sars cov 2 2019 ncov nucleocapsid his recombinant protein covid 19 nucleocapsid research  (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
  • 96
    Name:
    SARS CoV 2 2019 nCoV Nucleocapsid His recombinant Protein COVID 19 Nucleocapsid Research
    Description:
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Nucleocapsid Protein YP 009724397 2 335Gly Ala Met1 Ala419 was expressed with a polyhistidine tag at the C terminus
    Catalog Number:
    40588-V08B
    Price:
    None
    Category:
    recombinant protein
    Product Aliases:
    coronavirus NP Protein 2019-nCoV, coronavirus Nucleocapsid Protein 2019-nCoV, coronavirus Nucleoprotein Protein 2019-nCoV, cov np Protein 2019-nCoV, ncov NP Protein 2019-nCoV, NCP-CoV Nucleocapsid Protein 2019-nCoV, novel coronavirus NP Protein 2019-nCoV, novel coronavirus Nucleocapsid Protein 2019-nCoV, novel coronavirus Nucleoprotein Protein 2019-nCoV, np Protein 2019-nCoV, nucleocapsid Protein 2019-nCoV, Nucleoprotein Protein 2019-nCoV
    Host:
    Baculovirus-Insect Cells
    Buy from Supplier


    Structured Review

    Sino Biological sars cov 2 2019 ncov nucleocapsid his recombinant protein covid 19 nucleocapsid research
    GCG suppresses <t>SARS-CoV-2</t> replication. a , b Immunofluorescence analysis of N protein in A549-hACE2-Flag cells infected with SARS-CoV-2 for 24 h ( a ). The percentage of cells with N protein foci was quantified, n = 8 biologically independent samples, 20 randomly selected views were analyzed in each sample ( b ). Scale bar, 10 μm. c 3D images were obtained by Zeiss LSM 880 confocal microscope and reconstituted by Volocity 6.1.1 . d , e The inhibitory effect of GCG on the replication of SARS-CoV-2, n = 6 biologically independent samples ( d ). IC 50 was calculated, n = 5 biologically independent samples ( e ). The infection was performed after 1-h pretreatment of GCG. f , g Representative immunofluorescent images showed the inhibitory effect of GCG on SARS-CoV-2 N protein. 3D images were obtained by Zeiss LSM 880 confocal microscope and reconstituted by Volocity 6.1.1 ( f ). Violin plots showing foci of cells ( n = 50 biologically independent cells) from each group, lines within the plots, with 25th, 50th, and 75th percentiles marked ( g ). h Cells were infected with SARS-CoV-2 for 1 h followed by 24-h GCG treatment, n = 3 biologically independent samples. Representative images were shown. SARS-CoV-2 was used at an MOI of 1. Hoechst (blue), nuclear staining ( a , c , f ). Error bars, mean with s.d. ( b , d , e , g , h ). Two-tailed unpaired Student’s t -test, * P
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Nucleocapsid Protein YP 009724397 2 335Gly Ala Met1 Ala419 was expressed with a polyhistidine tag at the C terminus
    https://www.bioz.com/result/sars cov 2 2019 ncov nucleocapsid his recombinant protein covid 19 nucleocapsid research/product/Sino Biological
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 2019 ncov nucleocapsid his recombinant protein covid 19 nucleocapsid research - by Bioz Stars, 2021-07
    96/100 stars

    Images

    1) Product Images from "GCG inhibits SARS-CoV-2 replication by disrupting the liquid phase condensation of its nucleocapsid protein"

    Article Title: GCG inhibits SARS-CoV-2 replication by disrupting the liquid phase condensation of its nucleocapsid protein

    Journal: Nature Communications

    doi: 10.1038/s41467-021-22297-8

    GCG suppresses SARS-CoV-2 replication. a , b Immunofluorescence analysis of N protein in A549-hACE2-Flag cells infected with SARS-CoV-2 for 24 h ( a ). The percentage of cells with N protein foci was quantified, n = 8 biologically independent samples, 20 randomly selected views were analyzed in each sample ( b ). Scale bar, 10 μm. c 3D images were obtained by Zeiss LSM 880 confocal microscope and reconstituted by Volocity 6.1.1 . d , e The inhibitory effect of GCG on the replication of SARS-CoV-2, n = 6 biologically independent samples ( d ). IC 50 was calculated, n = 5 biologically independent samples ( e ). The infection was performed after 1-h pretreatment of GCG. f , g Representative immunofluorescent images showed the inhibitory effect of GCG on SARS-CoV-2 N protein. 3D images were obtained by Zeiss LSM 880 confocal microscope and reconstituted by Volocity 6.1.1 ( f ). Violin plots showing foci of cells ( n = 50 biologically independent cells) from each group, lines within the plots, with 25th, 50th, and 75th percentiles marked ( g ). h Cells were infected with SARS-CoV-2 for 1 h followed by 24-h GCG treatment, n = 3 biologically independent samples. Representative images were shown. SARS-CoV-2 was used at an MOI of 1. Hoechst (blue), nuclear staining ( a , c , f ). Error bars, mean with s.d. ( b , d , e , g , h ). Two-tailed unpaired Student’s t -test, * P
    Figure Legend Snippet: GCG suppresses SARS-CoV-2 replication. a , b Immunofluorescence analysis of N protein in A549-hACE2-Flag cells infected with SARS-CoV-2 for 24 h ( a ). The percentage of cells with N protein foci was quantified, n = 8 biologically independent samples, 20 randomly selected views were analyzed in each sample ( b ). Scale bar, 10 μm. c 3D images were obtained by Zeiss LSM 880 confocal microscope and reconstituted by Volocity 6.1.1 . d , e The inhibitory effect of GCG on the replication of SARS-CoV-2, n = 6 biologically independent samples ( d ). IC 50 was calculated, n = 5 biologically independent samples ( e ). The infection was performed after 1-h pretreatment of GCG. f , g Representative immunofluorescent images showed the inhibitory effect of GCG on SARS-CoV-2 N protein. 3D images were obtained by Zeiss LSM 880 confocal microscope and reconstituted by Volocity 6.1.1 ( f ). Violin plots showing foci of cells ( n = 50 biologically independent cells) from each group, lines within the plots, with 25th, 50th, and 75th percentiles marked ( g ). h Cells were infected with SARS-CoV-2 for 1 h followed by 24-h GCG treatment, n = 3 biologically independent samples. Representative images were shown. SARS-CoV-2 was used at an MOI of 1. Hoechst (blue), nuclear staining ( a , c , f ). Error bars, mean with s.d. ( b , d , e , g , h ). Two-tailed unpaired Student’s t -test, * P

    Techniques Used: Immunofluorescence, Infection, Microscopy, Staining, Two Tailed Test

    RNA triggers the LLPS of N protein. a Schematic drawing of SARS-CoV-2. b IDR scores of 29 proteins encoded by SARS-CoV-2 genome. FUS and mEGFP are positive and negative controls, respectively. IUPred2 and ANCHOR2 were used as prediction tools. c Time-lapse imaging of N-mEGFP protein (20 μM) in the presence of Cy5-labeled 60-nt vRNA (100 ng/μl), scale bar, 10 μm. d Representative fluorescent images of N-mEGFP-vRNA (60 nt) condensates fusion from a time-lapse movie, scale bar, 3 μm. e – g LLPS of N-mEGFP protein (20 μM) in the presence of indicated concentrations of 60-nt vRNA, scale bar, 10 μm ( e ). The partition coefficient of fluorescence intensity per droplet ( f ) and the partition coefficient of total fluorescence intensity in each view ( g ) were calculated. From left to right, n = 209, 1170, 1026, 1170 droplets ( f ) from 10 randomly selected views ( g ). h , i FRAP analysis of vRNA-induced liquid droplets of N-mEGFP protein, scale bar, 2 μm ( h ), and quantification of fluorescence intensity recovery of a photobleached N-mEGFP protein, n = 3 biologically independent experiments ( i ). The white dotted circle in h indicated the region of photobleaching. 20 μM N-mEGFP protein and 100 ng/μl 60-nt vRNA were used. Error bars, mean with s.d. ( f , g , i ). Two-tailed unpaired Student’s t -test ( f , g ), **** P
    Figure Legend Snippet: RNA triggers the LLPS of N protein. a Schematic drawing of SARS-CoV-2. b IDR scores of 29 proteins encoded by SARS-CoV-2 genome. FUS and mEGFP are positive and negative controls, respectively. IUPred2 and ANCHOR2 were used as prediction tools. c Time-lapse imaging of N-mEGFP protein (20 μM) in the presence of Cy5-labeled 60-nt vRNA (100 ng/μl), scale bar, 10 μm. d Representative fluorescent images of N-mEGFP-vRNA (60 nt) condensates fusion from a time-lapse movie, scale bar, 3 μm. e – g LLPS of N-mEGFP protein (20 μM) in the presence of indicated concentrations of 60-nt vRNA, scale bar, 10 μm ( e ). The partition coefficient of fluorescence intensity per droplet ( f ) and the partition coefficient of total fluorescence intensity in each view ( g ) were calculated. From left to right, n = 209, 1170, 1026, 1170 droplets ( f ) from 10 randomly selected views ( g ). h , i FRAP analysis of vRNA-induced liquid droplets of N-mEGFP protein, scale bar, 2 μm ( h ), and quantification of fluorescence intensity recovery of a photobleached N-mEGFP protein, n = 3 biologically independent experiments ( i ). The white dotted circle in h indicated the region of photobleaching. 20 μM N-mEGFP protein and 100 ng/μl 60-nt vRNA were used. Error bars, mean with s.d. ( f , g , i ). Two-tailed unpaired Student’s t -test ( f , g ), **** P

    Techniques Used: Imaging, Labeling, Fluorescence, Two Tailed Test

    NR203K/G204R gained greater ability to undergo RNA-induced LLPS. a Distribution of N gene variants among 100,849 SARS-CoV-2 genomes obtained from GISAID database. Colors indicated the nucleotide variability numbers from 100,849 genomes. The high-frequency trio-nucleotide polymorphism variant (GGG-to-AAC) is shown. b Coomassie brilliant blue-stained SDS-PAGE gel of purified variants of N-mEGFP protein. c – e LLPS of different N-mEGFP variants, in the presence of 50 ng/μl 60-nt vRNA ( c ). The partition coefficient of fluorescence intensity per droplet ( d ) and the partition coefficient of total fluorescence intensity in each view ( e ) were calculated. From left to right, n = 1232, 803, 897, 431 droplets ( d ) from 10 randomly selected views ( e ). f , g Time-lapse imaging of N R203/G204 -mEGFP and N R203K/G204R -mEGFP proteins (20 μM) in the presence of Cy5-labeled 60-nt vRNA (40 ng/μl) ( f ), and the partition coefficient ( n = 8 randomly selected views) of total fluorescence intensity in each view ( g ). Scale bars, 10 μm ( c , f ). Error bars, mean with s.d. ( d , e ) and mean with s.e.m. ( g ). Two-tailed unpaired Student’s t -test ( d , e ), **** P
    Figure Legend Snippet: NR203K/G204R gained greater ability to undergo RNA-induced LLPS. a Distribution of N gene variants among 100,849 SARS-CoV-2 genomes obtained from GISAID database. Colors indicated the nucleotide variability numbers from 100,849 genomes. The high-frequency trio-nucleotide polymorphism variant (GGG-to-AAC) is shown. b Coomassie brilliant blue-stained SDS-PAGE gel of purified variants of N-mEGFP protein. c – e LLPS of different N-mEGFP variants, in the presence of 50 ng/μl 60-nt vRNA ( c ). The partition coefficient of fluorescence intensity per droplet ( d ) and the partition coefficient of total fluorescence intensity in each view ( e ) were calculated. From left to right, n = 1232, 803, 897, 431 droplets ( d ) from 10 randomly selected views ( e ). f , g Time-lapse imaging of N R203/G204 -mEGFP and N R203K/G204R -mEGFP proteins (20 μM) in the presence of Cy5-labeled 60-nt vRNA (40 ng/μl) ( f ), and the partition coefficient ( n = 8 randomly selected views) of total fluorescence intensity in each view ( g ). Scale bars, 10 μm ( c , f ). Error bars, mean with s.d. ( d , e ) and mean with s.e.m. ( g ). Two-tailed unpaired Student’s t -test ( d , e ), **** P

    Techniques Used: Variant Assay, Staining, SDS Page, Purification, Fluorescence, Imaging, Labeling, Two Tailed Test

    2) Product Images from "A SARS-CoV-2 antibody curbs viral nucleocapsid protein-induced complement hyperactivation"

    Article Title: A SARS-CoV-2 antibody curbs viral nucleocapsid protein-induced complement hyperactivation

    Journal: Nature Communications

    doi: 10.1038/s41467-021-23036-9

    Antibody nCoV396 compromises SARS-CoV-2 N protein-induced complement hyperactivation. a Flow scheme of the SARS-CoV-2 N protein and nCoV396 influencing the protease activity of MASP-2 in the serum from donors. b Serum-01 to 06 are used to compare the MASP-2 activity to C2 of serum sample with normal C3 (Health-110,113,117, n = 3) and serum sample with abnormal C3 (Patient-81,123,130, n = 3), and the Michaelis–Menten curves of are presented as mean (three groups of above health serum alone with patient’s serum paired data are shown in Supplementary Fig. 8a ). c The Michaelis–Menten curve of N protein-induced excessive cleavage of C2 in the presence of recombinant MASP-2 in vitro. The reaction system without N protein, the increase of N protein concentration and negative control protein (ENL) expressed in E. coli are presenting. The Michaelis–Menten curve shows the effect of increasing the N protein concentration ( d ) and antibody concentration ( f ) on the substrate C2 cleavage of MAPS-2 in the Serum-07 and Serum-08. e A Hanes plot where C2 concentration/V0 is plotted against C2 concentration with the addition of 5 μM N protein. b – d , f All samples were performed in triplicates and mean were presented. g Five serum sample from biologically independent donors ( n = 5) with abnormal serologic C3 values. (Serum-08 to −12). And we used Michaelis–Menten equation to calculate the V max (with experimental data from Fig. 4f (Serum-08), Supplementary Fig. 5b (Serum-09 to −11), and Supplementary Fig. 7a (Serum-12)). Each sample was performed in triplicates and mean values ± SEM of V max are presented. Two-sided Kruskal–Wallis test with Dunnett’s multiple comparisons test was used for comparing the V max of groups. The significant reference is 0.05.
    Figure Legend Snippet: Antibody nCoV396 compromises SARS-CoV-2 N protein-induced complement hyperactivation. a Flow scheme of the SARS-CoV-2 N protein and nCoV396 influencing the protease activity of MASP-2 in the serum from donors. b Serum-01 to 06 are used to compare the MASP-2 activity to C2 of serum sample with normal C3 (Health-110,113,117, n = 3) and serum sample with abnormal C3 (Patient-81,123,130, n = 3), and the Michaelis–Menten curves of are presented as mean (three groups of above health serum alone with patient’s serum paired data are shown in Supplementary Fig. 8a ). c The Michaelis–Menten curve of N protein-induced excessive cleavage of C2 in the presence of recombinant MASP-2 in vitro. The reaction system without N protein, the increase of N protein concentration and negative control protein (ENL) expressed in E. coli are presenting. The Michaelis–Menten curve shows the effect of increasing the N protein concentration ( d ) and antibody concentration ( f ) on the substrate C2 cleavage of MAPS-2 in the Serum-07 and Serum-08. e A Hanes plot where C2 concentration/V0 is plotted against C2 concentration with the addition of 5 μM N protein. b – d , f All samples were performed in triplicates and mean were presented. g Five serum sample from biologically independent donors ( n = 5) with abnormal serologic C3 values. (Serum-08 to −12). And we used Michaelis–Menten equation to calculate the V max (with experimental data from Fig. 4f (Serum-08), Supplementary Fig. 5b (Serum-09 to −11), and Supplementary Fig. 7a (Serum-12)). Each sample was performed in triplicates and mean values ± SEM of V max are presented. Two-sided Kruskal–Wallis test with Dunnett’s multiple comparisons test was used for comparing the V max of groups. The significant reference is 0.05.

    Techniques Used: Activity Assay, Recombinant, In Vitro, Protein Concentration, Negative Control, Concentration Assay

    Complex structure of mAb nCoV396 with SARS-CoV-2 N-NTD. a Overall structure of the mAb nCoV396-SARS-CoV-2 N-NTD complex. The light chain (pink) and heavy chain (blue) of mAb nCoV396 are illustrated with the ribbon representation. SARS-CoV-2 N-NTD is illustrated with electrostatics surface, in which blue denotes a positive charge potential while red indicates a negative charge potential. b The N-NTD epitope recognized by mAb nCoV396. The interacting residues of N-NTD and nCoV396 are highlighted with the stick representation. Recognition of Q163 ( c ), K169 ( d ), and L167 ( e ) in N-NTD by mAb nCoV396. The dashed blue line represents hydrogen bonds. Hydrophobic interactions are illustrated with the dot representation. f Conformational changes of N-NTD upon mAb nCoV396 binding. The apo structure of N-NTD is colored with gray. Antibody-bound N-NTD is colored green. The N-terminus and C-terminus of the N-NTD are labeled with circles. mAb nCoV396 is illustrated with surface representation. All figures were prepared by Pymol.
    Figure Legend Snippet: Complex structure of mAb nCoV396 with SARS-CoV-2 N-NTD. a Overall structure of the mAb nCoV396-SARS-CoV-2 N-NTD complex. The light chain (pink) and heavy chain (blue) of mAb nCoV396 are illustrated with the ribbon representation. SARS-CoV-2 N-NTD is illustrated with electrostatics surface, in which blue denotes a positive charge potential while red indicates a negative charge potential. b The N-NTD epitope recognized by mAb nCoV396. The interacting residues of N-NTD and nCoV396 are highlighted with the stick representation. Recognition of Q163 ( c ), K169 ( d ), and L167 ( e ) in N-NTD by mAb nCoV396. The dashed blue line represents hydrogen bonds. Hydrophobic interactions are illustrated with the dot representation. f Conformational changes of N-NTD upon mAb nCoV396 binding. The apo structure of N-NTD is colored with gray. Antibody-bound N-NTD is colored green. The N-terminus and C-terminus of the N-NTD are labeled with circles. mAb nCoV396 is illustrated with surface representation. All figures were prepared by Pymol.

    Techniques Used: Binding Assay, Labeling

    Acquisition and characterization of antibodies. Serum antibody titers of six SARS-CoV-2 convalescent patients and a healthy person (ZH0081, non-COVID-19) to the SARS-CoV-2 S ( a ) and N ( b ) proteins measured by ELISA. All samples were performed in triplicates and mean were presented. Sorting of single plasma cells ( c ) with CD38 and CD27 double-positive B cells. d To minimize false positives, each of the S1 and N proteins labeled with Phycoerythrin-canin7 (PE-Cy7) and Brilliant Violet (BV421) was used to sort antigen-specific memory B cells by FACS. e Percentage of different isotypes, VH and VL gene families of 32 isolated N-reactive antibodies. f Number of mutations in nucleotides and amino acids in VH and VL (Vκ and Vλ) of 32 N-reactive antibodies and eight S-reactive antibodies ( g ). Length of the 32 N-reactive antibodies ( h ) and eight S-reactive antibodies ( i ) in H-CDR3. f – i Data are presented as dot and mean values.
    Figure Legend Snippet: Acquisition and characterization of antibodies. Serum antibody titers of six SARS-CoV-2 convalescent patients and a healthy person (ZH0081, non-COVID-19) to the SARS-CoV-2 S ( a ) and N ( b ) proteins measured by ELISA. All samples were performed in triplicates and mean were presented. Sorting of single plasma cells ( c ) with CD38 and CD27 double-positive B cells. d To minimize false positives, each of the S1 and N proteins labeled with Phycoerythrin-canin7 (PE-Cy7) and Brilliant Violet (BV421) was used to sort antigen-specific memory B cells by FACS. e Percentage of different isotypes, VH and VL gene families of 32 isolated N-reactive antibodies. f Number of mutations in nucleotides and amino acids in VH and VL (Vκ and Vλ) of 32 N-reactive antibodies and eight S-reactive antibodies ( g ). Length of the 32 N-reactive antibodies ( h ) and eight S-reactive antibodies ( i ) in H-CDR3. f – i Data are presented as dot and mean values.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Labeling, FACS, Isolation

    3) Product Images from "Original antigenic sin responses to heterologous Betacoronavirus spike proteins are observed in mice following intramuscular administration, but are not apparent in children following SARS-CoV-2 infection"

    Article Title: Original antigenic sin responses to heterologous Betacoronavirus spike proteins are observed in mice following intramuscular administration, but are not apparent in children following SARS-CoV-2 infection

    Journal: medRxiv

    doi: 10.1101/2021.04.29.21256344

    HKU1 antibodies are prevalent in healthy children and children with acute COVID-19 and MIS-C. SARS-CoV-2 (A) and HKU1 (B) spike IgG antibody titers and FRNT neutralization titers (C) in healthy pediatric controls compared to children hospitalized with acute COVID-19 and MIS-C. * P
    Figure Legend Snippet: HKU1 antibodies are prevalent in healthy children and children with acute COVID-19 and MIS-C. SARS-CoV-2 (A) and HKU1 (B) spike IgG antibody titers and FRNT neutralization titers (C) in healthy pediatric controls compared to children hospitalized with acute COVID-19 and MIS-C. * P

    Techniques Used: Neutralization

    Schematic of intramuscular spike protein administrations in groups of five BALB/c mice. Group 1 received prime and boost with SARS-CoV-2 spike, followed by prime and boost with HKU1 spike. Group 2 received a reciprocal administration regimen, with prime and boost with HKU1 spike, followed by prime and boost by HKU1 spike. D, days post-administration; S, spike; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2. *These mice were immunized with nucleocapsid protein 21 and 42 days prior to utilization for this study.
    Figure Legend Snippet: Schematic of intramuscular spike protein administrations in groups of five BALB/c mice. Group 1 received prime and boost with SARS-CoV-2 spike, followed by prime and boost with HKU1 spike. Group 2 received a reciprocal administration regimen, with prime and boost with HKU1 spike, followed by prime and boost by HKU1 spike. D, days post-administration; S, spike; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2. *These mice were immunized with nucleocapsid protein 21 and 42 days prior to utilization for this study.

    Techniques Used: Mouse Assay

    HKU1 spike IgG antibodies correlated positively with both SAR-CoV-2 spike IgG and SARS-CoV-2 neutralizing antibodies in children with acute COVID-19 and MIS-C. Linear regression analyses compared the log-transformed antibody titers of (A) SARS-CoV-2 spike IgG vs. HKU1 spike IgG; (B) HKU1 spike IgG vs. SARS-CoV-2 neutralization titers; and (C) SARS-CoV-2 spike IgG vs. SARS-CoV-2 neutralization titers among children with acute COVID-19 or MIS-C. Spearman’s correlation coefficients (r) and P-values are shown.
    Figure Legend Snippet: HKU1 spike IgG antibodies correlated positively with both SAR-CoV-2 spike IgG and SARS-CoV-2 neutralizing antibodies in children with acute COVID-19 and MIS-C. Linear regression analyses compared the log-transformed antibody titers of (A) SARS-CoV-2 spike IgG vs. HKU1 spike IgG; (B) HKU1 spike IgG vs. SARS-CoV-2 neutralization titers; and (C) SARS-CoV-2 spike IgG vs. SARS-CoV-2 neutralization titers among children with acute COVID-19 or MIS-C. Spearman’s correlation coefficients (r) and P-values are shown.

    Techniques Used: Transformation Assay, Neutralization

    Priming mice with HKU1 spike protein prior to boosting with SARS-CoV-2 spike protein completely impeded the development of SARS-CoV-2 neutralizing antibodies. SARS-CoV-2 (A,B) and HKU1 (C,D) full-length spike IgG binding and SARS-CoV-2 neutralizing (E, F) antibodies in mice are shown as log(end-point titer). Group 1 was primed with two doses of alum-adjuvanted SARS-CoV-2 spike and boosted with two doses of alum-adjuvanted HKU1 spike (A, C, E). Group 2 received the reciprocal regimen of HKU1 spike prime and SARS-CoV-2 spike boost (B, D, F). * P
    Figure Legend Snippet: Priming mice with HKU1 spike protein prior to boosting with SARS-CoV-2 spike protein completely impeded the development of SARS-CoV-2 neutralizing antibodies. SARS-CoV-2 (A,B) and HKU1 (C,D) full-length spike IgG binding and SARS-CoV-2 neutralizing (E, F) antibodies in mice are shown as log(end-point titer). Group 1 was primed with two doses of alum-adjuvanted SARS-CoV-2 spike and boosted with two doses of alum-adjuvanted HKU1 spike (A, C, E). Group 2 received the reciprocal regimen of HKU1 spike prime and SARS-CoV-2 spike boost (B, D, F). * P

    Techniques Used: Mouse Assay, Binding Assay

    4) Product Images from "SARS-CoV-2 RapidPlex: A Graphene-Based Multiplexed Telemedicine Platform for Rapid and Low-Cost COVID-19 Diagnosis and Monitoring"

    Article Title: SARS-CoV-2 RapidPlex: A Graphene-Based Multiplexed Telemedicine Platform for Rapid and Low-Cost COVID-19 Diagnosis and Monitoring

    Journal: Matter

    doi: 10.1016/j.matt.2020.09.027

    Evaluation of Analytical Sensor Performance for the Detection of Physiological Levels of Target COVID-19 Biomarkers (A) Scheme of sensor preparation for detection of SARS-CoV-2 NP and CRP based on double-sandwich and sandwich assay configurations, respectively. CAb, capture antibody; DAb, detector antibody; DAb 2 , secondary detector antibody; HRP, horseradish peroxidase. (B and C) Calibration curves constructed for NP (B) and CRP (C) detection in PBS (pH 7.4) supplemented with 1.0% BSA. Data are presented as mean ± SD (n = 3). (D) Scheme of sensor preparation for detection of S1-IgG and S1-IgM isotypes based on direct assay configurations. (E and F) Calibration curves constructed for S1-IgG (E) and S1-IgM (F) isotype detection in PBS (pH 7.4) supplemented with 1.0% BSA. Data are presented as mean ± SD (n = 3).
    Figure Legend Snippet: Evaluation of Analytical Sensor Performance for the Detection of Physiological Levels of Target COVID-19 Biomarkers (A) Scheme of sensor preparation for detection of SARS-CoV-2 NP and CRP based on double-sandwich and sandwich assay configurations, respectively. CAb, capture antibody; DAb, detector antibody; DAb 2 , secondary detector antibody; HRP, horseradish peroxidase. (B and C) Calibration curves constructed for NP (B) and CRP (C) detection in PBS (pH 7.4) supplemented with 1.0% BSA. Data are presented as mean ± SD (n = 3). (D) Scheme of sensor preparation for detection of S1-IgG and S1-IgM isotypes based on direct assay configurations. (E and F) Calibration curves constructed for S1-IgG (E) and S1-IgM (F) isotype detection in PBS (pH 7.4) supplemented with 1.0% BSA. Data are presented as mean ± SD (n = 3).

    Techniques Used: Construct

    Application of SARS-CoV-2 RapidPlex in SARS-CoV-2 Detection in Blood and Saliva Samples from COVID-19-Positive and -Negative Subjects (A and B) Experimental readings obtained with SARS-CoV-2 RapidPlex after 10-min incubation of the sensor array with serum samples from a representative COVID-19 RT-PCR-negative (A) and -positive (B) patient. (C) Signal of individual sensor obtained after 1-min incubation with a serum sample from a COVID-19-positive patient (dark color) versus the signal obtained after 10-min incubation with a serum sample from a COVID-19-negative patient (light color). (D) Box-and-whisker plot of measured signal-to-blank ratios (S/B) for NP, S1-IgG, S1-IgM, and CRP in RT-PCR-confirmed COVID-19-positive (n = 5) and -negative (n = 6) serum samples. (E) Box-and-whisker plot of measured S/B for NP, S1-IgG, S1-IgM, and CRP in RT-PCR-confirmed COVID-19-positive (n = 5) and -negative (n = 3) saliva samples. (F) CRP levels in diluted serum samples plotted against given COVID-19 symptom severity, with “Healthy” referring to COVID-19-negative patient samples (n = 7). Positive COVID-19 patients were classified according to disease severity as asymptomatic (n = 2), mild (n = 5), and moderate (n = 2).
    Figure Legend Snippet: Application of SARS-CoV-2 RapidPlex in SARS-CoV-2 Detection in Blood and Saliva Samples from COVID-19-Positive and -Negative Subjects (A and B) Experimental readings obtained with SARS-CoV-2 RapidPlex after 10-min incubation of the sensor array with serum samples from a representative COVID-19 RT-PCR-negative (A) and -positive (B) patient. (C) Signal of individual sensor obtained after 1-min incubation with a serum sample from a COVID-19-positive patient (dark color) versus the signal obtained after 10-min incubation with a serum sample from a COVID-19-negative patient (light color). (D) Box-and-whisker plot of measured signal-to-blank ratios (S/B) for NP, S1-IgG, S1-IgM, and CRP in RT-PCR-confirmed COVID-19-positive (n = 5) and -negative (n = 6) serum samples. (E) Box-and-whisker plot of measured S/B for NP, S1-IgG, S1-IgM, and CRP in RT-PCR-confirmed COVID-19-positive (n = 5) and -negative (n = 3) saliva samples. (F) CRP levels in diluted serum samples plotted against given COVID-19 symptom severity, with “Healthy” referring to COVID-19-negative patient samples (n = 7). Positive COVID-19 patients were classified according to disease severity as asymptomatic (n = 2), mild (n = 5), and moderate (n = 2).

    Techniques Used: Incubation, Reverse Transcription Polymerase Chain Reaction, Whisker Assay

    5) Product Images from "COVID-19 Patients Upregulate Toll-like Receptor 4-mediated Inflammatory Signaling That Mimics Bacterial Sepsis"

    Article Title: COVID-19 Patients Upregulate Toll-like Receptor 4-mediated Inflammatory Signaling That Mimics Bacterial Sepsis

    Journal: Journal of Korean Medical Science

    doi: 10.3346/jkms.2020.35.e343

    COVID-19 infection boosts NF-κB signaling pathway. ( A ) The left (MILD) and right (SEVERE) sides of box represent the mean fold change in mRNA levels, compared with HC. The NF-κB signaling pathway was adopted from KEGG database (accession number: hsa04064). ( B ) The expression levels of IRF3 , TLR3 , TLR7 , TLR8 and TLR9 were represented by FPKM. Error bar indicates standard error of mean. P values were calculated using Mann-Whitney U test and adjusted P values (FDR) were shown. COVID-19 = coronavirus disease 2019, NF = nuclear factor, MILD = mild/moderate, SEVERE = severe/critical, HC = healthy controls, KEGG = Kyoto Encyclopedia of Genes and Genomes, FPKM = fragments per kilobase exon-model per million reads mapped, FDR = false discovery rate, TLR = toll-like receptor, IRF = interferon regulatory factor. * P
    Figure Legend Snippet: COVID-19 infection boosts NF-κB signaling pathway. ( A ) The left (MILD) and right (SEVERE) sides of box represent the mean fold change in mRNA levels, compared with HC. The NF-κB signaling pathway was adopted from KEGG database (accession number: hsa04064). ( B ) The expression levels of IRF3 , TLR3 , TLR7 , TLR8 and TLR9 were represented by FPKM. Error bar indicates standard error of mean. P values were calculated using Mann-Whitney U test and adjusted P values (FDR) were shown. COVID-19 = coronavirus disease 2019, NF = nuclear factor, MILD = mild/moderate, SEVERE = severe/critical, HC = healthy controls, KEGG = Kyoto Encyclopedia of Genes and Genomes, FPKM = fragments per kilobase exon-model per million reads mapped, FDR = false discovery rate, TLR = toll-like receptor, IRF = interferon regulatory factor. * P

    Techniques Used: Infection, Expressing, MANN-WHITNEY

    Transcriptome analysis reveals that immune gene expression profiles of COVID-19 patients are distinct to HC. ( A ) Schematic diagram of the immune transcriptome analysis in this study. ( B ) A result of principal component analysis of log2-transformed 579 immune gene expression levels. ( C ) The scatter plots representing 579 immune genes with the log2-transformed FPKM for COVID-19 patients compared to HC. ( D ) The ten most significantly enriched KEGG pathways of the 298 DEiGs from COVID-19 patients compared to HC. ( E ) Log2-transformed fold changes of chemokine and chemokine receptor genes from MILD (x-axis) and SEVERE (y-axis) vs. HC. ( F ) Expression levels (FPKM) of marked chemokines in (E). Error bar indicates standard error of mean. P values were calculated using Mann-Whitney U test and adjusted P values (FDR) were shown. COVID-19 = coronavirus disease 2019, HC = healthy controls, FPKM = fragments per kilobase exon-model per million reads mapped, KEGG = Kyoto Encyclopedia of Genes and Genomes, DEiG = differentially expressed immune gene, MILD = mild/moderate, SEVERE = severe/critical, FDR = false discovery rate, IBD = inflammatory bowel disease, TNF = tumor necrosis factor, CCL = C-C motif chemokine ligand, CXCL = C-X-C motif chemokine ligand. * P
    Figure Legend Snippet: Transcriptome analysis reveals that immune gene expression profiles of COVID-19 patients are distinct to HC. ( A ) Schematic diagram of the immune transcriptome analysis in this study. ( B ) A result of principal component analysis of log2-transformed 579 immune gene expression levels. ( C ) The scatter plots representing 579 immune genes with the log2-transformed FPKM for COVID-19 patients compared to HC. ( D ) The ten most significantly enriched KEGG pathways of the 298 DEiGs from COVID-19 patients compared to HC. ( E ) Log2-transformed fold changes of chemokine and chemokine receptor genes from MILD (x-axis) and SEVERE (y-axis) vs. HC. ( F ) Expression levels (FPKM) of marked chemokines in (E). Error bar indicates standard error of mean. P values were calculated using Mann-Whitney U test and adjusted P values (FDR) were shown. COVID-19 = coronavirus disease 2019, HC = healthy controls, FPKM = fragments per kilobase exon-model per million reads mapped, KEGG = Kyoto Encyclopedia of Genes and Genomes, DEiG = differentially expressed immune gene, MILD = mild/moderate, SEVERE = severe/critical, FDR = false discovery rate, IBD = inflammatory bowel disease, TNF = tumor necrosis factor, CCL = C-C motif chemokine ligand, CXCL = C-X-C motif chemokine ligand. * P

    Techniques Used: Expressing, Transformation Assay, MANN-WHITNEY

    6) Product Images from "The Characterization of Disease Severity Associated IgG Subclasses Response in COVID-19 Patients"

    Article Title: The Characterization of Disease Severity Associated IgG Subclasses Response in COVID-19 Patients

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2021.632814

    IgG1 and IgG3 were the main subclasses induced in COVID-19 patients and related with disease severity. IgG1 (A) and IgG3 (B) responses to NP, S, and RBD and neutralizing antibody (C) against SARS-CoV-2 pseudo-virus with luciferase reporter gene in the healthy controls ( n = 11) and the severe ( n = 24), moderate ( n = 54), mild ( n = 35), asymptomatic ( n = 10) patients were detected. Differences of medium values between groups were analyzed by Mann-Whitney U -test. Significant correlations among Severity, NAb and IgG subclasses (D) including anti-NP IgG1, IgG3, anti-S IgG1, IgG3, anti-RBD IgG1, IgG3 were shown. Spearman correlation coefficient was calculated. A two-tailed P value
    Figure Legend Snippet: IgG1 and IgG3 were the main subclasses induced in COVID-19 patients and related with disease severity. IgG1 (A) and IgG3 (B) responses to NP, S, and RBD and neutralizing antibody (C) against SARS-CoV-2 pseudo-virus with luciferase reporter gene in the healthy controls ( n = 11) and the severe ( n = 24), moderate ( n = 54), mild ( n = 35), asymptomatic ( n = 10) patients were detected. Differences of medium values between groups were analyzed by Mann-Whitney U -test. Significant correlations among Severity, NAb and IgG subclasses (D) including anti-NP IgG1, IgG3, anti-S IgG1, IgG3, anti-RBD IgG1, IgG3 were shown. Spearman correlation coefficient was calculated. A two-tailed P value

    Techniques Used: Luciferase, MANN-WHITNEY, Two Tailed Test

    IgA, IgG, and IgM antibodies responses in COVID-19 ranging from asymptomatic to severe patients. Serum samples collected from COVID-19 patients were used for detecting IgA, IgG, and IgM levels to NP (A) , S (B) , and RBD (C) antigens of SARS-CoV-2 via ELISA. Antibody titers of the healthy controls ( n = 11) and the severe ( n = 24), moderate ( n = 54), mild ( n = 35), asymptomatic ( n = 10) patients were shown in (A–C) . Mann-Whitney U -test was used to compare differences of medium values between groups, a two-tailed P value
    Figure Legend Snippet: IgA, IgG, and IgM antibodies responses in COVID-19 ranging from asymptomatic to severe patients. Serum samples collected from COVID-19 patients were used for detecting IgA, IgG, and IgM levels to NP (A) , S (B) , and RBD (C) antigens of SARS-CoV-2 via ELISA. Antibody titers of the healthy controls ( n = 11) and the severe ( n = 24), moderate ( n = 54), mild ( n = 35), asymptomatic ( n = 10) patients were shown in (A–C) . Mann-Whitney U -test was used to compare differences of medium values between groups, a two-tailed P value

    Techniques Used: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Two Tailed Test

    7) Product Images from "SARS-CoV-2–Specific Antibody Detection for Seroepidemiology: A Multiplex Analysis Approach Accounting for Accurate Seroprevalence"

    Article Title: SARS-CoV-2–Specific Antibody Detection for Seroepidemiology: A Multiplex Analysis Approach Accounting for Accurate Seroprevalence

    Journal: The Journal of Infectious Diseases

    doi: 10.1093/infdis/jiaa479

    Discrimination of COVID-19 patients with varying severity from a cross-sectional population panel and ILI patients. A , Individuals from the cross-sectional panel aged 3–90 years (n = 224), ILI patients with noncoronavirus (n = 75), and non-SARS-CoV-2 seasonal coronavirus-infected ILI patients (n = 109) were compared to hospitalized and nonhospitalized COVID-19 patients. Median concentration and 95% confidence intervals and statistical results (adjusted P values of Tukey multiple comparison) between the groups are shown. B , Laboratory-confirmed viral infections (see Supplementary Table 2 ) and concentration data of ILI patients are shown to confirm that the assay discriminates SARS-CoV-2–specific antibodies from antibodies induced by various laboratory-confirmed viral infections. Abbreviations: AU, arbitrary unit; COVID-19, coronavirus disease 2019; HCoV, human coronavirus; MERS-CoV, Middle East respiratory syndrome coronavirus; N, nucleoprotein; non-HCoV, noncoronavirus; RBD, receptor binding domain; RSV, respiratory syncytial virus; S1, spike protein subunit 1.
    Figure Legend Snippet: Discrimination of COVID-19 patients with varying severity from a cross-sectional population panel and ILI patients. A , Individuals from the cross-sectional panel aged 3–90 years (n = 224), ILI patients with noncoronavirus (n = 75), and non-SARS-CoV-2 seasonal coronavirus-infected ILI patients (n = 109) were compared to hospitalized and nonhospitalized COVID-19 patients. Median concentration and 95% confidence intervals and statistical results (adjusted P values of Tukey multiple comparison) between the groups are shown. B , Laboratory-confirmed viral infections (see Supplementary Table 2 ) and concentration data of ILI patients are shown to confirm that the assay discriminates SARS-CoV-2–specific antibodies from antibodies induced by various laboratory-confirmed viral infections. Abbreviations: AU, arbitrary unit; COVID-19, coronavirus disease 2019; HCoV, human coronavirus; MERS-CoV, Middle East respiratory syndrome coronavirus; N, nucleoprotein; non-HCoV, noncoronavirus; RBD, receptor binding domain; RSV, respiratory syncytial virus; S1, spike protein subunit 1.

    Techniques Used: Infection, Concentration Assay, Binding Assay

    Kinetics of antibody production after disease onset in hospitalized and nonhospitalized COVID-19 patients. Paired samples were analyzed to identify changes in IgG concentrations in hospitalized ( A ) and nonhospitalized ( B ) COVID-19 patients. C , The log-transformed concentration data of the samples shown in ( A ) and ( B ) were fitted with a 4-parameter nonlinear least squared fit. D , Of each patient with paired samples available, 1 sample was selected randomly and data were fitted to estimate the slope, R 2 of the fits, and the difference between the fitted lines determined. Abbreviations: AU, arbitrary unit; COVID-19, coronavirus disease 2019; IgG, immunoglobulin G; N, nucleoprotein; RBD, receptor binding domain; S1, spike protein subunit 1.
    Figure Legend Snippet: Kinetics of antibody production after disease onset in hospitalized and nonhospitalized COVID-19 patients. Paired samples were analyzed to identify changes in IgG concentrations in hospitalized ( A ) and nonhospitalized ( B ) COVID-19 patients. C , The log-transformed concentration data of the samples shown in ( A ) and ( B ) were fitted with a 4-parameter nonlinear least squared fit. D , Of each patient with paired samples available, 1 sample was selected randomly and data were fitted to estimate the slope, R 2 of the fits, and the difference between the fitted lines determined. Abbreviations: AU, arbitrary unit; COVID-19, coronavirus disease 2019; IgG, immunoglobulin G; N, nucleoprotein; RBD, receptor binding domain; S1, spike protein subunit 1.

    Techniques Used: Transformation Assay, Concentration Assay, Binding Assay

    8) Product Images from "SARS-CoV-2 infection induces germinal center responses with robust stimulation of CD4 T follicular helper cells in rhesus macaques"

    Article Title: SARS-CoV-2 infection induces germinal center responses with robust stimulation of CD4 T follicular helper cells in rhesus macaques

    Journal: bioRxiv

    doi: 10.1101/2020.07.07.191007

    Humoral responses to SARS-CoV-2 are dominated by IgG antibodies Concentrations of (A) IgM, (B) IgG, and (C) IgA antibodies specific for S1, S2, and N proteins were measured by BAMA or ELISA in serum of macaques infused with human COVID-19 convalescent plasma (CP; blue symbols) or naive plasma (NP; red symbols) and control non-infused animals (black symbols). The dashed line represents the median pre-infection (day 0) concentration for all animals. (D) The magnitude of the IgM, IgG and IgA antibody responses in animals that were not given human convalescent plasma was determined by dividing post-infection concentrations by those measured on day 0 in each animal. Geometric mean fold increases with SEM are shown. (E) Correlations between day 10 levels of S1-specific IgG and IgM, N-specific IgA and IgG, and pseudovirus neutralizing antibody titers and anti-RBD IgG antibodies measured by ELISA.
    Figure Legend Snippet: Humoral responses to SARS-CoV-2 are dominated by IgG antibodies Concentrations of (A) IgM, (B) IgG, and (C) IgA antibodies specific for S1, S2, and N proteins were measured by BAMA or ELISA in serum of macaques infused with human COVID-19 convalescent plasma (CP; blue symbols) or naive plasma (NP; red symbols) and control non-infused animals (black symbols). The dashed line represents the median pre-infection (day 0) concentration for all animals. (D) The magnitude of the IgM, IgG and IgA antibody responses in animals that were not given human convalescent plasma was determined by dividing post-infection concentrations by those measured on day 0 in each animal. Geometric mean fold increases with SEM are shown. (E) Correlations between day 10 levels of S1-specific IgG and IgM, N-specific IgA and IgG, and pseudovirus neutralizing antibody titers and anti-RBD IgG antibodies measured by ELISA.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Infection, Concentration Assay

    SARS-CoV-2 infection leads to a rapid and transient shift in innate immune responses and increases the number CD4 T follicular helper cells in peripheral blood. (A) Experimental design. Indian-origin rhesus macaques were inoculated with SARS-CoV-2 (SARS-CoV-2/human/USA/CA-CZB-59×002/2020) via the intranasal (IN), intratracheal (IT) and ocular route. Twenty-four hours later, animals were infused with either COVID-19 convalescent human plasma (I+CP; blue symbols), or normal plasma (I+NP; red symbols) (both at 4ml/kg), and four animals did not receive any plasma (infected; black symbols). Blood was sampled over the course of infection and tissues were collected at necropsy (11-14 DPI) for immune profiling. (B) Mean viral RNA (+range) in each of the groups within nasal washes (C) Flow plot illustrating gating strategy to identify innate immune subsets in whole blood. (D ) Kinetics of innate immune responses (*p
    Figure Legend Snippet: SARS-CoV-2 infection leads to a rapid and transient shift in innate immune responses and increases the number CD4 T follicular helper cells in peripheral blood. (A) Experimental design. Indian-origin rhesus macaques were inoculated with SARS-CoV-2 (SARS-CoV-2/human/USA/CA-CZB-59×002/2020) via the intranasal (IN), intratracheal (IT) and ocular route. Twenty-four hours later, animals were infused with either COVID-19 convalescent human plasma (I+CP; blue symbols), or normal plasma (I+NP; red symbols) (both at 4ml/kg), and four animals did not receive any plasma (infected; black symbols). Blood was sampled over the course of infection and tissues were collected at necropsy (11-14 DPI) for immune profiling. (B) Mean viral RNA (+range) in each of the groups within nasal washes (C) Flow plot illustrating gating strategy to identify innate immune subsets in whole blood. (D ) Kinetics of innate immune responses (*p

    Techniques Used: Infection

    9) Product Images from "Evaluation of Humoral Immunity to SARS-CoV-2: Diagnostic Value of a New Multiplex Addressable Laser Bead Immunoassay"

    Article Title: Evaluation of Humoral Immunity to SARS-CoV-2: Diagnostic Value of a New Multiplex Addressable Laser Bead Immunoassay

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2020.603931

    Detection, titration, and cross-reactivity of anti–SARS-CoV-2 Spike S1, nucleocapsid N protein IgG, and anti–SARS-CoV-2 Spike S1 IgM antibodies by ALBIA-IgG-S1/N and ALBIA-IgM-S1. (A) A calibration curve was obtained after serial dilutions of the calibrator, i.e., one highly positive sample. A plateau of MFI was reached for dilutions 1:400 or lower. (B) Calculation of antibody titer by reference to the MFI value of the calibrator (gray bar) used at a 1:400 dilution in the assay and its level arbitrarily set to 100 arbitrary units (AU)/mL. The assay was first performed using a 1:100 screening dilution of the serum. In case the sample’s MFI at 1/100 dilution was higher than 70% of the calibrator’s MFI, further dilutions were performed, and the first dilution yielding an MFI inferior to 70% of calibrator MFI was retained for calculation. An example is given: at 1:100 dilution, the MFI was higher than 70% of the calibrator’s MFI (23,311 × 0.7 = 16,318), requiring a 1/800 dilution for computing the titer, i.e., 94 AU/mL anti-S1 IgG level. Specificity toward non–COVID-19 patients: (C) anti-Spike S1 and (D) anti-N IgG, IgM, and (E) anti-Spike S1 IgM antibody reactivity in patients with different conditions: PCR-confirmed infection with other CoV (17 sera from 13 patients; HKU1, n = 3; OC43, n = 11; NL63, n = 3). RA, rheumatoid arthritis; SS, Sjögren syndrome; ASS, antisynthetase syndrome; SLE, systemic lupus erythematosus.
    Figure Legend Snippet: Detection, titration, and cross-reactivity of anti–SARS-CoV-2 Spike S1, nucleocapsid N protein IgG, and anti–SARS-CoV-2 Spike S1 IgM antibodies by ALBIA-IgG-S1/N and ALBIA-IgM-S1. (A) A calibration curve was obtained after serial dilutions of the calibrator, i.e., one highly positive sample. A plateau of MFI was reached for dilutions 1:400 or lower. (B) Calculation of antibody titer by reference to the MFI value of the calibrator (gray bar) used at a 1:400 dilution in the assay and its level arbitrarily set to 100 arbitrary units (AU)/mL. The assay was first performed using a 1:100 screening dilution of the serum. In case the sample’s MFI at 1/100 dilution was higher than 70% of the calibrator’s MFI, further dilutions were performed, and the first dilution yielding an MFI inferior to 70% of calibrator MFI was retained for calculation. An example is given: at 1:100 dilution, the MFI was higher than 70% of the calibrator’s MFI (23,311 × 0.7 = 16,318), requiring a 1/800 dilution for computing the titer, i.e., 94 AU/mL anti-S1 IgG level. Specificity toward non–COVID-19 patients: (C) anti-Spike S1 and (D) anti-N IgG, IgM, and (E) anti-Spike S1 IgM antibody reactivity in patients with different conditions: PCR-confirmed infection with other CoV (17 sera from 13 patients; HKU1, n = 3; OC43, n = 11; NL63, n = 3). RA, rheumatoid arthritis; SS, Sjögren syndrome; ASS, antisynthetase syndrome; SLE, systemic lupus erythematosus.

    Techniques Used: Titration, Polymerase Chain Reaction, Infection

    10) Product Images from "SARS-CoV-2 Proteome Microarray for Mapping COVID-19 Antibody Interactions at Amino Acid Resolution"

    Article Title: SARS-CoV-2 Proteome Microarray for Mapping COVID-19 Antibody Interactions at Amino Acid Resolution

    Journal: ACS Central Science

    doi: 10.1021/acscentsci.0c00742

    Landscape of the humoral antibody response to SARS-CoV-2 proteins other than Orf1ab. (a, b) The distribution of human IgM and IgG antibodies to SARS-CoV-2 individual proteins (S, E, M, N, Orf3a, Orf6, Orf7a, Orf8, and Orf10), respectively. The x -axis represents the sequence of amino acids of SARS-CoV-2 proteins from the N-terminal to C-terminal. The y -axis represents the serum samples from COVID-19 patients. The false-colored rainbow color from blue to red corresponds to the signals of antibody binding from low to high, respectively.
    Figure Legend Snippet: Landscape of the humoral antibody response to SARS-CoV-2 proteins other than Orf1ab. (a, b) The distribution of human IgM and IgG antibodies to SARS-CoV-2 individual proteins (S, E, M, N, Orf3a, Orf6, Orf7a, Orf8, and Orf10), respectively. The x -axis represents the sequence of amino acids of SARS-CoV-2 proteins from the N-terminal to C-terminal. The y -axis represents the serum samples from COVID-19 patients. The false-colored rainbow color from blue to red corresponds to the signals of antibody binding from low to high, respectively.

    Techniques Used: Sequencing, Binding Assay

    Landscape of humoral IgM antibody response to SARS-CoV-2 Orf1ab proteome. The x -axis represents the sequence of amino acids of SARS-CoV-2 nonstructural proteins (nsps) from the N-terminal to C-terminal. The y -axis represents the serum samples from COVID-19 patients. The false-colored rainbow color from blue to red corresponds to the signals of antibody binding from low to high, respectively.
    Figure Legend Snippet: Landscape of humoral IgM antibody response to SARS-CoV-2 Orf1ab proteome. The x -axis represents the sequence of amino acids of SARS-CoV-2 nonstructural proteins (nsps) from the N-terminal to C-terminal. The y -axis represents the serum samples from COVID-19 patients. The false-colored rainbow color from blue to red corresponds to the signals of antibody binding from low to high, respectively.

    Techniques Used: Sequencing, Binding Assay

    Landscape of the humoral IgG antibody response to the SARS-CoV-2 Orf1ab proteome. The x -axis represents the sequence of amino acids of the SARS-CoV-2 nonstructural proteins (nsps) from the N-terminal to C-terminal. The y -axis represents the serum samples from the COVID-19 patients. The false-colored rainbow color from blue to red corresponds to the signals of antibody binding from low to high, respectively.
    Figure Legend Snippet: Landscape of the humoral IgG antibody response to the SARS-CoV-2 Orf1ab proteome. The x -axis represents the sequence of amino acids of the SARS-CoV-2 nonstructural proteins (nsps) from the N-terminal to C-terminal. The y -axis represents the serum samples from the COVID-19 patients. The false-colored rainbow color from blue to red corresponds to the signals of antibody binding from low to high, respectively.

    Techniques Used: Sequencing, Binding Assay

    Identification of potential peptide epitopes for SARS-CoV-2 detection and neutralization. (a) Box-plot analysis of antibody responses to immunogenic epitopes of SARS-COV-2 between COVID-19 patients and control patients. The significance was performed using the Mann–Whitney U-test ( p -value
    Figure Legend Snippet: Identification of potential peptide epitopes for SARS-CoV-2 detection and neutralization. (a) Box-plot analysis of antibody responses to immunogenic epitopes of SARS-COV-2 between COVID-19 patients and control patients. The significance was performed using the Mann–Whitney U-test ( p -value

    Techniques Used: Neutralization, MANN-WHITNEY

    11) Product Images from "Clonal dissection of immunodominance and cross-reactivity of the CD4+ T cell response to SARS-CoV-2"

    Article Title: Clonal dissection of immunodominance and cross-reactivity of the CD4+ T cell response to SARS-CoV-2

    Journal: bioRxiv

    doi: 10.1101/2021.03.23.436642

    Sorting of T cell subsets and identification of SARS-CoV-2 Spike- and Nucleoprotein-reactive CD4 + T cells in COVID-19 and pre-pandemic samples. ( A ) Sorting strategy to isolate CD4 + total memory T cells and Tcm, Tem and cTfh subsets. ( B, C ) Characterization of antigen-specific T cells by CFSE dilution combined with CD25 and ICOS co-expression at day 7 following stimulation with Spike or Nucleoprotein in the presence of autologous monocytes. Negative controls of T cells cultured with monocytes alone are reported as dashed lines. Shown are data from patient P2 and from a pre-pandemic healthy donor sample (HD1).
    Figure Legend Snippet: Sorting of T cell subsets and identification of SARS-CoV-2 Spike- and Nucleoprotein-reactive CD4 + T cells in COVID-19 and pre-pandemic samples. ( A ) Sorting strategy to isolate CD4 + total memory T cells and Tcm, Tem and cTfh subsets. ( B, C ) Characterization of antigen-specific T cells by CFSE dilution combined with CD25 and ICOS co-expression at day 7 following stimulation with Spike or Nucleoprotein in the presence of autologous monocytes. Negative controls of T cells cultured with monocytes alone are reported as dashed lines. Shown are data from patient P2 and from a pre-pandemic healthy donor sample (HD1).

    Techniques Used: Transmission Electron Microscopy, Expressing, Cell Culture

    12) Product Images from "Nucleocapsid and Spike Proteins of SARS-CoV-2 Drive Neutrophil Extracellular Trap Formation"

    Article Title: Nucleocapsid and Spike Proteins of SARS-CoV-2 Drive Neutrophil Extracellular Trap Formation

    Journal: Immune Network

    doi: 10.4110/in.2021.21.e16

    N and S proteins of SARS-CoV-2 induce NET formation from neutrophils. (A-C) Neutrophils were incubated with various concentrations (1, 10, and 100 nM) of the N protein, whole S protein, S1 subunits of the S protein, or S2 subunits of the S protein for 2 h. (A) NET formation in response to viral proteins was determined by Sytox Green staining. (B) Representative immunofluorescence images of NET formation in response to viral proteins of SARS-CoV-2. Representative images of more than 5 experiments are shown (scale bar, 10 μm). (C) ROS generation in response to viral proteins was determined by DCF-DA staining. (D, E) The effects of viral proteins on chemotaxis of neutrophils. One side of chamber was coated with either N, S, S1, or S2 protein and chemotaxis of neutrophils toward viral proteins was examined. (D) Neutrophil migration tracking analysis. The distances traveled by neutrophils were tracked for 45 min. Representative tracking results of thirty cells per each group are shown (n=3 per group). (E) Relative mean distance and relative mean velocity of neutrophils migrating toward viral proteins. Data are expressed as means±SEMs. Con, control; MPO, myeloperoxidase; H3Cit, citrullinated histone 3. * p
    Figure Legend Snippet: N and S proteins of SARS-CoV-2 induce NET formation from neutrophils. (A-C) Neutrophils were incubated with various concentrations (1, 10, and 100 nM) of the N protein, whole S protein, S1 subunits of the S protein, or S2 subunits of the S protein for 2 h. (A) NET formation in response to viral proteins was determined by Sytox Green staining. (B) Representative immunofluorescence images of NET formation in response to viral proteins of SARS-CoV-2. Representative images of more than 5 experiments are shown (scale bar, 10 μm). (C) ROS generation in response to viral proteins was determined by DCF-DA staining. (D, E) The effects of viral proteins on chemotaxis of neutrophils. One side of chamber was coated with either N, S, S1, or S2 protein and chemotaxis of neutrophils toward viral proteins was examined. (D) Neutrophil migration tracking analysis. The distances traveled by neutrophils were tracked for 45 min. Representative tracking results of thirty cells per each group are shown (n=3 per group). (E) Relative mean distance and relative mean velocity of neutrophils migrating toward viral proteins. Data are expressed as means±SEMs. Con, control; MPO, myeloperoxidase; H3Cit, citrullinated histone 3. * p

    Techniques Used: Incubation, Staining, Immunofluorescence, Chemotaxis Assay, Migration

    13) Product Images from "Novel ELISA Protocol Links Pre-Existing SARS-CoV-2 Reactive Antibodies With Endemic Coronavirus Immunity and Age and Reveals Improved Serologic Identification of Acute COVID-19 via Multi-Parameter Detection"

    Article Title: Novel ELISA Protocol Links Pre-Existing SARS-CoV-2 Reactive Antibodies With Endemic Coronavirus Immunity and Age and Reveals Improved Serologic Identification of Acute COVID-19 via Multi-Parameter Detection

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2021.614676

    Detection and quantification of SARS-CoV-2 RBD- and N- reactive antibodies in pre-pandemic samples. (A) Representative dilution curves of seven pre-pandemic samples with SARS-CoV-2 RBD-reactive antibodies (three subjects per graph). Open and solid symbols represent buffer only coat and SARS-CoV-2 RBD coat, respectively. Arbitrary Units (AU) were calculated as described in Methods and shown beneath the respective isotype graph for diluent only and SARS-CoV-2 RBD coat. AUs for SARS-CoV-2 RBD (B) and N (C) reactive IgM, IgG, and IgA in pre-pandemic samples. Open and solid symbols represent negative and positive results, respectively, as determined by Metric 1. Enumeration of the positive samples for each isotype in the pre-pandemic cohort is shown beneath each graph with percentages of total in parentheses. (D) Correlation between AUs for IgG reactive to SARS-CoV-2 RBD and N (n=32). Values were log-transformed to obtain a parametric distribution. Statistical analyses were performed using an unpaired non-parametric Mann-Whitney t-test in (B, C) and Pearson’s correlation of normally distributed AU values for (D) .
    Figure Legend Snippet: Detection and quantification of SARS-CoV-2 RBD- and N- reactive antibodies in pre-pandemic samples. (A) Representative dilution curves of seven pre-pandemic samples with SARS-CoV-2 RBD-reactive antibodies (three subjects per graph). Open and solid symbols represent buffer only coat and SARS-CoV-2 RBD coat, respectively. Arbitrary Units (AU) were calculated as described in Methods and shown beneath the respective isotype graph for diluent only and SARS-CoV-2 RBD coat. AUs for SARS-CoV-2 RBD (B) and N (C) reactive IgM, IgG, and IgA in pre-pandemic samples. Open and solid symbols represent negative and positive results, respectively, as determined by Metric 1. Enumeration of the positive samples for each isotype in the pre-pandemic cohort is shown beneath each graph with percentages of total in parentheses. (D) Correlation between AUs for IgG reactive to SARS-CoV-2 RBD and N (n=32). Values were log-transformed to obtain a parametric distribution. Statistical analyses were performed using an unpaired non-parametric Mann-Whitney t-test in (B, C) and Pearson’s correlation of normally distributed AU values for (D) .

    Techniques Used: Transformation Assay, MANN-WHITNEY

    The modified ELISA (BU ELISA) protocol exhibits low background signal at high sample concentration and use of SARS-Cov-2 RBD-recombinant antibody standard curves allows for accurate sample quantification via accounting for OD drift between experimental runs. (A) Dilution curves of buffer only coated wells from five donor samples after using an automated plate washer or the BU ELISA method of multichannel plate washing. Experiment was performed once. (B) Representative dilution curves of buffer only coated wells from 30 subjects, average and range of 1:5 sample dilution for each isotype from all subjects; IgM, IgG, and IgA were detected in individual assays. (C) Representative IgM, IgG, and IgA standard curves from 15 different experimental runs are shown. The average of all runs shown as red triangles.
    Figure Legend Snippet: The modified ELISA (BU ELISA) protocol exhibits low background signal at high sample concentration and use of SARS-Cov-2 RBD-recombinant antibody standard curves allows for accurate sample quantification via accounting for OD drift between experimental runs. (A) Dilution curves of buffer only coated wells from five donor samples after using an automated plate washer or the BU ELISA method of multichannel plate washing. Experiment was performed once. (B) Representative dilution curves of buffer only coated wells from 30 subjects, average and range of 1:5 sample dilution for each isotype from all subjects; IgM, IgG, and IgA were detected in individual assays. (C) Representative IgM, IgG, and IgA standard curves from 15 different experimental runs are shown. The average of all runs shown as red triangles.

    Techniques Used: Modification, Enzyme-linked Immunosorbent Assay, Concentration Assay, Recombinant

    SARS-CoV-2 RBD and N reactive IgG in pre-pandemic samples track with IgG recognizing analogous proteins of eCoV strains. (A) AUs of IgG reactive to RBD of NL63 and HKU1 and N of all four eCoV strains (NL63, 2293, OC43, and HKU1). (B) Correlation between SARS-CoV-2 RBD IgG levels with NL63, HKU1 RBD IgG levels in individual subjects. (C) Correlation between SARS-CoV-2 N IgG and NL63, 229E, OC43, and HKU1 N IgG levels, n=30-42. Values were log-transformed to obtain a parametric distribution. Statistical analyses were performed using Pearson’s correlation of normally distributed log transformed AU values in (B, C) and an unpaired non-parametric Mann-Whitney t-test in (A) .
    Figure Legend Snippet: SARS-CoV-2 RBD and N reactive IgG in pre-pandemic samples track with IgG recognizing analogous proteins of eCoV strains. (A) AUs of IgG reactive to RBD of NL63 and HKU1 and N of all four eCoV strains (NL63, 2293, OC43, and HKU1). (B) Correlation between SARS-CoV-2 RBD IgG levels with NL63, HKU1 RBD IgG levels in individual subjects. (C) Correlation between SARS-CoV-2 N IgG and NL63, 229E, OC43, and HKU1 N IgG levels, n=30-42. Values were log-transformed to obtain a parametric distribution. Statistical analyses were performed using Pearson’s correlation of normally distributed log transformed AU values in (B, C) and an unpaired non-parametric Mann-Whitney t-test in (A) .

    Techniques Used: Transformation Assay, MANN-WHITNEY

    Older age is associated with lower circulating antibodies reactive with SARS-CoV-2 and eCoV RBD and N antigens. Quantification of IgG reactive to RBD of NL63, HKU1, and SARS-CoV-2 and N of NL63, 229E, OC43, HKU1, and CoV-2 in pre-pandemic samples regrouped based on HIV (A) or SLE (B) disease status or age (C) . Statistical analyses were performed using an unpaired non-parametric Mann-Whitney t-test.
    Figure Legend Snippet: Older age is associated with lower circulating antibodies reactive with SARS-CoV-2 and eCoV RBD and N antigens. Quantification of IgG reactive to RBD of NL63, HKU1, and SARS-CoV-2 and N of NL63, 229E, OC43, HKU1, and CoV-2 in pre-pandemic samples regrouped based on HIV (A) or SLE (B) disease status or age (C) . Statistical analyses were performed using an unpaired non-parametric Mann-Whitney t-test.

    Techniques Used: MANN-WHITNEY

    Quantification of the relative levels of IgM, IgG, and IgA-reactive SARS-CoV-2-RBD and N antibodies from acute and convalescent SARS-CoV-2 infected subjects. (A) Arbitrary Units (AUs) of SARS-CoV-2 RBD and N reactive IgM, IgG, and IgA of acute and convalescent subjects. Open and solid symbols represent negative and positive results, respectively, as determined by our Metric 1 described in Methods. (B) Correlation between SARS-CoV-2 RBD and N IgM, IgG, and IgA log transformed AUs. Values were log-transformed to obtain a parametric distribution. (C) Quantification of SARS-CoV-2 RBD and N reactive IgM, IgG, and IgA of acute subjects regrouped based on results from Abbott’s SARS-CoV-2 IgG CMIA. Correlation between SARS-CoV-2 RBD (D) and N (E) IgM, IgG, and IgA AUs (log transformed) with the number of days post symptom (dps) onset at time of sample collection for acute subjects. Quantification of SARS-CoV-2 RBD reactive IgM and N reactive IgA (F) and RBD N reactive for IgM, IgG, and IgA (G) for pre-pandemics (n = 19) and Acutes re-classified based on Abbott test results. Light blue bars depict AU range of pre-pandemics for each respective antigen and isotype. Statistical analyses were performed using an unpaired non-parametric Mann-Whitney t-test in (A, C, F, G) and Pearson’s correlation of normally distributed log transformed AU values in (B, D, E) dps, days post symptom.
    Figure Legend Snippet: Quantification of the relative levels of IgM, IgG, and IgA-reactive SARS-CoV-2-RBD and N antibodies from acute and convalescent SARS-CoV-2 infected subjects. (A) Arbitrary Units (AUs) of SARS-CoV-2 RBD and N reactive IgM, IgG, and IgA of acute and convalescent subjects. Open and solid symbols represent negative and positive results, respectively, as determined by our Metric 1 described in Methods. (B) Correlation between SARS-CoV-2 RBD and N IgM, IgG, and IgA log transformed AUs. Values were log-transformed to obtain a parametric distribution. (C) Quantification of SARS-CoV-2 RBD and N reactive IgM, IgG, and IgA of acute subjects regrouped based on results from Abbott’s SARS-CoV-2 IgG CMIA. Correlation between SARS-CoV-2 RBD (D) and N (E) IgM, IgG, and IgA AUs (log transformed) with the number of days post symptom (dps) onset at time of sample collection for acute subjects. Quantification of SARS-CoV-2 RBD reactive IgM and N reactive IgA (F) and RBD N reactive for IgM, IgG, and IgA (G) for pre-pandemics (n = 19) and Acutes re-classified based on Abbott test results. Light blue bars depict AU range of pre-pandemics for each respective antigen and isotype. Statistical analyses were performed using an unpaired non-parametric Mann-Whitney t-test in (A, C, F, G) and Pearson’s correlation of normally distributed log transformed AU values in (B, D, E) dps, days post symptom.

    Techniques Used: Infection, Transformation Assay, MANN-WHITNEY

    14) Product Images from "Development of an ultrasensitive fluorescent immunochromatographic assay based on multilayer quantum dot nanobead for simultaneous detection of SARS-CoV-2 antigen and influenza A virus"

    Article Title: Development of an ultrasensitive fluorescent immunochromatographic assay based on multilayer quantum dot nanobead for simultaneous detection of SARS-CoV-2 antigen and influenza A virus

    Journal: Sensors and Actuators. B, Chemical

    doi: 10.1016/j.snb.2021.130372

    (a) Fluorescence pictures and (b) corresponding test line intensities of the SiQD-ICA, SiDQD-ICA and SiTQD-ICA strips for SARS-CoV-2 NP antigen detection. (c) Reproducibility of the SiTQD-ICA for H1N1 and SARS-CoV-2 NP.
    Figure Legend Snippet: (a) Fluorescence pictures and (b) corresponding test line intensities of the SiQD-ICA, SiDQD-ICA and SiTQD-ICA strips for SARS-CoV-2 NP antigen detection. (c) Reproducibility of the SiTQD-ICA for H1N1 and SARS-CoV-2 NP.

    Techniques Used: Fluorescence

    (a) Photograph, (b) corresponding scanning waveforms of fluorescence, (c) fluorescence intensity of SiTQD-based ICA with different concentration of SARS-CoV-2 NP antigen and FluA H1N1: (i) 10 ng/mL, 10 4 pfu/mL; (ii) 10 ng/mL, 0 pfu/mL; (iii) 0 ng/mL, 10 4 pfu/mL; and (iv) 0 ng/mL, 0 pfu/mL. (d) Typical SEM images of the test zone for SARS-CoV-2 NP antigen concentrations of 10 ng/mL (*) and 0 ng/mL (**). Optimization of (e) SARS-CoV-2 NP capture antibody and (f) FluA capture antibody concentration on the T line. The error bars showed standard deviations calculated from three tests.
    Figure Legend Snippet: (a) Photograph, (b) corresponding scanning waveforms of fluorescence, (c) fluorescence intensity of SiTQD-based ICA with different concentration of SARS-CoV-2 NP antigen and FluA H1N1: (i) 10 ng/mL, 10 4 pfu/mL; (ii) 10 ng/mL, 0 pfu/mL; (iii) 0 ng/mL, 10 4 pfu/mL; and (iv) 0 ng/mL, 0 pfu/mL. (d) Typical SEM images of the test zone for SARS-CoV-2 NP antigen concentrations of 10 ng/mL (*) and 0 ng/mL (**). Optimization of (e) SARS-CoV-2 NP capture antibody and (f) FluA capture antibody concentration on the T line. The error bars showed standard deviations calculated from three tests.

    Techniques Used: Fluorescence, Concentration Assay

    (a) Fluorescence pictures (i) and corresponding test line intensities (ii) of SiTQD-based ICA strip for SARS-CoV-2 NP antigen and H1N1 detection. Corresponding calibration curves for (b) SARS-CoV-2 NP antigen and (c) H1N1. The error bars represented standard deviations calculated from three experiments. (d) Photographs of colloidal gold-based ICA strips for different concentrations of (i) H1N1 and (ii) SARS-CoV-2 NP antigen detection. (e-f) ELISA analysis for SARS-CoV-2 NP antigen (e) and H1N1 detection (f). The insets are colorimetric results of ELISA plates for different concentrations of target virus antigens.
    Figure Legend Snippet: (a) Fluorescence pictures (i) and corresponding test line intensities (ii) of SiTQD-based ICA strip for SARS-CoV-2 NP antigen and H1N1 detection. Corresponding calibration curves for (b) SARS-CoV-2 NP antigen and (c) H1N1. The error bars represented standard deviations calculated from three experiments. (d) Photographs of colloidal gold-based ICA strips for different concentrations of (i) H1N1 and (ii) SARS-CoV-2 NP antigen detection. (e-f) ELISA analysis for SARS-CoV-2 NP antigen (e) and H1N1 detection (f). The insets are colorimetric results of ELISA plates for different concentrations of target virus antigens.

    Techniques Used: Fluorescence, Stripping Membranes, Enzyme-linked Immunosorbent Assay

    (a) Specificity of SiTQD-based ICA. (b) Fluorescence pictures and (c) corresponding test line intensities of SiTQD-based ICA for inactivated SARS-CoV-2 samples. Error bars are calculated from three experiments.
    Figure Legend Snippet: (a) Specificity of SiTQD-based ICA. (b) Fluorescence pictures and (c) corresponding test line intensities of SiTQD-based ICA for inactivated SARS-CoV-2 samples. Error bars are calculated from three experiments.

    Techniques Used: Fluorescence

    Fabrication of SiTQD probes and their application to ICA-based biosensor for simultaneous detection of SARS-CoV-2 and FluA. (a) Synthesis of SiTQD nanocomposite, (b) preparation of immuno-SiTQD probes, and (c) operating principle of SiTQD-ICA strip for detecting two target respiratory viruses.
    Figure Legend Snippet: Fabrication of SiTQD probes and their application to ICA-based biosensor for simultaneous detection of SARS-CoV-2 and FluA. (a) Synthesis of SiTQD nanocomposite, (b) preparation of immuno-SiTQD probes, and (c) operating principle of SiTQD-ICA strip for detecting two target respiratory viruses.

    Techniques Used: Stripping Membranes

    15) Product Images from "A Next Generation Bivalent Human Ad5 COVID-19 Vaccine Delivering Both Spike and Nucleocapsid Antigens Elicits Th1 Dominant CD4+, CD8+ T-cell and Neutralizing Antibody Responses"

    Article Title: A Next Generation Bivalent Human Ad5 COVID-19 Vaccine Delivering Both Spike and Nucleocapsid Antigens Elicits Th1 Dominant CD4+, CD8+ T-cell and Neutralizing Antibody Responses

    Journal: bioRxiv

    doi: 10.1101/2020.07.29.227595

    cPass and Vero E6 cell SARS-CoV-2 confirm neutralization by antibodies. (a) In the cPass assay, inhibition of S RBD interaction with ACE2 was significant at both 1:20 and 1:60 dilutions of serum from hAd5 S-Fusion + N-ETSD vaccinated mice. (b) The results in the Vero E6 cell SARS-CoV-2 viral infection for mice that showed S-specific antibodies by ELISA also showed high neutralization for mice and very high neutralization for pooled sera (G4 pool, blue line) even compared to COVID-19 convalescent serum. G4 pool – mice with S-specific antibodies; M1, M2, M3, M4 – mouse ID; +C – convalescent serum; and media – media only negative control.
    Figure Legend Snippet: cPass and Vero E6 cell SARS-CoV-2 confirm neutralization by antibodies. (a) In the cPass assay, inhibition of S RBD interaction with ACE2 was significant at both 1:20 and 1:60 dilutions of serum from hAd5 S-Fusion + N-ETSD vaccinated mice. (b) The results in the Vero E6 cell SARS-CoV-2 viral infection for mice that showed S-specific antibodies by ELISA also showed high neutralization for mice and very high neutralization for pooled sera (G4 pool, blue line) even compared to COVID-19 convalescent serum. G4 pool – mice with S-specific antibodies; M1, M2, M3, M4 – mouse ID; +C – convalescent serum; and media – media only negative control.

    Techniques Used: Neutralization, Inhibition, Mouse Assay, Infection, Enzyme-linked Immunosorbent Assay, Negative Control

    16) Product Images from "Heterogeneous antibodies against SARS-CoV-2 spike receptor binding domain and nucleocapsid with implications for COVID-19 immunity"

    Article Title: Heterogeneous antibodies against SARS-CoV-2 spike receptor binding domain and nucleocapsid with implications for COVID-19 immunity

    Journal: JCI Insight

    doi: 10.1172/jci.insight.142386

    Comparison of seroconversion in patients with COVID-19 and healthy individuals. ( A ) ELISA with S-RBD protein coating and 1:100 dilution of repeated serum samples of patients with SARS-CoV-2 and healthy individuals. Absorbance normalized to the respective no antigen control for each sample at 450 nm reported. SARS-CoV-2 (blue), n = 88 (from 21 patients); HS 2017–2019 (white), n = 104; HS 2020 (white), n = 308. Arrows list consecutive serum samples evaluated for each case. Inset graphs depict the data separated based on healthy serum collected from 2017 to 2019 (left inset) and 2020 (right inset). ( B ) ELISA with N-protein coating and 1:100 dilution of the first and last serum samples of patients with SARS-CoV-2 and healthy individuals. Absorbance normalized to the respective no antigen control for each sample at 450 nm reported. SARS-CoV-2 (blue), n = 37 (from 21 patients); HS 2017–2019 (white), n = 103; HS 2020 (white), n = 308. Arrows list consecutive serum samples evaluated for each case. Inset graphs depict the data separated based on healthy serum collected from 2017 to 2019 (top inset) and 2020 (bottom inset). ( C ) Pie charts depicting percentage of samples positive for indicated antigens. SARS-CoV-2, n = 21; HS 2017–2019, n = 103; HS 2020, n = 308; non–COVID-19 samples (NCSs), n = 45; HIV, n = 7; all, n = 484.
    Figure Legend Snippet: Comparison of seroconversion in patients with COVID-19 and healthy individuals. ( A ) ELISA with S-RBD protein coating and 1:100 dilution of repeated serum samples of patients with SARS-CoV-2 and healthy individuals. Absorbance normalized to the respective no antigen control for each sample at 450 nm reported. SARS-CoV-2 (blue), n = 88 (from 21 patients); HS 2017–2019 (white), n = 104; HS 2020 (white), n = 308. Arrows list consecutive serum samples evaluated for each case. Inset graphs depict the data separated based on healthy serum collected from 2017 to 2019 (left inset) and 2020 (right inset). ( B ) ELISA with N-protein coating and 1:100 dilution of the first and last serum samples of patients with SARS-CoV-2 and healthy individuals. Absorbance normalized to the respective no antigen control for each sample at 450 nm reported. SARS-CoV-2 (blue), n = 37 (from 21 patients); HS 2017–2019 (white), n = 103; HS 2020 (white), n = 308. Arrows list consecutive serum samples evaluated for each case. Inset graphs depict the data separated based on healthy serum collected from 2017 to 2019 (top inset) and 2020 (bottom inset). ( C ) Pie charts depicting percentage of samples positive for indicated antigens. SARS-CoV-2, n = 21; HS 2017–2019, n = 103; HS 2020, n = 308; non–COVID-19 samples (NCSs), n = 45; HIV, n = 7; all, n = 484.

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Detection of serum binding antibodies against SARS-CoV-2 proteins in patients with PCR-confirmed COVID-19 and healthy samples. ( A ) Timeline of COVID-19 diagnosis/ICU admittance, serum sample collection, and convalescent plasma (CP) administration. Time 0 is defined as day of COVID-19 diagnosis (PCR positive for SARS-CoV-2) and ICU admittance. Blood collections are denoted in gray and CP administration is denoted in pink. Patients were stratified based on current status (recovered, hospitalized, or deceased). Patient 29 from our cohort had symptoms but was PCR negative for SARS-CoV-2; this sample was not included in figures since there was no proof of disease. ( B ) Schematic of SARS-CoV-2 viral structure (top panel) and antigens assayed (bottom panel). S-protein, light orange; envelope protein, yellow; membrane glycoprotein, dark orange; RNA, blue; N-protein, green. Absorbance normalized to the respective no antigen control for each sample at 450 nm plotted for S-RBD (left panel), and N-protein (right panel), antigen coating with the most recent (or only) SARS-CoV-2 samples not treated with CP ( n = 21) and healthy samples collected in 2017–2019 (HS 2017–2019, n = 104 for S-RBD, n = 103 for N-protein) and 2020 (HS 2020, n = 308). Data are presented with each dot representing the mean normalized absorbance for a given serum sample; the red bar depicts the median ± interquartile range of all samples. HS, healthy sample; NC (line), negative control cutoff (see Methods). Kruskal-Wallis with Dunn’s multiple-comparisons test performed. **** P
    Figure Legend Snippet: Detection of serum binding antibodies against SARS-CoV-2 proteins in patients with PCR-confirmed COVID-19 and healthy samples. ( A ) Timeline of COVID-19 diagnosis/ICU admittance, serum sample collection, and convalescent plasma (CP) administration. Time 0 is defined as day of COVID-19 diagnosis (PCR positive for SARS-CoV-2) and ICU admittance. Blood collections are denoted in gray and CP administration is denoted in pink. Patients were stratified based on current status (recovered, hospitalized, or deceased). Patient 29 from our cohort had symptoms but was PCR negative for SARS-CoV-2; this sample was not included in figures since there was no proof of disease. ( B ) Schematic of SARS-CoV-2 viral structure (top panel) and antigens assayed (bottom panel). S-protein, light orange; envelope protein, yellow; membrane glycoprotein, dark orange; RNA, blue; N-protein, green. Absorbance normalized to the respective no antigen control for each sample at 450 nm plotted for S-RBD (left panel), and N-protein (right panel), antigen coating with the most recent (or only) SARS-CoV-2 samples not treated with CP ( n = 21) and healthy samples collected in 2017–2019 (HS 2017–2019, n = 104 for S-RBD, n = 103 for N-protein) and 2020 (HS 2020, n = 308). Data are presented with each dot representing the mean normalized absorbance for a given serum sample; the red bar depicts the median ± interquartile range of all samples. HS, healthy sample; NC (line), negative control cutoff (see Methods). Kruskal-Wallis with Dunn’s multiple-comparisons test performed. **** P

    Techniques Used: Binding Assay, Polymerase Chain Reaction, Negative Control

    17) Product Images from "Evolution of antibody immunity to SARS-CoV-2"

    Article Title: Evolution of antibody immunity to SARS-CoV-2

    Journal: Nature

    doi: 10.1038/s41586-021-03207-w

    SARS-CoV-2 antigen and RNA is detectable in different intestinal segments in several individuals convalescent from COVID-19. a , Immunofluorescence images of biopsy samples in the gastrointestinal tract in different individuals are shown. Staining is for EPCAM (red), DAPI (blue) and SARS-CoV-2 nucleocapsid (green). Samples are derived from intestinal biopsies from four participants (CGI-089, CGI-092, CGI-100 and CGI-106) taken at least three months after COVID-19 infection. Arrows indicate enterocytes with detectable SARS-CoV-2 antigen. Scale bars, 100 μm. The experiments were repeated independently at least twice with similar results. b , Quantification of SARS-CoV-2-positive cells by immunofluorescence. The number of cells staining positive for the N protein of SARS-CoV-2 per mm 2 of intestinal epithelium is shown. The graphs show biopsy samples from the indicated individuals of the duodenum (left) and terminal ileum (right). Black dots represent the number of available biopsy specimen for each individual from the respective intestinal segment (CGI-088, n = 4 duodenal and n = 2 ileal; CGI-089, n = 2 duodenal and n = 2 ileal; CGI-092, n = 6 duodenal and n = 3 ileal; CGI-106, n = 4; CGI-100, n = 4). Boxes represent median values and whiskers the 95% confidence interval. c , d , SARS-CoV-2 viral RNA was visualized in intestinal biopsies of participant CGI-089 ( c ) and CGI-088 ( d ) using in situ hybridization. SARS-CoV-2 genomic RNA (black) and haematoxylin and eosin staining by smFISH–immunohistochemistry technique in the duodenum (left) or terminal ileum (right). Arrows indicate enterocytes with detectable SARS-CoV-2 RNA. e , Pre-COVID-19 control individuals show no detectable SARS-CoV-2 viral RNA in duodenal (left) or ileal (right) biopsies. Scale bars, 100 μm. The experiments were repeated independently three times with similar results.
    Figure Legend Snippet: SARS-CoV-2 antigen and RNA is detectable in different intestinal segments in several individuals convalescent from COVID-19. a , Immunofluorescence images of biopsy samples in the gastrointestinal tract in different individuals are shown. Staining is for EPCAM (red), DAPI (blue) and SARS-CoV-2 nucleocapsid (green). Samples are derived from intestinal biopsies from four participants (CGI-089, CGI-092, CGI-100 and CGI-106) taken at least three months after COVID-19 infection. Arrows indicate enterocytes with detectable SARS-CoV-2 antigen. Scale bars, 100 μm. The experiments were repeated independently at least twice with similar results. b , Quantification of SARS-CoV-2-positive cells by immunofluorescence. The number of cells staining positive for the N protein of SARS-CoV-2 per mm 2 of intestinal epithelium is shown. The graphs show biopsy samples from the indicated individuals of the duodenum (left) and terminal ileum (right). Black dots represent the number of available biopsy specimen for each individual from the respective intestinal segment (CGI-088, n = 4 duodenal and n = 2 ileal; CGI-089, n = 2 duodenal and n = 2 ileal; CGI-092, n = 6 duodenal and n = 3 ileal; CGI-106, n = 4; CGI-100, n = 4). Boxes represent median values and whiskers the 95% confidence interval. c , d , SARS-CoV-2 viral RNA was visualized in intestinal biopsies of participant CGI-089 ( c ) and CGI-088 ( d ) using in situ hybridization. SARS-CoV-2 genomic RNA (black) and haematoxylin and eosin staining by smFISH–immunohistochemistry technique in the duodenum (left) or terminal ileum (right). Arrows indicate enterocytes with detectable SARS-CoV-2 RNA. e , Pre-COVID-19 control individuals show no detectable SARS-CoV-2 viral RNA in duodenal (left) or ileal (right) biopsies. Scale bars, 100 μm. The experiments were repeated independently three times with similar results.

    Techniques Used: Immunofluorescence, Staining, Derivative Assay, Infection, In Situ Hybridization, Immunohistochemistry

    Reactivity of anti-SARS-CoV-2 RBD monoclonal antibodies. a , Graph shows anti-SARS-CoV-2 RBD antibody reactivity. ELISA EC 50 , and 122 selected monoclonal antibodies measured at 6.2 months. Horizontal bars indicate geometric mean. Statistical significance was determined using two-tailed Mann–Whitney U -test. b , EC 50 values for all 52 antibodies that appear at 1.3 and 6.2 months. Average of two or more experiments. Horizontal bars indicate geometric mean. Statistical significance was determined using two-tailed Wilcoxon matched-pairs signed-rank test. c , Surface representation of the RBD with the ACE2-binding footprint indicated as a dotted line and selected residues found in circulating strains (grey) and residues that mediate resistance to class-2 (red) (C144) and −3 (green) (C135) antibodies highlighted as sticks. d , Graphs show ELISA binding curves for C144 (black dashed line) and its clonal relatives obtained after 6.2 months (C050, C051, C052, C053 and C054) (solid lines) binding to wild-type (WT), Q493R, R346S and E484K RBDs. e . Heat map shows log 2 -transformed relative fold change in EC 50 against indicated RBD mutants for 26 antibody clonal pairs obtained at 1.3 and 6.2 months with the most pronounced changes in reactivity. The participant of origin for each antibody pair is indicated above. All experiments were performed at least twice.
    Figure Legend Snippet: Reactivity of anti-SARS-CoV-2 RBD monoclonal antibodies. a , Graph shows anti-SARS-CoV-2 RBD antibody reactivity. ELISA EC 50 , and 122 selected monoclonal antibodies measured at 6.2 months. Horizontal bars indicate geometric mean. Statistical significance was determined using two-tailed Mann–Whitney U -test. b , EC 50 values for all 52 antibodies that appear at 1.3 and 6.2 months. Average of two or more experiments. Horizontal bars indicate geometric mean. Statistical significance was determined using two-tailed Wilcoxon matched-pairs signed-rank test. c , Surface representation of the RBD with the ACE2-binding footprint indicated as a dotted line and selected residues found in circulating strains (grey) and residues that mediate resistance to class-2 (red) (C144) and −3 (green) (C135) antibodies highlighted as sticks. d , Graphs show ELISA binding curves for C144 (black dashed line) and its clonal relatives obtained after 6.2 months (C050, C051, C052, C053 and C054) (solid lines) binding to wild-type (WT), Q493R, R346S and E484K RBDs. e . Heat map shows log 2 -transformed relative fold change in EC 50 against indicated RBD mutants for 26 antibody clonal pairs obtained at 1.3 and 6.2 months with the most pronounced changes in reactivity. The participant of origin for each antibody pair is indicated above. All experiments were performed at least twice.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Two Tailed Test, MANN-WHITNEY, Binding Assay, Transformation Assay

    Neutralization of wild-type and mutant RBDs, C51 alignment and binding projection. a , IC 50 values of shared singlets and shared clones of monoclonal antibodies obtained at the initial 1.3- and 6.2-months follow-up visit, divided by participant ( n = 6 (COV21), n = 13 (COV47), n = 3 (COV57), n = 6 (COV72), n = 15 (COV96), n = 9 (COV107)). Lines connect shared singlets or clones. Monoclonal antibodies with undetectable IC 50 at 1.3 months are plotted at 10 μg ml −1 and are highlighted in red; monoclobal antibodies with improved IC 50 at the 6.2-month follow-up visit are highlighted in green; remaining monoclonal antibodies are shown in black. Statistical significance was determined using two-tailed Wilcoxon matched-pairs signed-rank test. b–f , The normalized relative luminescence values for cell lysates of 293T ACE2 cells 48 h after infection with SARS-CoV-2 pseudovirus containing wild-type RBD or RBD mutants (wild-type, Q493R, E484G and R346S RBD shown in black, red, green and blue, respectively) in the presence of increasing concentrations of monoclonal antibodies obtained at the 1.3-month initial visit (1.3m, dashed lines) and their shared clones or singlets at the 6.2-month follow-up visit (6.2m, continuous lines). Antibody identifiers are as indicated. g , VH and VL amino acid sequence alignment of C144 and derivative antibodies C051, C052, C053 and C054. Germline-encoded residues are highlighted in green. Residues in the proximity of RBD-binding C144 paratope are highlighted in red. h–j , Surface representation of two adjacent ‘down’ RBDs (RBD A and RBD B ) on a spike trimer with the C144 epitope on the RBDs highlighted in cyan and positions of amino acid mutations that accumulated in C052 ( h ), C053 ( i ) and C054 ( j .
    Figure Legend Snippet: Neutralization of wild-type and mutant RBDs, C51 alignment and binding projection. a , IC 50 values of shared singlets and shared clones of monoclonal antibodies obtained at the initial 1.3- and 6.2-months follow-up visit, divided by participant ( n = 6 (COV21), n = 13 (COV47), n = 3 (COV57), n = 6 (COV72), n = 15 (COV96), n = 9 (COV107)). Lines connect shared singlets or clones. Monoclonal antibodies with undetectable IC 50 at 1.3 months are plotted at 10 μg ml −1 and are highlighted in red; monoclobal antibodies with improved IC 50 at the 6.2-month follow-up visit are highlighted in green; remaining monoclonal antibodies are shown in black. Statistical significance was determined using two-tailed Wilcoxon matched-pairs signed-rank test. b–f , The normalized relative luminescence values for cell lysates of 293T ACE2 cells 48 h after infection with SARS-CoV-2 pseudovirus containing wild-type RBD or RBD mutants (wild-type, Q493R, E484G and R346S RBD shown in black, red, green and blue, respectively) in the presence of increasing concentrations of monoclonal antibodies obtained at the 1.3-month initial visit (1.3m, dashed lines) and their shared clones or singlets at the 6.2-month follow-up visit (6.2m, continuous lines). Antibody identifiers are as indicated. g , VH and VL amino acid sequence alignment of C144 and derivative antibodies C051, C052, C053 and C054. Germline-encoded residues are highlighted in green. Residues in the proximity of RBD-binding C144 paratope are highlighted in red. h–j , Surface representation of two adjacent ‘down’ RBDs (RBD A and RBD B ) on a spike trimer with the C144 epitope on the RBDs highlighted in cyan and positions of amino acid mutations that accumulated in C052 ( h ), C053 ( i ) and C054 ( j .

    Techniques Used: Neutralization, Mutagenesis, Binding Assay, Clone Assay, Two Tailed Test, Infection, Sequencing

    Immunofluorescence imaging of intestinal biopsies. a , Immunofluorescence images of human enterocytes stained for EPCAM (red), DAPI (blue) and either ACE2 (green in a , c ) or SARS-CoV-2 N (green in b , d ) in intestinal biopsies taken 92 d after onset of COVID-19 symptoms in participant CGI-088, in the terminal ileum ( a , b ) or duodenum ( c , d ). Regions in white boxes in the right panels of a , c are shown expanded in b , d , respectively. Arrows indicate enterocytes with detectable SARS-CoV-2 antigen. Scale bars, 100 μm. The experiments were repeated independently at least twice with similar results.
    Figure Legend Snippet: Immunofluorescence imaging of intestinal biopsies. a , Immunofluorescence images of human enterocytes stained for EPCAM (red), DAPI (blue) and either ACE2 (green in a , c ) or SARS-CoV-2 N (green in b , d ) in intestinal biopsies taken 92 d after onset of COVID-19 symptoms in participant CGI-088, in the terminal ileum ( a , b ) or duodenum ( c , d ). Regions in white boxes in the right panels of a , c are shown expanded in b , d , respectively. Arrows indicate enterocytes with detectable SARS-CoV-2 antigen. Scale bars, 100 μm. The experiments were repeated independently at least twice with similar results.

    Techniques Used: Immunofluorescence, Imaging, Staining

    Neutralizing activity of anti-SARS-CoV-2 RBD monoclonal antibodies. a , SARS-CoV-2 pseudovirus neutralization assay. IC 50 , and 122 selected antibodies measured at 6.2 months. Antibodies with IC 50 values above 1 μg ml −1 were plotted at 1 μg ml −1 . Mean of two independent experiments. Red bar indicates geometric mean. Statistical significance was determined using two-tailed Mann–Whitney U -test. b , IC 50 values for 52 antibodies appearing at 1.3 and 6.2 months. Red bar indicates geometric mean. Statistical significance was determined using two-tailed Wilcoxon matched-pairs signed-rank test. c , IC 50 values for 5 pairs of monoclonal antibody clonal relatives obtained after 1.3 or 6.2 months for neutralization of wild-type and mutant SARS-CoV-2 pseudovirus. Antibody identifiers of the 1.3-month–6.2-month monoclonal antibody pairs as indicated. Bold styling denotes antibody pairs with substantial increase in neutralizing activity after 6.2 months. d , Graph shows the normalized relative luminescence units (RLU) for cell lysates of 293T cells expressing ACE2, 48 h after infection with SARS-CoV-2 pseudovirus containing wild-type RBD or one of three mutant RBDs (Q493R, E484G and R346S) in the presence of increasing concentrations of one of two monoclonal antibodies C144 (1.3 months) (dashed lines) or C051 (6.2 months) (solid lines). Experiments were performed at least twice. e , C051 binding model. Surface representation of two adjacent ‘down’ RBDs (RBD A and RBD B ) on a spike trimer, with the C144 epitope on the RBDs highlighted in cyan and positions of amino acid mutations that accumulated in C051 compared to the parent antibody C144 highlighted as stick side chains on a Cα atom representation of C051 V H V L . HC, heavy chain; LC, light chain.
    Figure Legend Snippet: Neutralizing activity of anti-SARS-CoV-2 RBD monoclonal antibodies. a , SARS-CoV-2 pseudovirus neutralization assay. IC 50 , and 122 selected antibodies measured at 6.2 months. Antibodies with IC 50 values above 1 μg ml −1 were plotted at 1 μg ml −1 . Mean of two independent experiments. Red bar indicates geometric mean. Statistical significance was determined using two-tailed Mann–Whitney U -test. b , IC 50 values for 52 antibodies appearing at 1.3 and 6.2 months. Red bar indicates geometric mean. Statistical significance was determined using two-tailed Wilcoxon matched-pairs signed-rank test. c , IC 50 values for 5 pairs of monoclonal antibody clonal relatives obtained after 1.3 or 6.2 months for neutralization of wild-type and mutant SARS-CoV-2 pseudovirus. Antibody identifiers of the 1.3-month–6.2-month monoclonal antibody pairs as indicated. Bold styling denotes antibody pairs with substantial increase in neutralizing activity after 6.2 months. d , Graph shows the normalized relative luminescence units (RLU) for cell lysates of 293T cells expressing ACE2, 48 h after infection with SARS-CoV-2 pseudovirus containing wild-type RBD or one of three mutant RBDs (Q493R, E484G and R346S) in the presence of increasing concentrations of one of two monoclonal antibodies C144 (1.3 months) (dashed lines) or C051 (6.2 months) (solid lines). Experiments were performed at least twice. e , C051 binding model. Surface representation of two adjacent ‘down’ RBDs (RBD A and RBD B ) on a spike trimer, with the C144 epitope on the RBDs highlighted in cyan and positions of amino acid mutations that accumulated in C051 compared to the parent antibody C144 highlighted as stick side chains on a Cα atom representation of C051 V H V L . HC, heavy chain; LC, light chain.

    Techniques Used: Activity Assay, Neutralization, Two Tailed Test, MANN-WHITNEY, Mutagenesis, Expressing, Infection, Binding Assay

    Persistent longitudinal changes in the phenotypic landscape of B cells in individuals recovered from COVID-19. a , Global viSNE projection of pooled B cells for all participants pooled shown in background contour plots, with overlaid projections of concatenated controls, and convalescent participants at 1.3 and 6.2 months. b , viSNE projection of pooled B cells for all participants of B cell clusters identified by FlowSOM clustering. Column-scaled z -scores of median fluorescence intensity (MFI) as indicated by cluster and marker. c , Frequency of B cells from each group in FlowSOM clusters indicated. Each circle represents an individual control individual ( n = 20) (grey), convalescent participant at 1.3-month post-infection ( n = 41) (red) or convalescent participant at 6.2 months post-infection ( n = 41) (green). Horizontal bars indicate mean values. Significance determined by two-tailed paired t -test for comparisons between time points within individuals and two-tailed unpaired t -test for comparison between controls and convalescent individuals. d , Individual viSNE projections of indicated protein expression.
    Figure Legend Snippet: Persistent longitudinal changes in the phenotypic landscape of B cells in individuals recovered from COVID-19. a , Global viSNE projection of pooled B cells for all participants pooled shown in background contour plots, with overlaid projections of concatenated controls, and convalescent participants at 1.3 and 6.2 months. b , viSNE projection of pooled B cells for all participants of B cell clusters identified by FlowSOM clustering. Column-scaled z -scores of median fluorescence intensity (MFI) as indicated by cluster and marker. c , Frequency of B cells from each group in FlowSOM clusters indicated. Each circle represents an individual control individual ( n = 20) (grey), convalescent participant at 1.3-month post-infection ( n = 41) (red) or convalescent participant at 6.2 months post-infection ( n = 41) (green). Horizontal bars indicate mean values. Significance determined by two-tailed paired t -test for comparisons between time points within individuals and two-tailed unpaired t -test for comparison between controls and convalescent individuals. d , Individual viSNE projections of indicated protein expression.

    Techniques Used: Fluorescence, Marker, Infection, Two Tailed Test, Expressing

    SARS-CoV-2 antigen in human enterocytes in the gastrointestinal tract at three months after COVID-19 diagnosis, and pre-COVID-19 control individuals without detectable SARS-CoV-2 antigen. a–j , Immunofluorescence images of human gastrointestinal tissue are shown. Staining is for EPCAM (red), DAPI (blue) and SARS-CoV-2 nucleocapsid (green) Samples are derived from intestinal biopsies in the gastrointestinal tract as indicated. a–h ) taken 92 d after the onset of COVID-19 symptoms. i , j , Biopsy taken 27 months before the onset of COVID-19 symptoms from participant CGI-088. Arrows indicate enterocytes with detectable SARS-CoV-2 antigen. Isotype and no-primary controls for each tissue are shown in the right two columns. Scale bars, 100 μm. k , l , Immunofluorescence images of biopsy samples in the gastrointestinal tract (ileal ( k ) and duodenal ( l )) obtained from ten preCOVID-19 control individuals between January 2018 and October 2019 are shown. Staining is for EPCAM (red), DAPI (blue) and SARS-CoV-2 nucleocapsid (green). Scale bars, 100 μm. All experiments were repeated independently at least twice with similar results.
    Figure Legend Snippet: SARS-CoV-2 antigen in human enterocytes in the gastrointestinal tract at three months after COVID-19 diagnosis, and pre-COVID-19 control individuals without detectable SARS-CoV-2 antigen. a–j , Immunofluorescence images of human gastrointestinal tissue are shown. Staining is for EPCAM (red), DAPI (blue) and SARS-CoV-2 nucleocapsid (green) Samples are derived from intestinal biopsies in the gastrointestinal tract as indicated. a–h ) taken 92 d after the onset of COVID-19 symptoms. i , j , Biopsy taken 27 months before the onset of COVID-19 symptoms from participant CGI-088. Arrows indicate enterocytes with detectable SARS-CoV-2 antigen. Isotype and no-primary controls for each tissue are shown in the right two columns. Scale bars, 100 μm. k , l , Immunofluorescence images of biopsy samples in the gastrointestinal tract (ileal ( k ) and duodenal ( l )) obtained from ten preCOVID-19 control individuals between January 2018 and October 2019 are shown. Staining is for EPCAM (red), DAPI (blue) and SARS-CoV-2 nucleocapsid (green). Scale bars, 100 μm. All experiments were repeated independently at least twice with similar results.

    Techniques Used: Immunofluorescence, Staining, Derivative Assay

    Plasma antibody dynamics against SARS-CoV-2. a–d , Results of ELISAs measuring plasma reactivity to RBD ( a , b , c ) and N protein ( d ) at the initial 1.3- and 6.2-month follow-up visit, respectively. a , Anti-RBD IgM. b , Anti-RBD IgG. c , Anti-RBD IgA d , Anti-N IgG. The normalized area under the curve (AUC) values for 87 individuals are shown in a – d . e , Relative change in plasma antibody levels between 1.3 and 6.2 months for anti-RBD IgM, IgG, IgA and anti-N IgG in all 87 individuals. f – i , Relative change in antibody levels between 1.3 and 6.2 months plotted against the corresponding antibody levels at 1.3 months. f , Anti-RBD IgM. r = −0.83, P
    Figure Legend Snippet: Plasma antibody dynamics against SARS-CoV-2. a–d , Results of ELISAs measuring plasma reactivity to RBD ( a , b , c ) and N protein ( d ) at the initial 1.3- and 6.2-month follow-up visit, respectively. a , Anti-RBD IgM. b , Anti-RBD IgG. c , Anti-RBD IgA d , Anti-N IgG. The normalized area under the curve (AUC) values for 87 individuals are shown in a – d . e , Relative change in plasma antibody levels between 1.3 and 6.2 months for anti-RBD IgM, IgG, IgA and anti-N IgG in all 87 individuals. f – i , Relative change in antibody levels between 1.3 and 6.2 months plotted against the corresponding antibody levels at 1.3 months. f , Anti-RBD IgM. r = −0.83, P

    Techniques Used:

    Related Articles

    Mouse Assay:

    Article Title: Original antigenic sin responses to heterologous Betacoronavirus spike proteins are observed in mice following intramuscular administration, but are not apparent in children following SARS-CoV-2 infection
    Article Snippet: .. One group of five Balb/c mice was primed and boosted at 21 days IM with 10 µg SARS-CoV-2 nucleocapsid protein (SinoBiological, 40588-V08B) in 50 µl with alum (Alhydrogel adjuvant 2%, Invivogen). .. The purpose of this was to generate polyclonal antiserum to the nucleocapsid protein early during the COVID-19 pandemic, which was not a part of the present study.

    Recombinant:

    Article Title: A SARS-CoV-2 antibody curbs viral nucleocapsid protein-induced complement hyperactivation
    Article Snippet: Recombinant SARS-CoV-2 S-ECD and N proteins Recombinant SARS-CoV-2 S protein (extracellular domain of the S protein (ECD) with His and FLAG Tags, Z03481) was purchased from GenScript. .. Recombinant SARS-CoV-2 full-length N protein with a CT 6x His tag (His tag, 40588-V08B) was purchased from Sino Biological. .. The SARS-CoV-2 N protein expression plasmid pET-28a (SARS-CoV-2 N-FL) was a gift from the Guangdong Medical Laboratory Animal Center.

    Article Title: COVID-19 Patients Upregulate Toll-like Receptor 4-mediated Inflammatory Signaling That Mimics Bacterial Sepsis
    Article Snippet: Human MDMs were prepared by culturing peripheral blood monocytes for 4 days in the presence of 4 ng/mL human macrophage colony-stimulating factor (R & D Systems, Minneapolis, MN, USA). .. SARS-CoV-2 (2019-nCoV) NC-His recombinant protein (cat. No. 40588-V08B), Spike S1-His recombinant protein (cat. No. 40591-V08H), Spike S2 extracellular domain (ECD)-His recombinant protein (cat. No. 40590-V08B), and Spike RBD-His recombinant protein (cat. No. 40592-V08H) were purchased from Sino Biological, Beijing, China. .. RNA extraction and RT-qPCR Total RNA from PBMCs or MDMs was extracted using QIAzol lysis reagent (Qiagen, Hilden, Germany) and miRNeasy Mini Kits (Qiagen) according to the manufacturer's instructions, followed by RNA quantitation. cDNA from total RNA was synthesized using the reverse transcription master premix (ELPIS Biotech, Daejeon, Korea).

    Article Title: GCG inhibits SARS-CoV-2 replication by disrupting the liquid phase condensation of its nucleocapsid protein
    Article Snippet: Briefly, GCG was conjugated with cyanogen bromide (CNBr)-activated agarose beads (C500099, Sangon Biotech). .. The recombinant N protein (40588-V08B) was from Sino-Biological. .. A549-hACE2-Flag cells were transfected with pcDNA3.0-Flag-N for 24 h and then lysed with lysis buffer (20 mM Tris-HCl, pH 7.5; 0.5% Nonidet P-40; 250 mM NaCl; 3 mM EDTA and 3 mM EGTA) containing complete protease inhibitor cocktail (04693132001, Roche), followed by centrifugation at 20,000 × g for 20 min at 4 °C.

    Article Title: Development of a Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of SARS-CoV-2 Spike Glycoprotein and Nucleoprotein
    Article Snippet: Purified proteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis to ensure purity and appropriate molecular weights. .. SARS-CoV-2 nucleocapsid-His recombinant protein was purchased from Sino Biological (100 μg, 40588-V08B) The protein was difficult to dissolve and required treatment before S-trap preparation below with 40 μL dimethyl sulfoxide (276855—100 mL, Sigma), 126 μL 1% TFA, and 100 μL 1× S-Trap lysis buffer (5% SDS, 50 mM TEAB, pH adjusted to 7.55 using 12% phosphoric acid). ..

    Article Title: SARS-CoV-2 RapidPlex: A Graphene-Based Multiplexed Telemedicine Platform for Rapid and Low-Cost COVID-19 Diagnosis and Monitoring
    Article Snippet: CRP polyclonal antibody labeled with horseradish peroxidase (HRP) (PA1-28329) and 3,3′,5,5′-tetramethylbenzidine (TMB) colorimetric substrate was purchased from Invitrogen. .. Mouse NP monoclonal antibody (mAb) (40143-MM05), SARS-CoV-2 NP antigen (40588-V08B), SARS-CoV/SARS-CoV-2 nucleocapsid antibody, rabbit mAb (40143-R001), SARS-CoV NP antigen (HCoV-OC43; 40643-V07E), SARS-CoV-2 Spike S1-His recombinant protein (HPLC-verified) (40591-V08H), and SARS-CoV Spike S1 protein (S1 subunit, His tag) (40150-V08B1) were purchased from Sino Biological. ..

    Multiplex Assay:

    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). ..

    other:

    Article Title: The Characterization of Disease Severity Associated IgG Subclasses Response in COVID-19 Patients
    Article Snippet: ProteinsSARS-CoV-2 Spike Protein (S1+S2) (cat# 40589-VO8B1), SARS-CoV-2 Spike RBD Protein (cat# 40592-V08B), SARS-CoV-2 Nucleocapsid Protein (cat# 40588-V08B) were purchased from Sino Biological (China).

    Lysis:

    Article Title: Development of a Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of SARS-CoV-2 Spike Glycoprotein and Nucleoprotein
    Article Snippet: Purified proteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis to ensure purity and appropriate molecular weights. .. SARS-CoV-2 nucleocapsid-His recombinant protein was purchased from Sino Biological (100 μg, 40588-V08B) The protein was difficult to dissolve and required treatment before S-trap preparation below with 40 μL dimethyl sulfoxide (276855—100 mL, Sigma), 126 μL 1% TFA, and 100 μL 1× S-Trap lysis buffer (5% SDS, 50 mM TEAB, pH adjusted to 7.55 using 12% phosphoric acid). ..

    High Performance Liquid Chromatography:

    Article Title: SARS-CoV-2 RapidPlex: A Graphene-Based Multiplexed Telemedicine Platform for Rapid and Low-Cost COVID-19 Diagnosis and Monitoring
    Article Snippet: CRP polyclonal antibody labeled with horseradish peroxidase (HRP) (PA1-28329) and 3,3′,5,5′-tetramethylbenzidine (TMB) colorimetric substrate was purchased from Invitrogen. .. Mouse NP monoclonal antibody (mAb) (40143-MM05), SARS-CoV-2 NP antigen (40588-V08B), SARS-CoV/SARS-CoV-2 nucleocapsid antibody, rabbit mAb (40143-R001), SARS-CoV NP antigen (HCoV-OC43; 40643-V07E), SARS-CoV-2 Spike S1-His recombinant protein (HPLC-verified) (40591-V08H), and SARS-CoV Spike S1 protein (S1 subunit, His tag) (40150-V08B1) were purchased from Sino Biological. ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96
    Sino Biological sars cov 2 2019 ncov nucleocapsid his recombinant protein covid 19 nucleocapsid research
    GCG suppresses <t>SARS-CoV-2</t> replication. a , b Immunofluorescence analysis of N protein in A549-hACE2-Flag cells infected with SARS-CoV-2 for 24 h ( a ). The percentage of cells with N protein foci was quantified, n = 8 biologically independent samples, 20 randomly selected views were analyzed in each sample ( b ). Scale bar, 10 μm. c 3D images were obtained by Zeiss LSM 880 confocal microscope and reconstituted by Volocity 6.1.1 . d , e The inhibitory effect of GCG on the replication of SARS-CoV-2, n = 6 biologically independent samples ( d ). IC 50 was calculated, n = 5 biologically independent samples ( e ). The infection was performed after 1-h pretreatment of GCG. f , g Representative immunofluorescent images showed the inhibitory effect of GCG on SARS-CoV-2 N protein. 3D images were obtained by Zeiss LSM 880 confocal microscope and reconstituted by Volocity 6.1.1 ( f ). Violin plots showing foci of cells ( n = 50 biologically independent cells) from each group, lines within the plots, with 25th, 50th, and 75th percentiles marked ( g ). h Cells were infected with SARS-CoV-2 for 1 h followed by 24-h GCG treatment, n = 3 biologically independent samples. Representative images were shown. SARS-CoV-2 was used at an MOI of 1. Hoechst (blue), nuclear staining ( a , c , f ). Error bars, mean with s.d. ( b , d , e , g , h ). Two-tailed unpaired Student’s t -test, * P
    Sars Cov 2 2019 Ncov Nucleocapsid His Recombinant Protein Covid 19 Nucleocapsid Research, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sars cov 2 2019 ncov nucleocapsid his recombinant protein covid 19 nucleocapsid research/product/Sino Biological
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 2019 ncov nucleocapsid his recombinant protein covid 19 nucleocapsid research - by Bioz Stars, 2021-07
    96/100 stars
      Buy from Supplier

    Image Search Results


    GCG suppresses SARS-CoV-2 replication. a , b Immunofluorescence analysis of N protein in A549-hACE2-Flag cells infected with SARS-CoV-2 for 24 h ( a ). The percentage of cells with N protein foci was quantified, n = 8 biologically independent samples, 20 randomly selected views were analyzed in each sample ( b ). Scale bar, 10 μm. c 3D images were obtained by Zeiss LSM 880 confocal microscope and reconstituted by Volocity 6.1.1 . d , e The inhibitory effect of GCG on the replication of SARS-CoV-2, n = 6 biologically independent samples ( d ). IC 50 was calculated, n = 5 biologically independent samples ( e ). The infection was performed after 1-h pretreatment of GCG. f , g Representative immunofluorescent images showed the inhibitory effect of GCG on SARS-CoV-2 N protein. 3D images were obtained by Zeiss LSM 880 confocal microscope and reconstituted by Volocity 6.1.1 ( f ). Violin plots showing foci of cells ( n = 50 biologically independent cells) from each group, lines within the plots, with 25th, 50th, and 75th percentiles marked ( g ). h Cells were infected with SARS-CoV-2 for 1 h followed by 24-h GCG treatment, n = 3 biologically independent samples. Representative images were shown. SARS-CoV-2 was used at an MOI of 1. Hoechst (blue), nuclear staining ( a , c , f ). Error bars, mean with s.d. ( b , d , e , g , h ). Two-tailed unpaired Student’s t -test, * P

    Journal: Nature Communications

    Article Title: GCG inhibits SARS-CoV-2 replication by disrupting the liquid phase condensation of its nucleocapsid protein

    doi: 10.1038/s41467-021-22297-8

    Figure Lengend Snippet: GCG suppresses SARS-CoV-2 replication. a , b Immunofluorescence analysis of N protein in A549-hACE2-Flag cells infected with SARS-CoV-2 for 24 h ( a ). The percentage of cells with N protein foci was quantified, n = 8 biologically independent samples, 20 randomly selected views were analyzed in each sample ( b ). Scale bar, 10 μm. c 3D images were obtained by Zeiss LSM 880 confocal microscope and reconstituted by Volocity 6.1.1 . d , e The inhibitory effect of GCG on the replication of SARS-CoV-2, n = 6 biologically independent samples ( d ). IC 50 was calculated, n = 5 biologically independent samples ( e ). The infection was performed after 1-h pretreatment of GCG. f , g Representative immunofluorescent images showed the inhibitory effect of GCG on SARS-CoV-2 N protein. 3D images were obtained by Zeiss LSM 880 confocal microscope and reconstituted by Volocity 6.1.1 ( f ). Violin plots showing foci of cells ( n = 50 biologically independent cells) from each group, lines within the plots, with 25th, 50th, and 75th percentiles marked ( g ). h Cells were infected with SARS-CoV-2 for 1 h followed by 24-h GCG treatment, n = 3 biologically independent samples. Representative images were shown. SARS-CoV-2 was used at an MOI of 1. Hoechst (blue), nuclear staining ( a , c , f ). Error bars, mean with s.d. ( b , d , e , g , h ). Two-tailed unpaired Student’s t -test, * P

    Article Snippet: The recombinant N protein (40588-V08B) was from Sino-Biological.

    Techniques: Immunofluorescence, Infection, Microscopy, Staining, Two Tailed Test

    RNA triggers the LLPS of N protein. a Schematic drawing of SARS-CoV-2. b IDR scores of 29 proteins encoded by SARS-CoV-2 genome. FUS and mEGFP are positive and negative controls, respectively. IUPred2 and ANCHOR2 were used as prediction tools. c Time-lapse imaging of N-mEGFP protein (20 μM) in the presence of Cy5-labeled 60-nt vRNA (100 ng/μl), scale bar, 10 μm. d Representative fluorescent images of N-mEGFP-vRNA (60 nt) condensates fusion from a time-lapse movie, scale bar, 3 μm. e – g LLPS of N-mEGFP protein (20 μM) in the presence of indicated concentrations of 60-nt vRNA, scale bar, 10 μm ( e ). The partition coefficient of fluorescence intensity per droplet ( f ) and the partition coefficient of total fluorescence intensity in each view ( g ) were calculated. From left to right, n = 209, 1170, 1026, 1170 droplets ( f ) from 10 randomly selected views ( g ). h , i FRAP analysis of vRNA-induced liquid droplets of N-mEGFP protein, scale bar, 2 μm ( h ), and quantification of fluorescence intensity recovery of a photobleached N-mEGFP protein, n = 3 biologically independent experiments ( i ). The white dotted circle in h indicated the region of photobleaching. 20 μM N-mEGFP protein and 100 ng/μl 60-nt vRNA were used. Error bars, mean with s.d. ( f , g , i ). Two-tailed unpaired Student’s t -test ( f , g ), **** P

    Journal: Nature Communications

    Article Title: GCG inhibits SARS-CoV-2 replication by disrupting the liquid phase condensation of its nucleocapsid protein

    doi: 10.1038/s41467-021-22297-8

    Figure Lengend Snippet: RNA triggers the LLPS of N protein. a Schematic drawing of SARS-CoV-2. b IDR scores of 29 proteins encoded by SARS-CoV-2 genome. FUS and mEGFP are positive and negative controls, respectively. IUPred2 and ANCHOR2 were used as prediction tools. c Time-lapse imaging of N-mEGFP protein (20 μM) in the presence of Cy5-labeled 60-nt vRNA (100 ng/μl), scale bar, 10 μm. d Representative fluorescent images of N-mEGFP-vRNA (60 nt) condensates fusion from a time-lapse movie, scale bar, 3 μm. e – g LLPS of N-mEGFP protein (20 μM) in the presence of indicated concentrations of 60-nt vRNA, scale bar, 10 μm ( e ). The partition coefficient of fluorescence intensity per droplet ( f ) and the partition coefficient of total fluorescence intensity in each view ( g ) were calculated. From left to right, n = 209, 1170, 1026, 1170 droplets ( f ) from 10 randomly selected views ( g ). h , i FRAP analysis of vRNA-induced liquid droplets of N-mEGFP protein, scale bar, 2 μm ( h ), and quantification of fluorescence intensity recovery of a photobleached N-mEGFP protein, n = 3 biologically independent experiments ( i ). The white dotted circle in h indicated the region of photobleaching. 20 μM N-mEGFP protein and 100 ng/μl 60-nt vRNA were used. Error bars, mean with s.d. ( f , g , i ). Two-tailed unpaired Student’s t -test ( f , g ), **** P

    Article Snippet: The recombinant N protein (40588-V08B) was from Sino-Biological.

    Techniques: Imaging, Labeling, Fluorescence, Two Tailed Test

    NR203K/G204R gained greater ability to undergo RNA-induced LLPS. a Distribution of N gene variants among 100,849 SARS-CoV-2 genomes obtained from GISAID database. Colors indicated the nucleotide variability numbers from 100,849 genomes. The high-frequency trio-nucleotide polymorphism variant (GGG-to-AAC) is shown. b Coomassie brilliant blue-stained SDS-PAGE gel of purified variants of N-mEGFP protein. c – e LLPS of different N-mEGFP variants, in the presence of 50 ng/μl 60-nt vRNA ( c ). The partition coefficient of fluorescence intensity per droplet ( d ) and the partition coefficient of total fluorescence intensity in each view ( e ) were calculated. From left to right, n = 1232, 803, 897, 431 droplets ( d ) from 10 randomly selected views ( e ). f , g Time-lapse imaging of N R203/G204 -mEGFP and N R203K/G204R -mEGFP proteins (20 μM) in the presence of Cy5-labeled 60-nt vRNA (40 ng/μl) ( f ), and the partition coefficient ( n = 8 randomly selected views) of total fluorescence intensity in each view ( g ). Scale bars, 10 μm ( c , f ). Error bars, mean with s.d. ( d , e ) and mean with s.e.m. ( g ). Two-tailed unpaired Student’s t -test ( d , e ), **** P

    Journal: Nature Communications

    Article Title: GCG inhibits SARS-CoV-2 replication by disrupting the liquid phase condensation of its nucleocapsid protein

    doi: 10.1038/s41467-021-22297-8

    Figure Lengend Snippet: NR203K/G204R gained greater ability to undergo RNA-induced LLPS. a Distribution of N gene variants among 100,849 SARS-CoV-2 genomes obtained from GISAID database. Colors indicated the nucleotide variability numbers from 100,849 genomes. The high-frequency trio-nucleotide polymorphism variant (GGG-to-AAC) is shown. b Coomassie brilliant blue-stained SDS-PAGE gel of purified variants of N-mEGFP protein. c – e LLPS of different N-mEGFP variants, in the presence of 50 ng/μl 60-nt vRNA ( c ). The partition coefficient of fluorescence intensity per droplet ( d ) and the partition coefficient of total fluorescence intensity in each view ( e ) were calculated. From left to right, n = 1232, 803, 897, 431 droplets ( d ) from 10 randomly selected views ( e ). f , g Time-lapse imaging of N R203/G204 -mEGFP and N R203K/G204R -mEGFP proteins (20 μM) in the presence of Cy5-labeled 60-nt vRNA (40 ng/μl) ( f ), and the partition coefficient ( n = 8 randomly selected views) of total fluorescence intensity in each view ( g ). Scale bars, 10 μm ( c , f ). Error bars, mean with s.d. ( d , e ) and mean with s.e.m. ( g ). Two-tailed unpaired Student’s t -test ( d , e ), **** P

    Article Snippet: The recombinant N protein (40588-V08B) was from Sino-Biological.

    Techniques: Variant Assay, Staining, SDS Page, Purification, Fluorescence, Imaging, Labeling, Two Tailed Test

    Antibody nCoV396 compromises SARS-CoV-2 N protein-induced complement hyperactivation. a Flow scheme of the SARS-CoV-2 N protein and nCoV396 influencing the protease activity of MASP-2 in the serum from donors. b Serum-01 to 06 are used to compare the MASP-2 activity to C2 of serum sample with normal C3 (Health-110,113,117, n = 3) and serum sample with abnormal C3 (Patient-81,123,130, n = 3), and the Michaelis–Menten curves of are presented as mean (three groups of above health serum alone with patient’s serum paired data are shown in Supplementary Fig. 8a ). c The Michaelis–Menten curve of N protein-induced excessive cleavage of C2 in the presence of recombinant MASP-2 in vitro. The reaction system without N protein, the increase of N protein concentration and negative control protein (ENL) expressed in E. coli are presenting. The Michaelis–Menten curve shows the effect of increasing the N protein concentration ( d ) and antibody concentration ( f ) on the substrate C2 cleavage of MAPS-2 in the Serum-07 and Serum-08. e A Hanes plot where C2 concentration/V0 is plotted against C2 concentration with the addition of 5 μM N protein. b – d , f All samples were performed in triplicates and mean were presented. g Five serum sample from biologically independent donors ( n = 5) with abnormal serologic C3 values. (Serum-08 to −12). And we used Michaelis–Menten equation to calculate the V max (with experimental data from Fig. 4f (Serum-08), Supplementary Fig. 5b (Serum-09 to −11), and Supplementary Fig. 7a (Serum-12)). Each sample was performed in triplicates and mean values ± SEM of V max are presented. Two-sided Kruskal–Wallis test with Dunnett’s multiple comparisons test was used for comparing the V max of groups. The significant reference is 0.05.

    Journal: Nature Communications

    Article Title: A SARS-CoV-2 antibody curbs viral nucleocapsid protein-induced complement hyperactivation

    doi: 10.1038/s41467-021-23036-9

    Figure Lengend Snippet: Antibody nCoV396 compromises SARS-CoV-2 N protein-induced complement hyperactivation. a Flow scheme of the SARS-CoV-2 N protein and nCoV396 influencing the protease activity of MASP-2 in the serum from donors. b Serum-01 to 06 are used to compare the MASP-2 activity to C2 of serum sample with normal C3 (Health-110,113,117, n = 3) and serum sample with abnormal C3 (Patient-81,123,130, n = 3), and the Michaelis–Menten curves of are presented as mean (three groups of above health serum alone with patient’s serum paired data are shown in Supplementary Fig. 8a ). c The Michaelis–Menten curve of N protein-induced excessive cleavage of C2 in the presence of recombinant MASP-2 in vitro. The reaction system without N protein, the increase of N protein concentration and negative control protein (ENL) expressed in E. coli are presenting. The Michaelis–Menten curve shows the effect of increasing the N protein concentration ( d ) and antibody concentration ( f ) on the substrate C2 cleavage of MAPS-2 in the Serum-07 and Serum-08. e A Hanes plot where C2 concentration/V0 is plotted against C2 concentration with the addition of 5 μM N protein. b – d , f All samples were performed in triplicates and mean were presented. g Five serum sample from biologically independent donors ( n = 5) with abnormal serologic C3 values. (Serum-08 to −12). And we used Michaelis–Menten equation to calculate the V max (with experimental data from Fig. 4f (Serum-08), Supplementary Fig. 5b (Serum-09 to −11), and Supplementary Fig. 7a (Serum-12)). Each sample was performed in triplicates and mean values ± SEM of V max are presented. Two-sided Kruskal–Wallis test with Dunnett’s multiple comparisons test was used for comparing the V max of groups. The significant reference is 0.05.

    Article Snippet: Recombinant SARS-CoV-2 full-length N protein with a CT 6x His tag (His tag, 40588-V08B) was purchased from Sino Biological.

    Techniques: Activity Assay, Recombinant, In Vitro, Protein Concentration, Negative Control, Concentration Assay

    Complex structure of mAb nCoV396 with SARS-CoV-2 N-NTD. a Overall structure of the mAb nCoV396-SARS-CoV-2 N-NTD complex. The light chain (pink) and heavy chain (blue) of mAb nCoV396 are illustrated with the ribbon representation. SARS-CoV-2 N-NTD is illustrated with electrostatics surface, in which blue denotes a positive charge potential while red indicates a negative charge potential. b The N-NTD epitope recognized by mAb nCoV396. The interacting residues of N-NTD and nCoV396 are highlighted with the stick representation. Recognition of Q163 ( c ), K169 ( d ), and L167 ( e ) in N-NTD by mAb nCoV396. The dashed blue line represents hydrogen bonds. Hydrophobic interactions are illustrated with the dot representation. f Conformational changes of N-NTD upon mAb nCoV396 binding. The apo structure of N-NTD is colored with gray. Antibody-bound N-NTD is colored green. The N-terminus and C-terminus of the N-NTD are labeled with circles. mAb nCoV396 is illustrated with surface representation. All figures were prepared by Pymol.

    Journal: Nature Communications

    Article Title: A SARS-CoV-2 antibody curbs viral nucleocapsid protein-induced complement hyperactivation

    doi: 10.1038/s41467-021-23036-9

    Figure Lengend Snippet: Complex structure of mAb nCoV396 with SARS-CoV-2 N-NTD. a Overall structure of the mAb nCoV396-SARS-CoV-2 N-NTD complex. The light chain (pink) and heavy chain (blue) of mAb nCoV396 are illustrated with the ribbon representation. SARS-CoV-2 N-NTD is illustrated with electrostatics surface, in which blue denotes a positive charge potential while red indicates a negative charge potential. b The N-NTD epitope recognized by mAb nCoV396. The interacting residues of N-NTD and nCoV396 are highlighted with the stick representation. Recognition of Q163 ( c ), K169 ( d ), and L167 ( e ) in N-NTD by mAb nCoV396. The dashed blue line represents hydrogen bonds. Hydrophobic interactions are illustrated with the dot representation. f Conformational changes of N-NTD upon mAb nCoV396 binding. The apo structure of N-NTD is colored with gray. Antibody-bound N-NTD is colored green. The N-terminus and C-terminus of the N-NTD are labeled with circles. mAb nCoV396 is illustrated with surface representation. All figures were prepared by Pymol.

    Article Snippet: Recombinant SARS-CoV-2 full-length N protein with a CT 6x His tag (His tag, 40588-V08B) was purchased from Sino Biological.

    Techniques: Binding Assay, Labeling

    Acquisition and characterization of antibodies. Serum antibody titers of six SARS-CoV-2 convalescent patients and a healthy person (ZH0081, non-COVID-19) to the SARS-CoV-2 S ( a ) and N ( b ) proteins measured by ELISA. All samples were performed in triplicates and mean were presented. Sorting of single plasma cells ( c ) with CD38 and CD27 double-positive B cells. d To minimize false positives, each of the S1 and N proteins labeled with Phycoerythrin-canin7 (PE-Cy7) and Brilliant Violet (BV421) was used to sort antigen-specific memory B cells by FACS. e Percentage of different isotypes, VH and VL gene families of 32 isolated N-reactive antibodies. f Number of mutations in nucleotides and amino acids in VH and VL (Vκ and Vλ) of 32 N-reactive antibodies and eight S-reactive antibodies ( g ). Length of the 32 N-reactive antibodies ( h ) and eight S-reactive antibodies ( i ) in H-CDR3. f – i Data are presented as dot and mean values.

    Journal: Nature Communications

    Article Title: A SARS-CoV-2 antibody curbs viral nucleocapsid protein-induced complement hyperactivation

    doi: 10.1038/s41467-021-23036-9

    Figure Lengend Snippet: Acquisition and characterization of antibodies. Serum antibody titers of six SARS-CoV-2 convalescent patients and a healthy person (ZH0081, non-COVID-19) to the SARS-CoV-2 S ( a ) and N ( b ) proteins measured by ELISA. All samples were performed in triplicates and mean were presented. Sorting of single plasma cells ( c ) with CD38 and CD27 double-positive B cells. d To minimize false positives, each of the S1 and N proteins labeled with Phycoerythrin-canin7 (PE-Cy7) and Brilliant Violet (BV421) was used to sort antigen-specific memory B cells by FACS. e Percentage of different isotypes, VH and VL gene families of 32 isolated N-reactive antibodies. f Number of mutations in nucleotides and amino acids in VH and VL (Vκ and Vλ) of 32 N-reactive antibodies and eight S-reactive antibodies ( g ). Length of the 32 N-reactive antibodies ( h ) and eight S-reactive antibodies ( i ) in H-CDR3. f – i Data are presented as dot and mean values.

    Article Snippet: Recombinant SARS-CoV-2 full-length N protein with a CT 6x His tag (His tag, 40588-V08B) was purchased from Sino Biological.

    Techniques: Enzyme-linked Immunosorbent Assay, Labeling, FACS, Isolation

    Sequence coverage and proteotypic target peptide selection for development of a PRM assay for the SARS CoV-2 Spike protein and NP. (Top panel) Diagram of SARS CoV-2 recombinant spike glycoprotein showing the location of NTD, RBD, fusion peptide and heptad repeats 1 and 2, and the protease cleavage sites, His and Strep tags. The amino acid sequence is given below. Glycosylation sites are indicated in green. (Bottom panel) Diagram of SARS CoV-2 recombinant NP showing intrinsically disordered regions, RNA binding and dimerization regions. Phosphorylation sites (S) are indicated in yellow. Bold italics indicate sites where sequence coverage was not obtained. Peptides monitored in the spectral library are boxed, peptides selected for the final PRM assay are boxed and indicated in red text. Overall 97.1% of the spike protein and 77.2% of the NP sequence was obtained from the DDA analysis.

    Journal: Analytical Chemistry

    Article Title: Development of a Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of SARS-CoV-2 Spike Glycoprotein and Nucleoprotein

    doi: 10.1021/acs.analchem.0c02288

    Figure Lengend Snippet: Sequence coverage and proteotypic target peptide selection for development of a PRM assay for the SARS CoV-2 Spike protein and NP. (Top panel) Diagram of SARS CoV-2 recombinant spike glycoprotein showing the location of NTD, RBD, fusion peptide and heptad repeats 1 and 2, and the protease cleavage sites, His and Strep tags. The amino acid sequence is given below. Glycosylation sites are indicated in green. (Bottom panel) Diagram of SARS CoV-2 recombinant NP showing intrinsically disordered regions, RNA binding and dimerization regions. Phosphorylation sites (S) are indicated in yellow. Bold italics indicate sites where sequence coverage was not obtained. Peptides monitored in the spectral library are boxed, peptides selected for the final PRM assay are boxed and indicated in red text. Overall 97.1% of the spike protein and 77.2% of the NP sequence was obtained from the DDA analysis.

    Article Snippet: SARS-CoV-2 nucleocapsid-His recombinant protein was purchased from Sino Biological (100 μg, 40588-V08B) The protein was difficult to dissolve and required treatment before S-trap preparation below with 40 μL dimethyl sulfoxide (276855—100 mL, Sigma), 126 μL 1% TFA, and 100 μL 1× S-Trap lysis buffer (5% SDS, 50 mM TEAB, pH adjusted to 7.55 using 12% phosphoric acid).

    Techniques: Sequencing, Selection, Recombinant, RNA Binding Assay

    Chromatograms and calibration curves for two best target peptides used in the PRM assay for SARS CoV-2 spike protein and nuceloprotein. The summed area under curve values for the top four transitions of each peptide were taken to generate calibration curves for quantitation. The right panels display chromatograms obtained for each of transitions shown in different colors for (A) DQVILLNK (NP) and (B) FQTLLALHR (S). Three technical replicates were run on two separate days. The chromatograms on the left of each panel show a low and high standard from the SARS CoV-2 S and NP in a mucin background. Calibration curves were constructed from the PRM data (top right) and zoomed in (bottom right) displaying mean values at the low end of the curve to show the LOD (left dotted line) and LOQ (right dotted line).

    Journal: Analytical Chemistry

    Article Title: Development of a Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of SARS-CoV-2 Spike Glycoprotein and Nucleoprotein

    doi: 10.1021/acs.analchem.0c02288

    Figure Lengend Snippet: Chromatograms and calibration curves for two best target peptides used in the PRM assay for SARS CoV-2 spike protein and nuceloprotein. The summed area under curve values for the top four transitions of each peptide were taken to generate calibration curves for quantitation. The right panels display chromatograms obtained for each of transitions shown in different colors for (A) DQVILLNK (NP) and (B) FQTLLALHR (S). Three technical replicates were run on two separate days. The chromatograms on the left of each panel show a low and high standard from the SARS CoV-2 S and NP in a mucin background. Calibration curves were constructed from the PRM data (top right) and zoomed in (bottom right) displaying mean values at the low end of the curve to show the LOD (left dotted line) and LOQ (right dotted line).

    Article Snippet: SARS-CoV-2 nucleocapsid-His recombinant protein was purchased from Sino Biological (100 μg, 40588-V08B) The protein was difficult to dissolve and required treatment before S-trap preparation below with 40 μL dimethyl sulfoxide (276855—100 mL, Sigma), 126 μL 1% TFA, and 100 μL 1× S-Trap lysis buffer (5% SDS, 50 mM TEAB, pH adjusted to 7.55 using 12% phosphoric acid).

    Techniques: Quantitation Assay, Construct

    PRM assay results of mock (SARS-CoV-2 spiked) samples. Three biological replicates processed on different days and averaged from three technical replicates from each mock sample were evaluated using the calibration curves for the two best performing peptides (A) DQVILLNK and (B) FQTLLALHR. The samples represent the spiked-in amounts; low (3.125 μL) and high (12.5 μL) of inactivated SARS-CoV-2 virions into in vitro derived mucus. Tables below display the average calculated amol amounts obtained on each day along with the interday mean and % CV. The dotted line indicates the calculated LOD and the dashed line indicated the LOQ determined from the calibration curves generated for each peptide.

    Journal: Analytical Chemistry

    Article Title: Development of a Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of SARS-CoV-2 Spike Glycoprotein and Nucleoprotein

    doi: 10.1021/acs.analchem.0c02288

    Figure Lengend Snippet: PRM assay results of mock (SARS-CoV-2 spiked) samples. Three biological replicates processed on different days and averaged from three technical replicates from each mock sample were evaluated using the calibration curves for the two best performing peptides (A) DQVILLNK and (B) FQTLLALHR. The samples represent the spiked-in amounts; low (3.125 μL) and high (12.5 μL) of inactivated SARS-CoV-2 virions into in vitro derived mucus. Tables below display the average calculated amol amounts obtained on each day along with the interday mean and % CV. The dotted line indicates the calculated LOD and the dashed line indicated the LOQ determined from the calibration curves generated for each peptide.

    Article Snippet: SARS-CoV-2 nucleocapsid-His recombinant protein was purchased from Sino Biological (100 μg, 40588-V08B) The protein was difficult to dissolve and required treatment before S-trap preparation below with 40 μL dimethyl sulfoxide (276855—100 mL, Sigma), 126 μL 1% TFA, and 100 μL 1× S-Trap lysis buffer (5% SDS, 50 mM TEAB, pH adjusted to 7.55 using 12% phosphoric acid).

    Techniques: In Vitro, Derivative Assay, Generated

    Schematic of the workflow used to develop a PRM assay for the detection and quantitation of SARS-CoV-2 spike and NP (A) PRM assay development was performed using recombinant SARS CoV-2 spike protein and NP. Proteotypic target peptides/transitions were selected to generate a spectral library in Skyline. (B) PRM assay was then used to quantitate the SARS-CoV-2 protein levels in a mock sample that was created by adding an inactivated virus sample to in vitro derived mucus.

    Journal: Analytical Chemistry

    Article Title: Development of a Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of SARS-CoV-2 Spike Glycoprotein and Nucleoprotein

    doi: 10.1021/acs.analchem.0c02288

    Figure Lengend Snippet: Schematic of the workflow used to develop a PRM assay for the detection and quantitation of SARS-CoV-2 spike and NP (A) PRM assay development was performed using recombinant SARS CoV-2 spike protein and NP. Proteotypic target peptides/transitions were selected to generate a spectral library in Skyline. (B) PRM assay was then used to quantitate the SARS-CoV-2 protein levels in a mock sample that was created by adding an inactivated virus sample to in vitro derived mucus.

    Article Snippet: SARS-CoV-2 nucleocapsid-His recombinant protein was purchased from Sino Biological (100 μg, 40588-V08B) The protein was difficult to dissolve and required treatment before S-trap preparation below with 40 μL dimethyl sulfoxide (276855—100 mL, Sigma), 126 μL 1% TFA, and 100 μL 1× S-Trap lysis buffer (5% SDS, 50 mM TEAB, pH adjusted to 7.55 using 12% phosphoric acid).

    Techniques: Quantitation Assay, Recombinant, In Vitro, Derivative Assay

    HKU1 antibodies are prevalent in healthy children and children with acute COVID-19 and MIS-C. SARS-CoV-2 (A) and HKU1 (B) spike IgG antibody titers and FRNT neutralization titers (C) in healthy pediatric controls compared to children hospitalized with acute COVID-19 and MIS-C. * P

    Journal: medRxiv

    Article Title: Original antigenic sin responses to heterologous Betacoronavirus spike proteins are observed in mice following intramuscular administration, but are not apparent in children following SARS-CoV-2 infection

    doi: 10.1101/2021.04.29.21256344

    Figure Lengend Snippet: HKU1 antibodies are prevalent in healthy children and children with acute COVID-19 and MIS-C. SARS-CoV-2 (A) and HKU1 (B) spike IgG antibody titers and FRNT neutralization titers (C) in healthy pediatric controls compared to children hospitalized with acute COVID-19 and MIS-C. * P

    Article Snippet: One group of five Balb/c mice was primed and boosted at 21 days IM with 10 µg SARS-CoV-2 nucleocapsid protein (SinoBiological, 40588-V08B) in 50 µl with alum (Alhydrogel adjuvant 2%, Invivogen).

    Techniques: Neutralization

    Schematic of intramuscular spike protein administrations in groups of five BALB/c mice. Group 1 received prime and boost with SARS-CoV-2 spike, followed by prime and boost with HKU1 spike. Group 2 received a reciprocal administration regimen, with prime and boost with HKU1 spike, followed by prime and boost by HKU1 spike. D, days post-administration; S, spike; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2. *These mice were immunized with nucleocapsid protein 21 and 42 days prior to utilization for this study.

    Journal: medRxiv

    Article Title: Original antigenic sin responses to heterologous Betacoronavirus spike proteins are observed in mice following intramuscular administration, but are not apparent in children following SARS-CoV-2 infection

    doi: 10.1101/2021.04.29.21256344

    Figure Lengend Snippet: Schematic of intramuscular spike protein administrations in groups of five BALB/c mice. Group 1 received prime and boost with SARS-CoV-2 spike, followed by prime and boost with HKU1 spike. Group 2 received a reciprocal administration regimen, with prime and boost with HKU1 spike, followed by prime and boost by HKU1 spike. D, days post-administration; S, spike; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2. *These mice were immunized with nucleocapsid protein 21 and 42 days prior to utilization for this study.

    Article Snippet: One group of five Balb/c mice was primed and boosted at 21 days IM with 10 µg SARS-CoV-2 nucleocapsid protein (SinoBiological, 40588-V08B) in 50 µl with alum (Alhydrogel adjuvant 2%, Invivogen).

    Techniques: Mouse Assay

    HKU1 spike IgG antibodies correlated positively with both SAR-CoV-2 spike IgG and SARS-CoV-2 neutralizing antibodies in children with acute COVID-19 and MIS-C. Linear regression analyses compared the log-transformed antibody titers of (A) SARS-CoV-2 spike IgG vs. HKU1 spike IgG; (B) HKU1 spike IgG vs. SARS-CoV-2 neutralization titers; and (C) SARS-CoV-2 spike IgG vs. SARS-CoV-2 neutralization titers among children with acute COVID-19 or MIS-C. Spearman’s correlation coefficients (r) and P-values are shown.

    Journal: medRxiv

    Article Title: Original antigenic sin responses to heterologous Betacoronavirus spike proteins are observed in mice following intramuscular administration, but are not apparent in children following SARS-CoV-2 infection

    doi: 10.1101/2021.04.29.21256344

    Figure Lengend Snippet: HKU1 spike IgG antibodies correlated positively with both SAR-CoV-2 spike IgG and SARS-CoV-2 neutralizing antibodies in children with acute COVID-19 and MIS-C. Linear regression analyses compared the log-transformed antibody titers of (A) SARS-CoV-2 spike IgG vs. HKU1 spike IgG; (B) HKU1 spike IgG vs. SARS-CoV-2 neutralization titers; and (C) SARS-CoV-2 spike IgG vs. SARS-CoV-2 neutralization titers among children with acute COVID-19 or MIS-C. Spearman’s correlation coefficients (r) and P-values are shown.

    Article Snippet: One group of five Balb/c mice was primed and boosted at 21 days IM with 10 µg SARS-CoV-2 nucleocapsid protein (SinoBiological, 40588-V08B) in 50 µl with alum (Alhydrogel adjuvant 2%, Invivogen).

    Techniques: Transformation Assay, Neutralization

    Priming mice with HKU1 spike protein prior to boosting with SARS-CoV-2 spike protein completely impeded the development of SARS-CoV-2 neutralizing antibodies. SARS-CoV-2 (A,B) and HKU1 (C,D) full-length spike IgG binding and SARS-CoV-2 neutralizing (E, F) antibodies in mice are shown as log(end-point titer). Group 1 was primed with two doses of alum-adjuvanted SARS-CoV-2 spike and boosted with two doses of alum-adjuvanted HKU1 spike (A, C, E). Group 2 received the reciprocal regimen of HKU1 spike prime and SARS-CoV-2 spike boost (B, D, F). * P

    Journal: medRxiv

    Article Title: Original antigenic sin responses to heterologous Betacoronavirus spike proteins are observed in mice following intramuscular administration, but are not apparent in children following SARS-CoV-2 infection

    doi: 10.1101/2021.04.29.21256344

    Figure Lengend Snippet: Priming mice with HKU1 spike protein prior to boosting with SARS-CoV-2 spike protein completely impeded the development of SARS-CoV-2 neutralizing antibodies. SARS-CoV-2 (A,B) and HKU1 (C,D) full-length spike IgG binding and SARS-CoV-2 neutralizing (E, F) antibodies in mice are shown as log(end-point titer). Group 1 was primed with two doses of alum-adjuvanted SARS-CoV-2 spike and boosted with two doses of alum-adjuvanted HKU1 spike (A, C, E). Group 2 received the reciprocal regimen of HKU1 spike prime and SARS-CoV-2 spike boost (B, D, F). * P

    Article Snippet: One group of five Balb/c mice was primed and boosted at 21 days IM with 10 µg SARS-CoV-2 nucleocapsid protein (SinoBiological, 40588-V08B) in 50 µl with alum (Alhydrogel adjuvant 2%, Invivogen).

    Techniques: Mouse Assay, Binding Assay