pcaggs  (Sino Biological)


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    Name:
    SARS CoV 2 2019 nCoV Spike ORF mammalian expression plasmid Codon Optimized COVID 19 Spike Research
    Description:
    Full length Clone DNA of SARS CoV 2 2019 nCoV Spike
    Catalog Number:
    VG40589-UT
    Price:
    670.0
    Category:
    cDNA Clone
    Applications:
    Stable or Transient mammalian expression
    Size:
    1Unit
    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
    Molecule Name:
    Spike
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    Structured Review

    Sino Biological pcaggs
    SARS CoV 2 2019 nCoV Spike ORF mammalian expression plasmid Codon Optimized COVID 19 Spike Research
    Full length Clone DNA of SARS CoV 2 2019 nCoV Spike
    https://www.bioz.com/result/pcaggs/product/Sino Biological
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcaggs - by Bioz Stars, 2021-04
    98/100 stars

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    Related Articles

    Transfection:

    Article Title: An antibody-dependent enhancement (ADE) activity eliminated neutralizing antibody with potent prophylactic and therapeutic efficacy against SARS-CoV-2 in rhesus monkeys
    Article Snippet: Flow cytometry assay The binding of MW05 and MW07 to S protein expression on cell surface was assessed by FACS. .. HEK293 cells were transiently transfected by SARS-CoV-2 Spike expression plasmid (Cat: VG40589-UT, Sino Biological) for 24 to 48 hours. ..

    Article Title: SARS-CoV-2 and SARS-CoV Spike-RBD Structure and Receptor Binding Comparison and Potential Implications on Neutralizing Antibody and Vaccine Development
    Article Snippet: .. Reagents, recombinant proteins and antibodiesRecombinant 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. ..

    Article Title: Development of a multi-antigenic SARS-CoV-2 vaccine candidate using a synthetic poxvirus platform
    Article Snippet: SARS-CoV-2 pseudovirus production The day before transfection, HEK293T/17 were seeded in a 15-cm dish at a density of 5 × 106 cells in DMEM supplemented with 10% heat inactivated FBS, non-essential amino acids, HEPES, and glutamine . .. Next day, cells were transfected with a mix of packaging vector (pALDI-Lenti System, Aldevron), luciferase reporter vector, and a plasmid encoding for the wild-type SARS-CoV2 Spike protein (VG40589-UT, Sino Biological) or vesicular stomatitis virus G (VSV-G, Aldevron), using FuGENE6 (Roche) as a transfection reagent:DNA ratio of 3:1, according to manufacturer’s protocol. .. Sixteen hours post-transfection, the media was replaced and cells were incubated for an additional 24–72 h. Supernatants were harvested at 24, 48, and 72 h, clarified by centrifugation at 1500 r.p.m. for 5 min and filtered using a sterile 0.22-µm pore size filter.

    Expressing:

    Article Title: An antibody-dependent enhancement (ADE) activity eliminated neutralizing antibody with potent prophylactic and therapeutic efficacy against SARS-CoV-2 in rhesus monkeys
    Article Snippet: Flow cytometry assay The binding of MW05 and MW07 to S protein expression on cell surface was assessed by FACS. .. HEK293 cells were transiently transfected by SARS-CoV-2 Spike expression plasmid (Cat: VG40589-UT, Sino Biological) for 24 to 48 hours. ..

    Article Title: Naturally mutated spike proteins of SARS-CoV-2 variants show differential levels of cell entry
    Article Snippet: The SARS-CoV S expression plasmid pC-SARS-S was created by inserting the BsiWI/XhoI-digested PCR-amplified SARS-CoV S fragment of CMV/R-SARS-S into the corresponding site of pCAGGS. .. The SARS-CoV-2 S expression plasmid pC-SARS2-S was created by inserting the Acc65I/NotI-digested PCR-amplified SARS-CoV-2 S fragment of pCMV3-2019-nCoV-Spike(S1+S2)-long (Sino Biological; VG40589-UT) into the corresponding site of pCAGGS. .. The SARS-CoV-2 S mutants (pC-SARS2-S-H49Y, pC-SARS2-S-V367F, pC-SARS2-S-G476S, pC-SARS2-S-V483A, pC-SARS2-S-D614G, or pC-SARS2-S-C1247A), in which positions 49, 367, 476, 483, 614, or 1247 of the S protein were mutated from histidine to tyrosine, valine to phenylalanine, glycine to serine, valine to alanine, aspartic acid to glycine, or cysteine to alanine, respectively, were created by inserting overlapping PCR fragments into Acc65I/NotI-digested pCAGGS.

    Article Title: SARS-CoV-2 and SARS-CoV Spike-RBD Structure and Receptor Binding Comparison and Potential Implications on Neutralizing Antibody and Vaccine Development
    Article Snippet: .. Reagents, recombinant proteins and antibodiesRecombinant 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. ..

    Plasmid Preparation:

    Article Title: An antibody-dependent enhancement (ADE) activity eliminated neutralizing antibody with potent prophylactic and therapeutic efficacy against SARS-CoV-2 in rhesus monkeys
    Article Snippet: Flow cytometry assay The binding of MW05 and MW07 to S protein expression on cell surface was assessed by FACS. .. HEK293 cells were transiently transfected by SARS-CoV-2 Spike expression plasmid (Cat: VG40589-UT, Sino Biological) for 24 to 48 hours. ..

    Article Title: Naturally mutated spike proteins of SARS-CoV-2 variants show differential levels of cell entry
    Article Snippet: The SARS-CoV S expression plasmid pC-SARS-S was created by inserting the BsiWI/XhoI-digested PCR-amplified SARS-CoV S fragment of CMV/R-SARS-S into the corresponding site of pCAGGS. .. The SARS-CoV-2 S expression plasmid pC-SARS2-S was created by inserting the Acc65I/NotI-digested PCR-amplified SARS-CoV-2 S fragment of pCMV3-2019-nCoV-Spike(S1+S2)-long (Sino Biological; VG40589-UT) into the corresponding site of pCAGGS. .. The SARS-CoV-2 S mutants (pC-SARS2-S-H49Y, pC-SARS2-S-V367F, pC-SARS2-S-G476S, pC-SARS2-S-V483A, pC-SARS2-S-D614G, or pC-SARS2-S-C1247A), in which positions 49, 367, 476, 483, 614, or 1247 of the S protein were mutated from histidine to tyrosine, valine to phenylalanine, glycine to serine, valine to alanine, aspartic acid to glycine, or cysteine to alanine, respectively, were created by inserting overlapping PCR fragments into Acc65I/NotI-digested pCAGGS.

    Article Title: Development of a multi-antigenic SARS-CoV-2 vaccine candidate using a synthetic poxvirus platform
    Article Snippet: SARS-CoV-2 pseudovirus production The day before transfection, HEK293T/17 were seeded in a 15-cm dish at a density of 5 × 106 cells in DMEM supplemented with 10% heat inactivated FBS, non-essential amino acids, HEPES, and glutamine . .. Next day, cells were transfected with a mix of packaging vector (pALDI-Lenti System, Aldevron), luciferase reporter vector, and a plasmid encoding for the wild-type SARS-CoV2 Spike protein (VG40589-UT, Sino Biological) or vesicular stomatitis virus G (VSV-G, Aldevron), using FuGENE6 (Roche) as a transfection reagent:DNA ratio of 3:1, according to manufacturer’s protocol. .. Sixteen hours post-transfection, the media was replaced and cells were incubated for an additional 24–72 h. Supernatants were harvested at 24, 48, and 72 h, clarified by centrifugation at 1500 r.p.m. for 5 min and filtered using a sterile 0.22-µm pore size filter.

    Article Title: Real-Time Conformational Dynamics of SARS-CoV-2 Spikes on Virus Particles
    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. .. The translated amino acid sequence of pCMV-S is identical to QHD43416.1 (GenBank).

    Polymerase Chain Reaction:

    Article Title: Naturally mutated spike proteins of SARS-CoV-2 variants show differential levels of cell entry
    Article Snippet: The SARS-CoV S expression plasmid pC-SARS-S was created by inserting the BsiWI/XhoI-digested PCR-amplified SARS-CoV S fragment of CMV/R-SARS-S into the corresponding site of pCAGGS. .. The SARS-CoV-2 S expression plasmid pC-SARS2-S was created by inserting the Acc65I/NotI-digested PCR-amplified SARS-CoV-2 S fragment of pCMV3-2019-nCoV-Spike(S1+S2)-long (Sino Biological; VG40589-UT) into the corresponding site of pCAGGS. .. The SARS-CoV-2 S mutants (pC-SARS2-S-H49Y, pC-SARS2-S-V367F, pC-SARS2-S-G476S, pC-SARS2-S-V483A, pC-SARS2-S-D614G, or pC-SARS2-S-C1247A), in which positions 49, 367, 476, 483, 614, or 1247 of the S protein were mutated from histidine to tyrosine, valine to phenylalanine, glycine to serine, valine to alanine, aspartic acid to glycine, or cysteine to alanine, respectively, were created by inserting overlapping PCR fragments into Acc65I/NotI-digested pCAGGS.

    Recombinant:

    Article Title: SARS-CoV-2 and SARS-CoV Spike-RBD Structure and Receptor Binding Comparison and Potential Implications on Neutralizing Antibody and Vaccine Development
    Article Snippet: .. Reagents, recombinant proteins and antibodiesRecombinant 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. ..

    Luciferase:

    Article Title: Development of a multi-antigenic SARS-CoV-2 vaccine candidate using a synthetic poxvirus platform
    Article Snippet: SARS-CoV-2 pseudovirus production The day before transfection, HEK293T/17 were seeded in a 15-cm dish at a density of 5 × 106 cells in DMEM supplemented with 10% heat inactivated FBS, non-essential amino acids, HEPES, and glutamine . .. Next day, cells were transfected with a mix of packaging vector (pALDI-Lenti System, Aldevron), luciferase reporter vector, and a plasmid encoding for the wild-type SARS-CoV2 Spike protein (VG40589-UT, Sino Biological) or vesicular stomatitis virus G (VSV-G, Aldevron), using FuGENE6 (Roche) as a transfection reagent:DNA ratio of 3:1, according to manufacturer’s protocol. .. Sixteen hours post-transfection, the media was replaced and cells were incubated for an additional 24–72 h. Supernatants were harvested at 24, 48, and 72 h, clarified by centrifugation at 1500 r.p.m. for 5 min and filtered using a sterile 0.22-µm pore size filter.

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  • 98
    Sino Biological sars cov 2 2019 ncov spike orf mammalian expression plasmid codon optimized covid 19 spike research
    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 <t>SARS-CoV-2</t> 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., <t>2019</t> ). 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
    Sars Cov 2 2019 Ncov Spike Orf Mammalian Expression Plasmid Codon Optimized Covid 19 Spike Research, supplied by Sino Biological, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sars cov 2 2019 ncov spike orf mammalian expression plasmid codon optimized covid 19 spike research/product/Sino Biological
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 2019 ncov spike orf mammalian expression plasmid codon optimized covid 19 spike research - by Bioz Stars, 2021-04
    98/100 stars
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    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

    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

    Characterization of human anti-SARS-2 RBD Fabs. ( a ) Soluble ELISA of ten serially diluted human anti-SARS-2 RBD Fabs on immobilized SARS-2 RBD surfaces to measure their apparent affinities ( EC 50 , nM). ( b ) Schematic drawings of a competitive ELISA of human anti-SARS-2 RBD Fabs between the SARS-2 RBD and ACE2 protein (left) or ACE2-overexpressed cells (right). ( c ) Competitive ELISA of human anti-SARS-2 RBD Fabs antagonizing the interaction between ACE2 and the SARS-CoV-2 RBD. ( d ) Competitive flow cytometry analysis of human anti-SARS-2 RBD Fabs antagonizing the interaction between ACE2 on cells and the SARS-CoV-2 RBD (tagged with mouse Fc (mFc)). Arrows indicate potentially neutralizing clones. mFc-PE: anti-mouse PE (phycoerythrin) conjugate; MFI: mean fluorescence intensity; n.s: not significant ( p > 0.05); NC: negative control. * and **: p

    Journal: International Journal of Molecular Sciences

    Article Title: Neutralizing Human Antibodies against Severe Acute Respiratory Syndrome Coronavirus 2 Isolated from a Human Synthetic Fab Phage Display Library

    doi: 10.3390/ijms22041913

    Figure Lengend Snippet: Characterization of human anti-SARS-2 RBD Fabs. ( a ) Soluble ELISA of ten serially diluted human anti-SARS-2 RBD Fabs on immobilized SARS-2 RBD surfaces to measure their apparent affinities ( EC 50 , nM). ( b ) Schematic drawings of a competitive ELISA of human anti-SARS-2 RBD Fabs between the SARS-2 RBD and ACE2 protein (left) or ACE2-overexpressed cells (right). ( c ) Competitive ELISA of human anti-SARS-2 RBD Fabs antagonizing the interaction between ACE2 and the SARS-CoV-2 RBD. ( d ) Competitive flow cytometry analysis of human anti-SARS-2 RBD Fabs antagonizing the interaction between ACE2 on cells and the SARS-CoV-2 RBD (tagged with mouse Fc (mFc)). Arrows indicate potentially neutralizing clones. mFc-PE: anti-mouse PE (phycoerythrin) conjugate; MFI: mean fluorescence intensity; n.s: not significant ( p > 0.05); NC: negative control. * and **: p

    Article Snippet: Production of SARS-CoV-2 Spike pseudovirusPlasmids encoding the SARS-CoV-2 spike protein (D614) were purchased from Sino Biological (pCMV3-SARS-CoV-2 Spike, Cat. VG40589-UT, Beijing, China).

    Techniques: Enzyme-linked Immunosorbent Assay, Competitive ELISA, Flow Cytometry, Clone Assay, Fluorescence, Negative Control

    Characterization of anti-SARS-2 RBD immunoglobulin Gs (IgGs). ( a ) Binding analysis of five human anti-SARS-2 RBD IgGs—C12 (IgG), H1 (IgG), C2 (IgG), D12 (IgG), and F7 (IgG)—to the SARS-2 RBD and its variants (top) and the SARS-CoV-2 S1 (D614G) and other coronavirus S1 proteins (bottom), respectively. ( b ) Soluble ELISA of five serially diluted human anti-SARS-2 RBD IgGs on immobilized SARS-2 RBD surfaces to measure their apparent affinities ( EC 50 , nM). ( c ) ELISA detection for five human anti-SARS-2 RBD IgGs blocking the binding of the ACE2 protein with the SARS-CoV-2 RBD (top) and analysis of the flow cytometry for the blocking effect between the SARS-CoV-2 RBD and an ACE2-overexpressed cell (bottom). ( d ) Size-exclusion chromatography analysis of five human anti-SARS-2 RBD IgGs. The positions of the molecular mass markers, shown as kDa, on the retention time x -axis are indicated above the peaks. The data are presented as the mean ± standard error (SEM). MFI: mean fluorescence intensity; NC: negative control; *, **, and ***: p

    Journal: International Journal of Molecular Sciences

    Article Title: Neutralizing Human Antibodies against Severe Acute Respiratory Syndrome Coronavirus 2 Isolated from a Human Synthetic Fab Phage Display Library

    doi: 10.3390/ijms22041913

    Figure Lengend Snippet: Characterization of anti-SARS-2 RBD immunoglobulin Gs (IgGs). ( a ) Binding analysis of five human anti-SARS-2 RBD IgGs—C12 (IgG), H1 (IgG), C2 (IgG), D12 (IgG), and F7 (IgG)—to the SARS-2 RBD and its variants (top) and the SARS-CoV-2 S1 (D614G) and other coronavirus S1 proteins (bottom), respectively. ( b ) Soluble ELISA of five serially diluted human anti-SARS-2 RBD IgGs on immobilized SARS-2 RBD surfaces to measure their apparent affinities ( EC 50 , nM). ( c ) ELISA detection for five human anti-SARS-2 RBD IgGs blocking the binding of the ACE2 protein with the SARS-CoV-2 RBD (top) and analysis of the flow cytometry for the blocking effect between the SARS-CoV-2 RBD and an ACE2-overexpressed cell (bottom). ( d ) Size-exclusion chromatography analysis of five human anti-SARS-2 RBD IgGs. The positions of the molecular mass markers, shown as kDa, on the retention time x -axis are indicated above the peaks. The data are presented as the mean ± standard error (SEM). MFI: mean fluorescence intensity; NC: negative control; *, **, and ***: p

    Article Snippet: Production of SARS-CoV-2 Spike pseudovirusPlasmids encoding the SARS-CoV-2 spike protein (D614) were purchased from Sino Biological (pCMV3-SARS-CoV-2 Spike, Cat. VG40589-UT, Beijing, China).

    Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Blocking Assay, Flow Cytometry, Size-exclusion Chromatography, Fluorescence, Negative Control

    In vitro neutralization assay of human anti-SARS-2 RBD IgGs. Pseudo-typed virus-based neutralization ( a ) and a neutralization assay using authentic SARS-CoV-2 ( b ). ( c ) Correlation in neutralization potencies between pseudo-typed virus- and authentic virus-based assays. ( d ) Correlation between affinities of anti-SARS-2 RBD IgGs and their neutralization potencies for the authentic virus. The data are showed as the mean ± standard error (SEM).

    Journal: International Journal of Molecular Sciences

    Article Title: Neutralizing Human Antibodies against Severe Acute Respiratory Syndrome Coronavirus 2 Isolated from a Human Synthetic Fab Phage Display Library

    doi: 10.3390/ijms22041913

    Figure Lengend Snippet: In vitro neutralization assay of human anti-SARS-2 RBD IgGs. Pseudo-typed virus-based neutralization ( a ) and a neutralization assay using authentic SARS-CoV-2 ( b ). ( c ) Correlation in neutralization potencies between pseudo-typed virus- and authentic virus-based assays. ( d ) Correlation between affinities of anti-SARS-2 RBD IgGs and their neutralization potencies for the authentic virus. The data are showed as the mean ± standard error (SEM).

    Article Snippet: Production of SARS-CoV-2 Spike pseudovirusPlasmids encoding the SARS-CoV-2 spike protein (D614) were purchased from Sino Biological (pCMV3-SARS-CoV-2 Spike, Cat. VG40589-UT, Beijing, China).

    Techniques: In Vitro, Neutralization