article Title: Major role of IgM in the neutralizing activity of convalescent plasma against SARS-CoV-2

    doi: 10.1016/j.celrep.2021.108790

    Figure Lengend Snippet: Role of IgM, IgA, and IgG in neutralization (A) Comparison of the SARS-CoV-2 pseudoviral inhibitory dilution (ID 50 ) of all plasma samples. (B–D) ID 50 of plasma from each convalescent donor before and after IgM (B), IgA (C), and IgG (D) depletion. (E) Fold decrease (isotype-depleted versus non-depleted plasma) in ID 50 measured by SARS-CoV-2 pseudoviral particle neutralization. (F and G) Microneutralization assay with infectious wild-type SARS-CoV-2 performed on non-depleted and isotype-depleted plasma from 10 donors. Mean percentage of infection (F) and ID 50 observed from plasma from the 10 donors (G). (H) ID 50 obtained with the pseudoviral particle neutralization assay for the samples in (F)–(G). Asterisks indicate the level of statistical significance obtained by a Wilcoxon signed rank test. n.s., not significant. ∗ p

    Article Snippet: For the generation of 293T cells stably expressing SARS-CoV-2 Spike the same technique than previously described has been used ( ): VSV-G pseudotyped lentivirus packaging the SARS-CoV-2 Spike gene was produced in 293T using a third-generation lentiviral vector system.

    Techniques: Neutralization, Microneutralization Assay, Infection