depletion  (Sino Biological)


Bioz Verified Symbol Sino Biological is a verified supplier
Bioz Manufacturer Symbol Sino Biological manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Name:
    SARS CoV 2 2019 nCoV Spike RBD His Recombinant Protein COVID 19 Spike RBD Research
    Description:
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Spike Protein RBD YP 009724390 1 Arg319 Phe541 was expressed with a polyhistidine tag at the C terminus
    Catalog Number:
    40592-V08B
    Price:
    None
    Category:
    recombinant protein
    Product Aliases:
    coronavirus spike Protein 2019-nCoV, cov spike Protein 2019-nCoV, ncov RBD Protein 2019-nCoV, ncov s1 Protein 2019-nCoV, ncov s2 Protein 2019-nCoV, ncov spike Protein 2019-nCoV, NCP-CoV RBD Protein 2019-nCoV, NCP-CoV s1 Protein 2019-nCoV, NCP-CoV s2 Protein 2019-nCoV, NCP-CoV Spike Protein 2019-nCoV, novel coronavirus RBD Protein 2019-nCoV, novel coronavirus s1 Protein 2019-nCoV, novel coronavirus s2 Protein 2019-nCoV, novel coronavirus spike Protein 2019-nCoV, RBD Protein 2019-nCoV, S1 Protein 2019-nCoV, S2 Protein 2019-nCoV, Spike RBD Protein 2019-nCoV
    Host:
    Baculovirus-Insect Cells
    Buy from Supplier


    Structured Review

    Sino Biological depletion
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Spike Protein RBD YP 009724390 1 Arg319 Phe541 was expressed with a polyhistidine tag at the C terminus
    https://www.bioz.com/result/depletion/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    depletion - by Bioz Stars, 2021-04
    94/100 stars

    Images

    Related Articles

    Enzyme-linked Immunosorbent Assay:

    Article Title: Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19
    Article Snippet: .. Enzyme-linked immunosorbent assay As previously described, 50 ng of SARS-CoV-2 S1 protein (Sino Biological, 40591-V08H) or SARS-CoV-2 RBD protein (Sino Biological, 40592-V08B) or SARS-CoV-2 S2 protein (Sino Biological, 40590-V08B) in 100 μl phosphate-buffered saline (PBS) per well was coated on ELISA plates (Costar, 42592) overnight at 4 °C. .. The ELISA plates were blocked for 1 h with 100 μl blocking buffer (5% fetal bovine serum (FBS) and 0.1% Tween 20 in PBS) and then incubated with diluted patient or healthy control sera in 100 μl blocking buffer for 1 h. After washing with PBST buffer (0.1% Tween 20 in PBS), the ELISA plates were incubated with anti-human IgG horseradish peroxidase (HRP) antibody (Bioss Biotech, 0297D) for 45 min, followed by PBST washing and addition of 3,3′,5,5′-tetramethylbenzidine (TMB) buffer (Beyotime, P0209).

    Article Title: Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19
    Article Snippet: No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. strictly(RBD(dependent(and(deletion(of(RBD_specific(antibodies( led( to( the( failure( in( neutralization(in(S1_specific(sera. (These(results(highlighted(the(importance(of(S1_RBD( itself,(but(not(other(parts(of(S1(protein,(in(inducing(competent(NAbs. ( ( ( In(conclusion,(we(have(demonstrated(the(positive(correlation(between(the(magnitude( of(NAb(responses(and(disease(severity(in(patients(recovered(from(COVID_19. (We(have( also( found( that( disease( severity( also( influences( the( neutralization( heterogeneity( of( SARS_CoV2_specific(antibodies. (Our(results(highlight(the(needs(to(include(mild_illness( and(asymptomatic(patients(for( future(vaccination,(and(also(suggest( the(collection(of( plasma(from(COVID_19(recovered(patients(should(be(restricted(to(those(with(moderate( to( severe( symptoms( for( passive( antibody( therapy. (Our( data( also( provide( important( rationale( for( exclusively( using( SARS_CoV_2( RBD( as( S1_immunogen( in( COVID_19( vaccine(regime. ( . .. MaterialsHuman(The(59(COVID_19( recovered(patients(enrolled( in( the(study(were( provided(written(informed(consent(and(from(different(sources. (The(sera(of(the(severe,( moderate(and(mild(patients(were(obtained(from(Guangzhou(Eighth(People’s(Hospital. ( The(sera(of(the(asymptomatic(patients(were(obtained(from(Chongqing(Public(Health( Medical(Center. (Healthy(control(subjects(were(4(adult(participants( in( the(study. (The( study(received(IRB(approvals(at(Guangzhou(Eighth(People’s(Hospital((KE202001134)( and(Chongqing(Public(Health(Medical(Center((2020_023_01_KY). ( ELISA. (As(previously(described15,(50(ng(of(SARS_CoV_2(S1(protein((Sino(Biological,( 40591_V08H)(or(SARS_CoV_2(RBD(protein((Sino(Biological,(40592_V08B)(or(SARS_ CoV_2(S2(protein((Sino(Biological,(40590_V08B)(in(100(μl(PBS(per(well(was(coated(on( ELISA(plates((Costar,(42592)(overnight(at(4oC. (The(ELISA(plates(were(blocked(for(1( hour( with( 100( μl( blocking( buffer( (5%( FBS( and( 0.1%( Tween( 20( in( PBS)( and( then( incubated(with(diluted(patient(or(healthy(control(sera(in(100(μl(blocking(buffer(for(1(hour. ( All rights reserved. ..

    Multiplex Assay:

    Article Title: SARS-CoV-2-specific antibody detection for sero-epidemiology: a multiplex analysis approach accounting for accurate seroprevalence.
    Article Snippet: Assay ProcedureThe steps in assay validation were similar to recently developed bead-based multiplex immunoassays for CMV, EBV, and RSV, with minor modifications as described below [16, 17]. .. For the multiplex bead-based immune assay the following antigens obtained from Sino Biological were used: SARS-CoV-2 monomeric spike S1 (40591-V08H), RBD (40592-V08B), and nucleoprotein (N) (40588-V08B). ..

    Article Title: SARS-CoV-2–Specific Antibody Detection for Seroepidemiology: A Multiplex Analysis Approach Accounting for Accurate Seroprevalence
    Article Snippet: Assay Procedure The steps in assay validation were similar to recently developed bead-based multiplex immunoassays for CMV, EBV, and RSV, with minor modifications as described below [ , ]. .. For the multiplex bead-based immune assay the following antigens obtained from Sino Biological were used: SARS-CoV-2 monomeric spike S1 (40591-V08H), RBD (40592-V08B), and nucleoprotein (N) (40588-V08B). ..

    Recombinant:

    Article Title: Synthesis of polystyrene-based fluorescent quantum dots nanolabel and its performance in H5N1 virus and SARS-CoV-2 antibody sensing
    Article Snippet: Fluoro-max fluorescent beads with europium chelate was provided from Thermo Fisher Scientific Inc. COOH-coated polystyrene (PS) microbeads (0.2 and 4 μm) were obtained from Huge Biotechnology Co., Ltd (China). .. SARS-CoV-2 (2019-nCoV) Spike RBD-His Recombinant Protein were obtained from Sino Biological (Beijing). .. Influenza A virus H5N1 HA (Hemagglutinin) antibody were purchased from Gene Tex.

    Incubation:

    Article Title: Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19
    Article Snippet: No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. after(the(mixture(incubation,(the(luciferase(activity(of(SARS_CoV_2(typed(pseudovirus_ infected(ACE2/293T(cells(were(measured(by(a(luciferase(reporter(assay(kit((Promega,( E1910). ( SARS0CoV02(sera(were(diluted( in(DMEM((40( fold_dilution)( and( mixed( with( an( equal( volume( of( 80_100( PFU( SARS_CoV_2( (EPI_ISL_444969)(for(1(h(at(37°C. (Serum_virus(mixture(were(then(added(to(the(Vero( E6(cell(monolayers(in(48_well(plates(and(incubated(at(37°C(in(5%(CO2(for(1(h.(After( removing(the(inocula,(plates(were(overlaid(with(culture(medium(and(cultured(at(37°C( for(48(h.(Subsequently,(viral(RNA(from(the(cultural(supernatants(was(extracted(and(the( viral(RNA(copies(were(determined(by(quantitative(PCR(according(to(the(viral(detection( kit’s(protocol((DAAN(Gene(Co.,(Ltd.(of(Sun(Yat_Sen(University). (All(experiments(related( to( authentic( viruses( were( performed( in( the( certified( BSL_3( facility( of( Sun( Yat_sen( University.(The(SARS_CoV_2(viral(RNA(fold(reduction(=(2(CT(value(of(sample(_(CT(value(of(mock). .. Depletion(Firstly,(SARS_CoV_2(S1( protein((Sino(Biological,(40591_V08H)(or(SARS_CoV_2(RBD(protein((Sino(Biological,( 40592_V08B)(or(SARS_CoV_2(S2(protein(was(conjugated(with(biotin(by(following(the( manufacture’s( protocol( (Thermo(Fisher(Scientific,(A39257). (Then,( biotin_conjugated( proteins(were(incubated(with(BeaverBeads(Mag(Streptavidin(Matrix((Beaver,(22305)( at(4oC(for(1.5(hours. (After(washing(with(PBS,(the(SARS_CoV_2(S(protein(coupled(beads( were( next( incubated( with( diluted( patient( sera( at( 4oC( for( 1.5( hours. ( Then,( the( supernatants(were(harvested(and(quality(controlled(by(ELISA(assays(for(SARS_CoV_ 2(S(proteins. ( Statistics. (The(SARS_CoV_2(antibody(titers(or(virus(neutralizing(function(of(the(sera( belonging(to(patients(with(different(severity(were(compared(with(the(one_way(ANOVA( test( (Tukey’s( multiple( comparisons( test). ( The( cutoff( value( in( each( pseudovirus( neutralizing(function(assay(was(determined(by(the(ROC(curve(analysis(and(was(of(the( All rights reserved. .. No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. highest(likelihood(ratio. (Correlations(between(different(SARS_CoV_2(antibody(titers(or( between(SARS_CoV_2(antibody(titers(and(pseudovirus(titers(or(between(SARS_CoV_2( antibody(titers(and(SARS_CoV_2(virus(titers(were(analyzed(using(Pearson’s(correlation( coefficient. (P( values( less( than( 0.05(were( defied( as( statistically( significant. ( Prism( 6( software(was(used(for(statistical(analysis.

    Article Title: Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19
    Article Snippet: No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. strictly(RBD(dependent(and(deletion(of(RBD_specific(antibodies( led( to( the( failure( in( neutralization(in(S1_specific(sera. (These(results(highlighted(the(importance(of(S1_RBD( itself,(but(not(other(parts(of(S1(protein,(in(inducing(competent(NAbs. ( ( ( In(conclusion,(we(have(demonstrated(the(positive(correlation(between(the(magnitude( of(NAb(responses(and(disease(severity(in(patients(recovered(from(COVID_19. (We(have( also( found( that( disease( severity( also( influences( the( neutralization( heterogeneity( of( SARS_CoV2_specific(antibodies. (Our(results(highlight(the(needs(to(include(mild_illness( and(asymptomatic(patients(for( future(vaccination,(and(also(suggest( the(collection(of( plasma(from(COVID_19(recovered(patients(should(be(restricted(to(those(with(moderate( to( severe( symptoms( for( passive( antibody( therapy. (Our( data( also( provide( important( rationale( for( exclusively( using( SARS_CoV_2( RBD( as( S1_immunogen( in( COVID_19( vaccine(regime. ( . .. MaterialsHuman(The(59(COVID_19( recovered(patients(enrolled( in( the(study(were( provided(written(informed(consent(and(from(different(sources. (The(sera(of(the(severe,( moderate(and(mild(patients(were(obtained(from(Guangzhou(Eighth(People’s(Hospital. ( The(sera(of(the(asymptomatic(patients(were(obtained(from(Chongqing(Public(Health( Medical(Center. (Healthy(control(subjects(were(4(adult(participants( in( the(study. (The( study(received(IRB(approvals(at(Guangzhou(Eighth(People’s(Hospital((KE202001134)( and(Chongqing(Public(Health(Medical(Center((2020_023_01_KY). ( ELISA. (As(previously(described15,(50(ng(of(SARS_CoV_2(S1(protein((Sino(Biological,( 40591_V08H)(or(SARS_CoV_2(RBD(protein((Sino(Biological,(40592_V08B)(or(SARS_ CoV_2(S2(protein((Sino(Biological,(40590_V08B)(in(100(μl(PBS(per(well(was(coated(on( ELISA(plates((Costar,(42592)(overnight(at(4oC. (The(ELISA(plates(were(blocked(for(1( hour( with( 100( μl( blocking( buffer( (5%( FBS( and( 0.1%( Tween( 20( in( PBS)( and( then( incubated(with(diluted(patient(or(healthy(control(sera(in(100(μl(blocking(buffer(for(1(hour. ( All rights reserved. ..

    Blocking Assay:

    Article Title: Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19
    Article Snippet: No reuse allowed without permission. (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. strictly(RBD(dependent(and(deletion(of(RBD_specific(antibodies( led( to( the( failure( in( neutralization(in(S1_specific(sera. (These(results(highlighted(the(importance(of(S1_RBD( itself,(but(not(other(parts(of(S1(protein,(in(inducing(competent(NAbs. ( ( ( In(conclusion,(we(have(demonstrated(the(positive(correlation(between(the(magnitude( of(NAb(responses(and(disease(severity(in(patients(recovered(from(COVID_19. (We(have( also( found( that( disease( severity( also( influences( the( neutralization( heterogeneity( of( SARS_CoV2_specific(antibodies. (Our(results(highlight(the(needs(to(include(mild_illness( and(asymptomatic(patients(for( future(vaccination,(and(also(suggest( the(collection(of( plasma(from(COVID_19(recovered(patients(should(be(restricted(to(those(with(moderate( to( severe( symptoms( for( passive( antibody( therapy. (Our( data( also( provide( important( rationale( for( exclusively( using( SARS_CoV_2( RBD( as( S1_immunogen( in( COVID_19( vaccine(regime. ( . .. MaterialsHuman(The(59(COVID_19( recovered(patients(enrolled( in( the(study(were( provided(written(informed(consent(and(from(different(sources. (The(sera(of(the(severe,( moderate(and(mild(patients(were(obtained(from(Guangzhou(Eighth(People’s(Hospital. ( The(sera(of(the(asymptomatic(patients(were(obtained(from(Chongqing(Public(Health( Medical(Center. (Healthy(control(subjects(were(4(adult(participants( in( the(study. (The( study(received(IRB(approvals(at(Guangzhou(Eighth(People’s(Hospital((KE202001134)( and(Chongqing(Public(Health(Medical(Center((2020_023_01_KY). ( ELISA. (As(previously(described15,(50(ng(of(SARS_CoV_2(S1(protein((Sino(Biological,( 40591_V08H)(or(SARS_CoV_2(RBD(protein((Sino(Biological,(40592_V08B)(or(SARS_ CoV_2(S2(protein((Sino(Biological,(40590_V08B)(in(100(μl(PBS(per(well(was(coated(on( ELISA(plates((Costar,(42592)(overnight(at(4oC. (The(ELISA(plates(were(blocked(for(1( hour( with( 100( μl( blocking( buffer( (5%( FBS( and( 0.1%( Tween( 20( in( PBS)( and( then( incubated(with(diluted(patient(or(healthy(control(sera(in(100(μl(blocking(buffer(for(1(hour. ( All rights reserved. ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Sino Biological sars cov 2 2019 ncov spike rbd his recombinant protein covid 19 spike rbd research
    Laboratory confirmation of ACE2-based LFIA using clinical samples a) Schematic diagram of <t>COVID-19</t> test using ACE2-based LFIA. A nasopharyngeal swab from the COVID-19 patient is placed into the UTM. 50 μL of UTM containing the <t>SARS-CoV-2</t> is mixed with running buffer in a 1:1 (v/v) ratio, and 100 μL of mixed solution is loaded into the LFIA device. After 20 min, the line intensity of the LFIA strip is semi-quantified by the portable analyzer. b) Results of ACE2-based LFA for the detection sensitivity of cultured SARS-CoV-2. Serially diluted virus concentrates (concentration range: 1.07 × 10 8 copies/mL to 5.35 × 10 6 copies/mL) were tested. After 20 min, the LFIA strip was taken with a smartphone and scanned with an image analyzer. The line intensities of the test and control lines were converted to peak histograms. Also, the intensity of the test lines was measured by a portable line analyzer (I L : line intensity of test line). Furthermore, human coronavirus (OC43) was tested as a negative control. c) Bar graph of intensities for test lines measured by the portable analyzer. The limit of detection (LOD) was determined by the mean value of negative controls (0 copies/mL of SARS-CoV-2) plus three times the standard deviation. d) Laboratory confirmation of ACE2-based LFIA compared to the RT-qPCR using clinical samples. i) Nasopharyngeal swab samples of COVID-19 patients (n = 4) and healthy subjects (n = 4) were tested both ACE2-based LFIA and RT-qPCR. Sensitivity was determined by the number of true positive samples divided by the number of positive samples tested. Moreover, specificity was determined by the number of true negative samples divide by the number of negative samples tested. ii) RT-qPCR results for the detection of the SARS-CoV-2 specific gene (Env gene). C t value and their correspondent viral load in the clinical samples were evaluated. e) Results of ACE2-based LFIA on laboratory confirmation using clinical COVID-19 patient samples. Twenty minutes after sample loading, the test line intensities of the LFIA strips were measured with a portable line analyzer. The limit of detection (LOD) was determined by the mean value of negative controls (healthy control) plus three times the standard deviation.
    Sars Cov 2 2019 Ncov Spike Rbd His Recombinant Protein Covid 19 Spike Rbd Research, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, 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 rbd his recombinant protein covid 19 spike rbd research/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 2019 ncov spike rbd his recombinant protein covid 19 spike rbd research - by Bioz Stars, 2021-04
    94/100 stars
      Buy from Supplier

    Image Search Results


    Laboratory confirmation of ACE2-based LFIA using clinical samples a) Schematic diagram of COVID-19 test using ACE2-based LFIA. A nasopharyngeal swab from the COVID-19 patient is placed into the UTM. 50 μL of UTM containing the SARS-CoV-2 is mixed with running buffer in a 1:1 (v/v) ratio, and 100 μL of mixed solution is loaded into the LFIA device. After 20 min, the line intensity of the LFIA strip is semi-quantified by the portable analyzer. b) Results of ACE2-based LFA for the detection sensitivity of cultured SARS-CoV-2. Serially diluted virus concentrates (concentration range: 1.07 × 10 8 copies/mL to 5.35 × 10 6 copies/mL) were tested. After 20 min, the LFIA strip was taken with a smartphone and scanned with an image analyzer. The line intensities of the test and control lines were converted to peak histograms. Also, the intensity of the test lines was measured by a portable line analyzer (I L : line intensity of test line). Furthermore, human coronavirus (OC43) was tested as a negative control. c) Bar graph of intensities for test lines measured by the portable analyzer. The limit of detection (LOD) was determined by the mean value of negative controls (0 copies/mL of SARS-CoV-2) plus three times the standard deviation. d) Laboratory confirmation of ACE2-based LFIA compared to the RT-qPCR using clinical samples. i) Nasopharyngeal swab samples of COVID-19 patients (n = 4) and healthy subjects (n = 4) were tested both ACE2-based LFIA and RT-qPCR. Sensitivity was determined by the number of true positive samples divided by the number of positive samples tested. Moreover, specificity was determined by the number of true negative samples divide by the number of negative samples tested. ii) RT-qPCR results for the detection of the SARS-CoV-2 specific gene (Env gene). C t value and their correspondent viral load in the clinical samples were evaluated. e) Results of ACE2-based LFIA on laboratory confirmation using clinical COVID-19 patient samples. Twenty minutes after sample loading, the test line intensities of the LFIA strips were measured with a portable line analyzer. The limit of detection (LOD) was determined by the mean value of negative controls (healthy control) plus three times the standard deviation.

    Journal: Biosensors & Bioelectronics

    Article Title: A novel rapid detection for SARS-CoV-2 spike 1 antigens using human angiotensin converting enzyme 2 (ACE2)

    doi: 10.1016/j.bios.2020.112715

    Figure Lengend Snippet: Laboratory confirmation of ACE2-based LFIA using clinical samples a) Schematic diagram of COVID-19 test using ACE2-based LFIA. A nasopharyngeal swab from the COVID-19 patient is placed into the UTM. 50 μL of UTM containing the SARS-CoV-2 is mixed with running buffer in a 1:1 (v/v) ratio, and 100 μL of mixed solution is loaded into the LFIA device. After 20 min, the line intensity of the LFIA strip is semi-quantified by the portable analyzer. b) Results of ACE2-based LFA for the detection sensitivity of cultured SARS-CoV-2. Serially diluted virus concentrates (concentration range: 1.07 × 10 8 copies/mL to 5.35 × 10 6 copies/mL) were tested. After 20 min, the LFIA strip was taken with a smartphone and scanned with an image analyzer. The line intensities of the test and control lines were converted to peak histograms. Also, the intensity of the test lines was measured by a portable line analyzer (I L : line intensity of test line). Furthermore, human coronavirus (OC43) was tested as a negative control. c) Bar graph of intensities for test lines measured by the portable analyzer. The limit of detection (LOD) was determined by the mean value of negative controls (0 copies/mL of SARS-CoV-2) plus three times the standard deviation. d) Laboratory confirmation of ACE2-based LFIA compared to the RT-qPCR using clinical samples. i) Nasopharyngeal swab samples of COVID-19 patients (n = 4) and healthy subjects (n = 4) were tested both ACE2-based LFIA and RT-qPCR. Sensitivity was determined by the number of true positive samples divided by the number of positive samples tested. Moreover, specificity was determined by the number of true negative samples divide by the number of negative samples tested. ii) RT-qPCR results for the detection of the SARS-CoV-2 specific gene (Env gene). C t value and their correspondent viral load in the clinical samples were evaluated. e) Results of ACE2-based LFIA on laboratory confirmation using clinical COVID-19 patient samples. Twenty minutes after sample loading, the test line intensities of the LFIA strips were measured with a portable line analyzer. The limit of detection (LOD) was determined by the mean value of negative controls (healthy control) plus three times the standard deviation.

    Article Snippet: 2.2 Biolayer interferometryFc-tag tagged human ACE2 (ACE2, Cat. No. 10108-H05H), SARS-CoV-2 spike monoclonal antibody (S1-mAb, Cat. No. 40150-R007), SARS-CoV-2 spike protein (S1 subunit, Cat. No. 40591-V08H), and SARS-CoV-2 RBD (Cat. No. 40592-V08B) were purchased from Sino Biological.

    Techniques: Stripping Membranes, Cell Culture, Concentration Assay, Negative Control, Standard Deviation, Quantitative RT-PCR

    NP-based inhibition assay. (a) Left: The structure of neutralizing antibody (top) bound to SARS-CoV-2 Spike RBD (bottom, green). Right: Schematic diagram of the inhibition assay depicting blocking of the interaction between RBD and ACE2 and the resulting inhibition of energy transfer from QD to AuNP. (b) PL recovery of QD 514 -RBD in the presence of neutralizing antibody Ab1. (c) Inhibition test using anti-Spike antibody without neutralizing ability, showing almost no PL recovery of QD 514 -RBD. (d) Calculated EC 50 s for neutralizing antibodies Ab1 and Ab2 were 60 nM and 125 nM with R 2 > 99%, respectively.

    Journal: ACS Nano

    Article Title: Quantum Dot-Conjugated SARS-CoV-2 Spike Pseudo-Virions Enable Tracking of Angiotensin Converting Enzyme 2 Binding and Endocytosis

    doi: 10.1021/acsnano.0c05975

    Figure Lengend Snippet: NP-based inhibition assay. (a) Left: The structure of neutralizing antibody (top) bound to SARS-CoV-2 Spike RBD (bottom, green). Right: Schematic diagram of the inhibition assay depicting blocking of the interaction between RBD and ACE2 and the resulting inhibition of energy transfer from QD to AuNP. (b) PL recovery of QD 514 -RBD in the presence of neutralizing antibody Ab1. (c) Inhibition test using anti-Spike antibody without neutralizing ability, showing almost no PL recovery of QD 514 -RBD. (d) Calculated EC 50 s for neutralizing antibodies Ab1 and Ab2 were 60 nM and 125 nM with R 2 > 99%, respectively.

    Article Snippet: SARS-CoV S1-His (40150-V08B1), SARS-CoV-2 S1S2 ECD-His (40589-V08B1), SARS-CoV-2 S1-His (40591-V08H), SARS-CoV-2 RBD-His (40592-V08B), anti-SARS-CoV-2 S1 neutralizing antibody mouse mAb Ab1 (40591-MM43), and anti-SARS-CoV-2 RBD neutralizing antibody mouse mAb Ab2 (40592-MM57) were purchased from Sino Biological.

    Techniques: Inhibition, Blocking Assay

    Neutralizing antibody responses to SARS-CoV-2 in COVID-19 recovered patients. a Scores showing the COVID-19 patient serum-mediated inhibition of the SARS-CoV-2 RBD protein binding to ACE2 protein by ELISA. b Pie charts showing the proportions of patients with high ( > 50, green) or low (

    Journal: Signal Transduction and Targeted Therapy

    Article Title: Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19

    doi: 10.1038/s41392-020-00301-9

    Figure Lengend Snippet: Neutralizing antibody responses to SARS-CoV-2 in COVID-19 recovered patients. a Scores showing the COVID-19 patient serum-mediated inhibition of the SARS-CoV-2 RBD protein binding to ACE2 protein by ELISA. b Pie charts showing the proportions of patients with high ( > 50, green) or low (

    Article Snippet: Depletion of SARS-CoV-2 S protein-specific antibodiesFirst, SARS-CoV-2 S1 protein (Sino Biological, 40591-V08H) or SARS-CoV-2 RBD protein (Sino Biological, 40592-V08B) or SARS-CoV-2 S2 protein was conjugated with biotin by following the manufacturer’s protocol (Thermo Fisher Scientific, A39257).

    Techniques: Inhibition, Protein Binding, Enzyme-linked Immunosorbent Assay

    Subtypes of neutralizing antibodies to SARS-CoV-2 S proteins in COVID-19 recovered patients. a Blocking of luciferase-encoding SARS-CoV-2 typed pseudovirus into ACE2/293T cells by patient sera (no depletion) or S1 antibody-depleted sera (S1-Abs depletion) or S2 antibody-depleted sera (S2-Abs depletion). The dashed line indicates the cutoff value (6.749) determined by the ROC curve analysis. HC healthy control, NC negative control. b , c Pie charts showing the proportions of patients with different neutralizing antibody (NAb) subtype responses in the total 25 patients ( b ), 8 severe patients ( c , left panel), and 17 moderate and mild patients ( c , right panel) of pseudovirus neutralization positive. d Blocking of luciferase-encoding SARS-CoV-2 typed pseudovirus into ACE2/293T cells by “S1-NAbs only” patient sera with RBD antibody depletion (RBD-Abs depletion) or without RBD antibody depletion (no depletion). The dashed line indicates the cutoff value (6.034) determined by the ROC curve analysis. HC healthy control, NC negative control. e Pie chart showing the proportions of “S1-NAbs only” patients with RBD-Nab-dependent or -independent antibody response. Error bars in a , d indicate SEM

    Journal: Signal Transduction and Targeted Therapy

    Article Title: Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19

    doi: 10.1038/s41392-020-00301-9

    Figure Lengend Snippet: Subtypes of neutralizing antibodies to SARS-CoV-2 S proteins in COVID-19 recovered patients. a Blocking of luciferase-encoding SARS-CoV-2 typed pseudovirus into ACE2/293T cells by patient sera (no depletion) or S1 antibody-depleted sera (S1-Abs depletion) or S2 antibody-depleted sera (S2-Abs depletion). The dashed line indicates the cutoff value (6.749) determined by the ROC curve analysis. HC healthy control, NC negative control. b , c Pie charts showing the proportions of patients with different neutralizing antibody (NAb) subtype responses in the total 25 patients ( b ), 8 severe patients ( c , left panel), and 17 moderate and mild patients ( c , right panel) of pseudovirus neutralization positive. d Blocking of luciferase-encoding SARS-CoV-2 typed pseudovirus into ACE2/293T cells by “S1-NAbs only” patient sera with RBD antibody depletion (RBD-Abs depletion) or without RBD antibody depletion (no depletion). The dashed line indicates the cutoff value (6.034) determined by the ROC curve analysis. HC healthy control, NC negative control. e Pie chart showing the proportions of “S1-NAbs only” patients with RBD-Nab-dependent or -independent antibody response. Error bars in a , d indicate SEM

    Article Snippet: Depletion of SARS-CoV-2 S protein-specific antibodiesFirst, SARS-CoV-2 S1 protein (Sino Biological, 40591-V08H) or SARS-CoV-2 RBD protein (Sino Biological, 40592-V08B) or SARS-CoV-2 S2 protein was conjugated with biotin by following the manufacturer’s protocol (Thermo Fisher Scientific, A39257).

    Techniques: Blocking Assay, Luciferase, Negative Control, Neutralization

    Antibody responses to SARS-CoV-2 in COVID-19 recovered patients with different symptom severity. a – c ELISA binding assays of 100-fold diluted COVID-19 patient sera to ELISA plates after coating with SARS-CoV-2 S1 ( a ), RBD ( b ), and S2 ( c ) proteins. The dashed lines in a – c represent the average values of the healthy control groups. * P

    Journal: Signal Transduction and Targeted Therapy

    Article Title: Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19

    doi: 10.1038/s41392-020-00301-9

    Figure Lengend Snippet: Antibody responses to SARS-CoV-2 in COVID-19 recovered patients with different symptom severity. a – c ELISA binding assays of 100-fold diluted COVID-19 patient sera to ELISA plates after coating with SARS-CoV-2 S1 ( a ), RBD ( b ), and S2 ( c ) proteins. The dashed lines in a – c represent the average values of the healthy control groups. * P

    Article Snippet: Depletion of SARS-CoV-2 S protein-specific antibodiesFirst, SARS-CoV-2 S1 protein (Sino Biological, 40591-V08H) or SARS-CoV-2 RBD protein (Sino Biological, 40592-V08B) or SARS-CoV-2 S2 protein was conjugated with biotin by following the manufacturer’s protocol (Thermo Fisher Scientific, A39257).

    Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay

    Halofuginone Inhibits infection and replication of authentic SARS-CoV-2. a-b, Authentic SARS-CoV-2 virus infection of Huh7.5 cells treated with Halofuginone. Huh7.5 cells treated with halofuginone pre- or post-infection, or both pre- and post-infection with authentic SARS-CoV-2 virus. Viral titers ( a ) and quantification of viral RNA ( b ) in the infected cells is shown. c, Immunofluorescent quantification of viral nucleocapsid (red) protein in Vero E6 cells treated with Halofuginone and Remdesivir and infected with authentic SARS-CoV-2 virus (nuclei = green). d, Authentic SARS-CoV-2 virus infection of Vero E6 cells treated with Halofuginone and Remdesivir as measured in flow cytometry. e, Quantification of plaque formation and viral RNA in Vero E6 cells treated with Halofuginone and Remdesivir and infected with authentic SARS-CoV-2 virus. f, Rescue experiment of the effect of halofuginone treatment on SARS-CoV-2 infection using excess proline. g , Treatment of Vero E6 cells with halofuginone and enantiomers and their effect on SARS-CoV-2 infection. h, Treatment of Vero E6 cells with modulators of the PRS pathway and their effect on SARS-CoV-2 infection. i, Treatment of Vero E6 cells with modulators of the ISR pathway and their effect on SARS-CoV-2 infection. j, The distribution of proline distribution and density depicted as a proline distribution score for collagens, cholesterol biosynthetic proteins, lysosomal proteins and viral SARS-CoV-2 (SARS2), SARS-CoV-1 (SARS1), MERS-CoV (MERS), HCoV-229E and HCoV-LN63 proteins( ## p ≤ 0.01 collagen s vs. all other protein classes). k, Treatment of Vero E6 cells with AARS inhibitors and their effect on SARS-CoV-2 infection. Data shown as mean ± S.D. Statistics performed by 1-way ANOVA and uncorrected Fisher’s LSD test (ns: p > 0.05, *: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001, ****: p ≤ 0.0001, ## : p ≤ 0.01.

    Journal: bioRxiv

    Article Title: The Prolyl-tRNA Synthetase Inhibitor Halofuginone Inhibits SARS-CoV-2 Infection

    doi: 10.1101/2021.03.22.436522

    Figure Lengend Snippet: Halofuginone Inhibits infection and replication of authentic SARS-CoV-2. a-b, Authentic SARS-CoV-2 virus infection of Huh7.5 cells treated with Halofuginone. Huh7.5 cells treated with halofuginone pre- or post-infection, or both pre- and post-infection with authentic SARS-CoV-2 virus. Viral titers ( a ) and quantification of viral RNA ( b ) in the infected cells is shown. c, Immunofluorescent quantification of viral nucleocapsid (red) protein in Vero E6 cells treated with Halofuginone and Remdesivir and infected with authentic SARS-CoV-2 virus (nuclei = green). d, Authentic SARS-CoV-2 virus infection of Vero E6 cells treated with Halofuginone and Remdesivir as measured in flow cytometry. e, Quantification of plaque formation and viral RNA in Vero E6 cells treated with Halofuginone and Remdesivir and infected with authentic SARS-CoV-2 virus. f, Rescue experiment of the effect of halofuginone treatment on SARS-CoV-2 infection using excess proline. g , Treatment of Vero E6 cells with halofuginone and enantiomers and their effect on SARS-CoV-2 infection. h, Treatment of Vero E6 cells with modulators of the PRS pathway and their effect on SARS-CoV-2 infection. i, Treatment of Vero E6 cells with modulators of the ISR pathway and their effect on SARS-CoV-2 infection. j, The distribution of proline distribution and density depicted as a proline distribution score for collagens, cholesterol biosynthetic proteins, lysosomal proteins and viral SARS-CoV-2 (SARS2), SARS-CoV-1 (SARS1), MERS-CoV (MERS), HCoV-229E and HCoV-LN63 proteins( ## p ≤ 0.01 collagen s vs. all other protein classes). k, Treatment of Vero E6 cells with AARS inhibitors and their effect on SARS-CoV-2 infection. Data shown as mean ± S.D. Statistics performed by 1-way ANOVA and uncorrected Fisher’s LSD test (ns: p > 0.05, *: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001, ****: p ≤ 0.0001, ## : p ≤ 0.01.

    Article Snippet: Reagents SARS-CoV-2 (2019-nCoV) Spike Protein (RBD, His Tag) (Sino Biologicals, 40592-V08B), and proteins were biotinylated using EZ-Link™ Sulfo-NHS-Biotin, No-Weigh™ (Thermo Fisher Scientific, A39256).

    Techniques: Infection, Flow Cytometry

    Halofuginone Inhibition of heparan sulfate presentation and Spike protein binding is not dependent of the integrated stress response. a, Chemical structure of halofuginone and negative control compound MAZ1310. Figure shows the interaction of halofuginone with the human prolyl-tRNA synthetase (PRS) active site, as resolved by X-ray crystallography (PDB: 4K88) 48 . b, Chemical structure of ProSA. Graphic shows the interaction of ProSA with human PRS (PDB: 5V58) 49 . c, Schematic representation of the mediators of the integrated stress response (ISR). c-d, Treatment of Hep3B cells with modulators of the PRS pathway at 0.5 µM (ProSA at 5 µM) and its effect on ( c ) HS presentation as measured by anti-HS mAb 10E4 binding and ( d ) spike RBD binding ( e ) by flow cytometry. Binding is represented as relative to non-treated control. f, Treatment of Hep3B cells with modulators of the halofuginone with or without 4mM proline and its effect on spike RBD binding by flow cytometry. Binding is represented as relative to non-treated control. g, Treatment of Hep3B cells with modulators of the ISR and its effect on HS presentation as measured by anti-HS mAb 10E4 binding in flow cytometry. h, Treatment of Hep3B cells with modulators of the ISR and its effect on SARS-CoV-2 recombinant RBD protein binding in flow cytometry. Binding is represented as relative to non-treated control. i, The distribution of proline distribution and density depicted as a proline distribution score for collagens, cholesterol biosynthetic proteins, lysosomal proteins, heparan sulfate (HS) biosynthetic proteins, heparan sulfate proteoglycans (HSPG) and viral host factor proteins ( ## p ≤ 0.01 collagens vs. all other protein classes). Data shown as mean ± S.D. Statistics performed by 1-way ANOVA and uncorrected Fisher’s LSD test (ns: p > 0.05, *: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001, ****: p ≤ 0.0001; # : p ≤ 0.05, ## : p ≤ 0.01).

    Journal: bioRxiv

    Article Title: The Prolyl-tRNA Synthetase Inhibitor Halofuginone Inhibits SARS-CoV-2 Infection

    doi: 10.1101/2021.03.22.436522

    Figure Lengend Snippet: Halofuginone Inhibition of heparan sulfate presentation and Spike protein binding is not dependent of the integrated stress response. a, Chemical structure of halofuginone and negative control compound MAZ1310. Figure shows the interaction of halofuginone with the human prolyl-tRNA synthetase (PRS) active site, as resolved by X-ray crystallography (PDB: 4K88) 48 . b, Chemical structure of ProSA. Graphic shows the interaction of ProSA with human PRS (PDB: 5V58) 49 . c, Schematic representation of the mediators of the integrated stress response (ISR). c-d, Treatment of Hep3B cells with modulators of the PRS pathway at 0.5 µM (ProSA at 5 µM) and its effect on ( c ) HS presentation as measured by anti-HS mAb 10E4 binding and ( d ) spike RBD binding ( e ) by flow cytometry. Binding is represented as relative to non-treated control. f, Treatment of Hep3B cells with modulators of the halofuginone with or without 4mM proline and its effect on spike RBD binding by flow cytometry. Binding is represented as relative to non-treated control. g, Treatment of Hep3B cells with modulators of the ISR and its effect on HS presentation as measured by anti-HS mAb 10E4 binding in flow cytometry. h, Treatment of Hep3B cells with modulators of the ISR and its effect on SARS-CoV-2 recombinant RBD protein binding in flow cytometry. Binding is represented as relative to non-treated control. i, The distribution of proline distribution and density depicted as a proline distribution score for collagens, cholesterol biosynthetic proteins, lysosomal proteins, heparan sulfate (HS) biosynthetic proteins, heparan sulfate proteoglycans (HSPG) and viral host factor proteins ( ## p ≤ 0.01 collagens vs. all other protein classes). Data shown as mean ± S.D. Statistics performed by 1-way ANOVA and uncorrected Fisher’s LSD test (ns: p > 0.05, *: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001, ****: p ≤ 0.0001; # : p ≤ 0.05, ## : p ≤ 0.01).

    Article Snippet: Reagents SARS-CoV-2 (2019-nCoV) Spike Protein (RBD, His Tag) (Sino Biologicals, 40592-V08B), and proteins were biotinylated using EZ-Link™ Sulfo-NHS-Biotin, No-Weigh™ (Thermo Fisher Scientific, A39256).

    Techniques: Inhibition, Protein Binding, Negative Control, Binding Assay, Flow Cytometry, Recombinant

    Screen of epigenetic and translational regulatory compounds identify Halofuginone as a potent inhibitor of SARS-CoV-2 Spike HS dependent cellular adhesion. a, Hep3B cells were treated with a library of epigenetic and translational regulatory compounds or heparin lyase (HSase) as a positive control and tested for their interaction with recombinant SARS-CoV-2 RBD protein. b-e, Titration of halofuginone on Hep3B cells ( b ), Vero E6 ( c ), Calu-3 ( d ) and Cacu-2 ( e ) cells (n= 2-4 replicates per condition), and its effect on binding of recombinant SARS-CoV-2 RBD protein Heparin Lyase treatment is included as control for HS dependent adhesion. f-g, effect of halofuginone treatment on the infection of SARS-CoV-2 spike protein pseudotyped virus in Hep3B ( f ) and Vero E6 ( g ) cells (n= 3-4 replicates per condition). h , Authentic SARS-CoV-2 virus infection of Hep3B cells treated with halofuginone. i, Infection of primary human bronchial epithelial cells, grown at an air-liquid interface, treated with halofuginone (Red and black colors represent the two different primary human bronchial epithelial cells isolates used). j , Relative cell viability of primary human bronchial epithelial cells. Data shown as mean ± S.D. Statistics performed by 1-way ANOVA and uncorrected Fisher’s LSD test (ns: p > 0.05, *: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001).

    Journal: bioRxiv

    Article Title: The Prolyl-tRNA Synthetase Inhibitor Halofuginone Inhibits SARS-CoV-2 Infection

    doi: 10.1101/2021.03.22.436522

    Figure Lengend Snippet: Screen of epigenetic and translational regulatory compounds identify Halofuginone as a potent inhibitor of SARS-CoV-2 Spike HS dependent cellular adhesion. a, Hep3B cells were treated with a library of epigenetic and translational regulatory compounds or heparin lyase (HSase) as a positive control and tested for their interaction with recombinant SARS-CoV-2 RBD protein. b-e, Titration of halofuginone on Hep3B cells ( b ), Vero E6 ( c ), Calu-3 ( d ) and Cacu-2 ( e ) cells (n= 2-4 replicates per condition), and its effect on binding of recombinant SARS-CoV-2 RBD protein Heparin Lyase treatment is included as control for HS dependent adhesion. f-g, effect of halofuginone treatment on the infection of SARS-CoV-2 spike protein pseudotyped virus in Hep3B ( f ) and Vero E6 ( g ) cells (n= 3-4 replicates per condition). h , Authentic SARS-CoV-2 virus infection of Hep3B cells treated with halofuginone. i, Infection of primary human bronchial epithelial cells, grown at an air-liquid interface, treated with halofuginone (Red and black colors represent the two different primary human bronchial epithelial cells isolates used). j , Relative cell viability of primary human bronchial epithelial cells. Data shown as mean ± S.D. Statistics performed by 1-way ANOVA and uncorrected Fisher’s LSD test (ns: p > 0.05, *: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001).

    Article Snippet: Reagents SARS-CoV-2 (2019-nCoV) Spike Protein (RBD, His Tag) (Sino Biologicals, 40592-V08B), and proteins were biotinylated using EZ-Link™ Sulfo-NHS-Biotin, No-Weigh™ (Thermo Fisher Scientific, A39256).

    Techniques: Positive Control, Recombinant, Titration, Binding Assay, Infection