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nanosota eb2 his  (Millipore)

 
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    Millipore nanosota eb2 his
    (A) Binding affinities between His-tagged nanobodies and two versions of EBOV GP proteins (GP-ΔM and GPcl; see for their definitions) were measured by surface plasmon resonance (SPR). N.D., binding not detected. (B) Binding interactions between His-tagged nanobodies and three versions of the EBOV GP proteins (GP-ΔM, GPcl and sGP; see for their definitions) were evaluated by ELISA. A 450 : absorbances at 450 nm. Data are presented as mean ± SEM (n = 3). An unpaired two-tailed Student’s t -test was used to analyze the statistical differences between the indicated groups, with results indicated above each bar. ****P < 0.0001. (C) Efficacy of Fc-tagged nanobodies in neutralizing EBOV pseudoviruses. Retroviruses pseudotyped with full-length EBOV GP were used to infect Huh7 cells in the presence of Fc-tagged Nansota-EB1 or <t>-EB2</t> at different concentrations. The efficacy of each nanobody against EBOV pseudoviruses was expressed as the concentration required to neutralize pseudovirus entry by 50% (IC 50 ). Error bars represent SEM (n = 3). (D) Efficacy of Fc-tagged nanobodies in neutralizing authentic EBOV infection. Authentic EBOV was used to infect Huh7 cells in the presence of Fc-tagged Nanosota-EB1 or -EB2 at different concentrations. The efficacy of each nanobody against authentic EBOV infection was expressed as the concentration required to neutralize EBOV infection by 50% (IC 50 ). Error bars represent SEM (n = 3). Since Nanosota-EB1-Fc could not fully block viral entry at any of the tested concentrations, the IC 50 values for Nanosota-EB1-Fc are estimations.
    Nanosota Eb2 His, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nanosota eb2 his/product/Millipore
    Average 86 stars, based on 1 article reviews
    nanosota eb2 his - by Bioz Stars, 2025-03
    86/100 stars

    Images

    1) Product Images from "Discovery of Nanosota-EB1 and -EB2 as Novel Nanobody Inhibitors Against Ebola Virus Infection"

    Article Title: Discovery of Nanosota-EB1 and -EB2 as Novel Nanobody Inhibitors Against Ebola Virus Infection

    Journal: PLOS Pathogens

    doi: 10.1371/journal.ppat.1012817

    (A) Binding affinities between His-tagged nanobodies and two versions of EBOV GP proteins (GP-ΔM and GPcl; see for their definitions) were measured by surface plasmon resonance (SPR). N.D., binding not detected. (B) Binding interactions between His-tagged nanobodies and three versions of the EBOV GP proteins (GP-ΔM, GPcl and sGP; see for their definitions) were evaluated by ELISA. A 450 : absorbances at 450 nm. Data are presented as mean ± SEM (n = 3). An unpaired two-tailed Student’s t -test was used to analyze the statistical differences between the indicated groups, with results indicated above each bar. ****P < 0.0001. (C) Efficacy of Fc-tagged nanobodies in neutralizing EBOV pseudoviruses. Retroviruses pseudotyped with full-length EBOV GP were used to infect Huh7 cells in the presence of Fc-tagged Nansota-EB1 or -EB2 at different concentrations. The efficacy of each nanobody against EBOV pseudoviruses was expressed as the concentration required to neutralize pseudovirus entry by 50% (IC 50 ). Error bars represent SEM (n = 3). (D) Efficacy of Fc-tagged nanobodies in neutralizing authentic EBOV infection. Authentic EBOV was used to infect Huh7 cells in the presence of Fc-tagged Nanosota-EB1 or -EB2 at different concentrations. The efficacy of each nanobody against authentic EBOV infection was expressed as the concentration required to neutralize EBOV infection by 50% (IC 50 ). Error bars represent SEM (n = 3). Since Nanosota-EB1-Fc could not fully block viral entry at any of the tested concentrations, the IC 50 values for Nanosota-EB1-Fc are estimations.
    Figure Legend Snippet: (A) Binding affinities between His-tagged nanobodies and two versions of EBOV GP proteins (GP-ΔM and GPcl; see for their definitions) were measured by surface plasmon resonance (SPR). N.D., binding not detected. (B) Binding interactions between His-tagged nanobodies and three versions of the EBOV GP proteins (GP-ΔM, GPcl and sGP; see for their definitions) were evaluated by ELISA. A 450 : absorbances at 450 nm. Data are presented as mean ± SEM (n = 3). An unpaired two-tailed Student’s t -test was used to analyze the statistical differences between the indicated groups, with results indicated above each bar. ****P < 0.0001. (C) Efficacy of Fc-tagged nanobodies in neutralizing EBOV pseudoviruses. Retroviruses pseudotyped with full-length EBOV GP were used to infect Huh7 cells in the presence of Fc-tagged Nansota-EB1 or -EB2 at different concentrations. The efficacy of each nanobody against EBOV pseudoviruses was expressed as the concentration required to neutralize pseudovirus entry by 50% (IC 50 ). Error bars represent SEM (n = 3). (D) Efficacy of Fc-tagged nanobodies in neutralizing authentic EBOV infection. Authentic EBOV was used to infect Huh7 cells in the presence of Fc-tagged Nanosota-EB1 or -EB2 at different concentrations. The efficacy of each nanobody against authentic EBOV infection was expressed as the concentration required to neutralize EBOV infection by 50% (IC 50 ). Error bars represent SEM (n = 3). Since Nanosota-EB1-Fc could not fully block viral entry at any of the tested concentrations, the IC 50 values for Nanosota-EB1-Fc are estimations.

    Techniques Used: Binding Assay, SPR Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Concentration Assay, Infection, Blocking Assay

    (A) Cryo-EM structure of EBOV GP-ΔM complexed with Nanosota-EB2 (top view; surface presentation). The three subunits of EBOV GP-ΔM are colored orange, gray, and green, respectively. Nanosota-EB2 is shown in blue. The N563 glycan involved in binding Nanosota-EB2 is shown in red. The trimeric GP-ΔM is bound by three Nanosota-EB2 molecules. (B) Cryo-EM structure of EBOV GP-ΔM complexed with Nanosota-EB2 (side view). The overall structure is shown in surface presentation, with one Nanosota-EB2 molecule and several membrane-fusion elements in GP shown in cartoon presentation, and the N563 glycan shown in sticks. (C) The binding interface between Nanosota-EB2 and GP. Nanosota-EB2 recognizes quaternary epitopes, including HR1, fusion loop, N-terminus of GP2, and β1/β2 strands of GP1. (D)-(H) Detailed interactions between Nanosota-EB2 and N563 glycan, HR1, fusion loop, N-terminus of GP2, and β1/β2 strands of GP1, respectively. Dotted lines indicate hydrogen bonds. Double arrows indicate hydrophobic interactions.
    Figure Legend Snippet: (A) Cryo-EM structure of EBOV GP-ΔM complexed with Nanosota-EB2 (top view; surface presentation). The three subunits of EBOV GP-ΔM are colored orange, gray, and green, respectively. Nanosota-EB2 is shown in blue. The N563 glycan involved in binding Nanosota-EB2 is shown in red. The trimeric GP-ΔM is bound by three Nanosota-EB2 molecules. (B) Cryo-EM structure of EBOV GP-ΔM complexed with Nanosota-EB2 (side view). The overall structure is shown in surface presentation, with one Nanosota-EB2 molecule and several membrane-fusion elements in GP shown in cartoon presentation, and the N563 glycan shown in sticks. (C) The binding interface between Nanosota-EB2 and GP. Nanosota-EB2 recognizes quaternary epitopes, including HR1, fusion loop, N-terminus of GP2, and β1/β2 strands of GP1. (D)-(H) Detailed interactions between Nanosota-EB2 and N563 glycan, HR1, fusion loop, N-terminus of GP2, and β1/β2 strands of GP1, respectively. Dotted lines indicate hydrogen bonds. Double arrows indicate hydrophobic interactions.

    Techniques Used: Cryo-EM Sample Prep, Binding Assay, Membrane

    (A) Differential scanning fluorimetry (DSF) assay for assessing the impact of nanobodies on the thermal stability of EBOV GP-ΔM. His-tagged Nanosota-EB1 and -EB2 slightly and significantly increased the thermostability of EBOV GP-ΔM, respectively. Comparisons of the Tm values for GP-ΔM in the absence or presence of the nanobodies were performed using an unpaired two-tailed Student’s t -test. Error bars represent SEM (n = 6). ** p <0.01, **** p <0.0001. (B) DSF assay for assessing the impact of His-tagged Nanosota-EB2 on the thermal stability of EBOV GPcl at lower pHs. Nanosota-EB2 significantly increased the thermostability of EBOV GPcl at low pH. Comparisons of the Tm values for GPcl in the absence or presence of Nanosota-EB2 were performed using an unpaired two-tailed Student’s t -test. Error bars represent SEM (n = 6). **** p <0.0001. n.s.: not significant. Note that this experiment could not be conducted for Nanosota-EB1 because Nanosota-EB1 does not bind to EBOV GPcl. (C) Glycan cap cleavage assay to evaluate the effect of Nanosota-EB1 on the protease sensitivity of the glycan cap, using SDS-PAGE under reducing conditions and Coomassie blue staining. Nanosota-EB1 presence slowed the thermolysin L cleavage of GP-ΔM. (D) Glycan cap cleavage assay to evaluate the effect of Nanosota-EB1 on the protease sensitivity of the glycan cap, using Western blot to detect the His tag on GP-ΔM under non-reducing conditions. Nanosota-EB1 presence again slowed thermolysin L cleavage of GP-ΔM. Each of the above experiments was performed three times, yielding consistent results.
    Figure Legend Snippet: (A) Differential scanning fluorimetry (DSF) assay for assessing the impact of nanobodies on the thermal stability of EBOV GP-ΔM. His-tagged Nanosota-EB1 and -EB2 slightly and significantly increased the thermostability of EBOV GP-ΔM, respectively. Comparisons of the Tm values for GP-ΔM in the absence or presence of the nanobodies were performed using an unpaired two-tailed Student’s t -test. Error bars represent SEM (n = 6). ** p <0.01, **** p <0.0001. (B) DSF assay for assessing the impact of His-tagged Nanosota-EB2 on the thermal stability of EBOV GPcl at lower pHs. Nanosota-EB2 significantly increased the thermostability of EBOV GPcl at low pH. Comparisons of the Tm values for GPcl in the absence or presence of Nanosota-EB2 were performed using an unpaired two-tailed Student’s t -test. Error bars represent SEM (n = 6). **** p <0.0001. n.s.: not significant. Note that this experiment could not be conducted for Nanosota-EB1 because Nanosota-EB1 does not bind to EBOV GPcl. (C) Glycan cap cleavage assay to evaluate the effect of Nanosota-EB1 on the protease sensitivity of the glycan cap, using SDS-PAGE under reducing conditions and Coomassie blue staining. Nanosota-EB1 presence slowed the thermolysin L cleavage of GP-ΔM. (D) Glycan cap cleavage assay to evaluate the effect of Nanosota-EB1 on the protease sensitivity of the glycan cap, using Western blot to detect the His tag on GP-ΔM under non-reducing conditions. Nanosota-EB1 presence again slowed thermolysin L cleavage of GP-ΔM. Each of the above experiments was performed three times, yielding consistent results.

    Techniques Used: Two Tailed Test, Cleavage Assay, SDS Page, Staining, Western Blot

    (A) Footprint of Nanosota-EB1 on the glycan cap of EBOV GP (surface representation). (B) Footprint of Nanosota-EB2 on the quaternary epitopes of EBOV GP (surface representation). Nanobody residues are labeled in blue. Nanobody-contacting residues on EBOV GP are categorized as follows: residues conserved between EBOV and BDBV or SUDV are marked in orange, while those differing between EBOV and BDBV or SUDV are highlighted in pink. (C) ELISA results showing the binding interactions between Nanosota-EB1-Fc and GP-ΔM from EBOV, BDBV, and SUDV. (D) ELISA results showing the binding interactions between Nanosota-EB2-Fc and GP-ΔM from EBOV, BDBV, and SUDV. (E) Neutralization efficacy of Fc-tagged nanobodies against BDBV pseudoviruses. (F) Neutralization efficacy of Fc-tagged nanobodies against SUDV pseudoviruses.
    Figure Legend Snippet: (A) Footprint of Nanosota-EB1 on the glycan cap of EBOV GP (surface representation). (B) Footprint of Nanosota-EB2 on the quaternary epitopes of EBOV GP (surface representation). Nanobody residues are labeled in blue. Nanobody-contacting residues on EBOV GP are categorized as follows: residues conserved between EBOV and BDBV or SUDV are marked in orange, while those differing between EBOV and BDBV or SUDV are highlighted in pink. (C) ELISA results showing the binding interactions between Nanosota-EB1-Fc and GP-ΔM from EBOV, BDBV, and SUDV. (D) ELISA results showing the binding interactions between Nanosota-EB2-Fc and GP-ΔM from EBOV, BDBV, and SUDV. (E) Neutralization efficacy of Fc-tagged nanobodies against BDBV pseudoviruses. (F) Neutralization efficacy of Fc-tagged nanobodies against SUDV pseudoviruses.

    Techniques Used: Labeling, Enzyme-linked Immunosorbent Assay, Binding Assay, Neutralization



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    nanosota eb2 his  (Millipore)
    86
    Millipore nanosota eb2 his
    (A) Binding affinities between His-tagged nanobodies and two versions of EBOV GP proteins (GP-ΔM and GPcl; see for their definitions) were measured by surface plasmon resonance (SPR). N.D., binding not detected. (B) Binding interactions between His-tagged nanobodies and three versions of the EBOV GP proteins (GP-ΔM, GPcl and sGP; see for their definitions) were evaluated by ELISA. A 450 : absorbances at 450 nm. Data are presented as mean ± SEM (n = 3). An unpaired two-tailed Student’s t -test was used to analyze the statistical differences between the indicated groups, with results indicated above each bar. ****P < 0.0001. (C) Efficacy of Fc-tagged nanobodies in neutralizing EBOV pseudoviruses. Retroviruses pseudotyped with full-length EBOV GP were used to infect Huh7 cells in the presence of Fc-tagged Nansota-EB1 or <t>-EB2</t> at different concentrations. The efficacy of each nanobody against EBOV pseudoviruses was expressed as the concentration required to neutralize pseudovirus entry by 50% (IC 50 ). Error bars represent SEM (n = 3). (D) Efficacy of Fc-tagged nanobodies in neutralizing authentic EBOV infection. Authentic EBOV was used to infect Huh7 cells in the presence of Fc-tagged Nanosota-EB1 or -EB2 at different concentrations. The efficacy of each nanobody against authentic EBOV infection was expressed as the concentration required to neutralize EBOV infection by 50% (IC 50 ). Error bars represent SEM (n = 3). Since Nanosota-EB1-Fc could not fully block viral entry at any of the tested concentrations, the IC 50 values for Nanosota-EB1-Fc are estimations.
    Nanosota Eb2 His, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nanosota eb2 his/product/Millipore
    Average 86 stars, based on 1 article reviews
    nanosota eb2 his - by Bioz Stars, 2025-03
    86/100 stars
    		  Buy from Supplier

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    (A) Binding affinities between His-tagged nanobodies and two versions of EBOV GP proteins (GP-ΔM and GPcl; see for their definitions) were measured by surface plasmon resonance (SPR). N.D., binding not detected. (B) Binding interactions between His-tagged nanobodies and three versions of the EBOV GP proteins (GP-ΔM, GPcl and sGP; see for their definitions) were evaluated by ELISA. A 450 : absorbances at 450 nm. Data are presented as mean ± SEM (n = 3). An unpaired two-tailed Student’s t -test was used to analyze the statistical differences between the indicated groups, with results indicated above each bar. ****P < 0.0001. (C) Efficacy of Fc-tagged nanobodies in neutralizing EBOV pseudoviruses. Retroviruses pseudotyped with full-length EBOV GP were used to infect Huh7 cells in the presence of Fc-tagged Nansota-EB1 or -EB2 at different concentrations. The efficacy of each nanobody against EBOV pseudoviruses was expressed as the concentration required to neutralize pseudovirus entry by 50% (IC 50 ). Error bars represent SEM (n = 3). (D) Efficacy of Fc-tagged nanobodies in neutralizing authentic EBOV infection. Authentic EBOV was used to infect Huh7 cells in the presence of Fc-tagged Nanosota-EB1 or -EB2 at different concentrations. The efficacy of each nanobody against authentic EBOV infection was expressed as the concentration required to neutralize EBOV infection by 50% (IC 50 ). Error bars represent SEM (n = 3). Since Nanosota-EB1-Fc could not fully block viral entry at any of the tested concentrations, the IC 50 values for Nanosota-EB1-Fc are estimations.

    Journal: PLOS Pathogens

    Article Title: Discovery of Nanosota-EB1 and -EB2 as Novel Nanobody Inhibitors Against Ebola Virus Infection

    doi: 10.1371/journal.ppat.1012817

    Figure Lengend Snippet: (A) Binding affinities between His-tagged nanobodies and two versions of EBOV GP proteins (GP-ΔM and GPcl; see for their definitions) were measured by surface plasmon resonance (SPR). N.D., binding not detected. (B) Binding interactions between His-tagged nanobodies and three versions of the EBOV GP proteins (GP-ΔM, GPcl and sGP; see for their definitions) were evaluated by ELISA. A 450 : absorbances at 450 nm. Data are presented as mean ± SEM (n = 3). An unpaired two-tailed Student’s t -test was used to analyze the statistical differences between the indicated groups, with results indicated above each bar. ****P < 0.0001. (C) Efficacy of Fc-tagged nanobodies in neutralizing EBOV pseudoviruses. Retroviruses pseudotyped with full-length EBOV GP were used to infect Huh7 cells in the presence of Fc-tagged Nansota-EB1 or -EB2 at different concentrations. The efficacy of each nanobody against EBOV pseudoviruses was expressed as the concentration required to neutralize pseudovirus entry by 50% (IC 50 ). Error bars represent SEM (n = 3). (D) Efficacy of Fc-tagged nanobodies in neutralizing authentic EBOV infection. Authentic EBOV was used to infect Huh7 cells in the presence of Fc-tagged Nanosota-EB1 or -EB2 at different concentrations. The efficacy of each nanobody against authentic EBOV infection was expressed as the concentration required to neutralize EBOV infection by 50% (IC 50 ). Error bars represent SEM (n = 3). Since Nanosota-EB1-Fc could not fully block viral entry at any of the tested concentrations, the IC 50 values for Nanosota-EB1-Fc are estimations.

    Article Snippet: Briefly, 60 μg of EBOV GP-ΔM complexed with either Nanosota-EB1-His or Nanosota-EB2-His (100 μg) was treated with 0.25 μg of thermolysin L (Sigma-Aldrich) at 37°C for varying durations (5, 15, or 30 minutes).

    Techniques: Binding Assay, SPR Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Concentration Assay, Infection, Blocking Assay

    (A) Cryo-EM structure of EBOV GP-ΔM complexed with Nanosota-EB2 (top view; surface presentation). The three subunits of EBOV GP-ΔM are colored orange, gray, and green, respectively. Nanosota-EB2 is shown in blue. The N563 glycan involved in binding Nanosota-EB2 is shown in red. The trimeric GP-ΔM is bound by three Nanosota-EB2 molecules. (B) Cryo-EM structure of EBOV GP-ΔM complexed with Nanosota-EB2 (side view). The overall structure is shown in surface presentation, with one Nanosota-EB2 molecule and several membrane-fusion elements in GP shown in cartoon presentation, and the N563 glycan shown in sticks. (C) The binding interface between Nanosota-EB2 and GP. Nanosota-EB2 recognizes quaternary epitopes, including HR1, fusion loop, N-terminus of GP2, and β1/β2 strands of GP1. (D)-(H) Detailed interactions between Nanosota-EB2 and N563 glycan, HR1, fusion loop, N-terminus of GP2, and β1/β2 strands of GP1, respectively. Dotted lines indicate hydrogen bonds. Double arrows indicate hydrophobic interactions.

    Journal: PLOS Pathogens

    Article Title: Discovery of Nanosota-EB1 and -EB2 as Novel Nanobody Inhibitors Against Ebola Virus Infection

    doi: 10.1371/journal.ppat.1012817

    Figure Lengend Snippet: (A) Cryo-EM structure of EBOV GP-ΔM complexed with Nanosota-EB2 (top view; surface presentation). The three subunits of EBOV GP-ΔM are colored orange, gray, and green, respectively. Nanosota-EB2 is shown in blue. The N563 glycan involved in binding Nanosota-EB2 is shown in red. The trimeric GP-ΔM is bound by three Nanosota-EB2 molecules. (B) Cryo-EM structure of EBOV GP-ΔM complexed with Nanosota-EB2 (side view). The overall structure is shown in surface presentation, with one Nanosota-EB2 molecule and several membrane-fusion elements in GP shown in cartoon presentation, and the N563 glycan shown in sticks. (C) The binding interface between Nanosota-EB2 and GP. Nanosota-EB2 recognizes quaternary epitopes, including HR1, fusion loop, N-terminus of GP2, and β1/β2 strands of GP1. (D)-(H) Detailed interactions between Nanosota-EB2 and N563 glycan, HR1, fusion loop, N-terminus of GP2, and β1/β2 strands of GP1, respectively. Dotted lines indicate hydrogen bonds. Double arrows indicate hydrophobic interactions.

    Article Snippet: Briefly, 60 μg of EBOV GP-ΔM complexed with either Nanosota-EB1-His or Nanosota-EB2-His (100 μg) was treated with 0.25 μg of thermolysin L (Sigma-Aldrich) at 37°C for varying durations (5, 15, or 30 minutes).

    Techniques: Cryo-EM Sample Prep, Binding Assay, Membrane

    (A) Differential scanning fluorimetry (DSF) assay for assessing the impact of nanobodies on the thermal stability of EBOV GP-ΔM. His-tagged Nanosota-EB1 and -EB2 slightly and significantly increased the thermostability of EBOV GP-ΔM, respectively. Comparisons of the Tm values for GP-ΔM in the absence or presence of the nanobodies were performed using an unpaired two-tailed Student’s t -test. Error bars represent SEM (n = 6). ** p <0.01, **** p <0.0001. (B) DSF assay for assessing the impact of His-tagged Nanosota-EB2 on the thermal stability of EBOV GPcl at lower pHs. Nanosota-EB2 significantly increased the thermostability of EBOV GPcl at low pH. Comparisons of the Tm values for GPcl in the absence or presence of Nanosota-EB2 were performed using an unpaired two-tailed Student’s t -test. Error bars represent SEM (n = 6). **** p <0.0001. n.s.: not significant. Note that this experiment could not be conducted for Nanosota-EB1 because Nanosota-EB1 does not bind to EBOV GPcl. (C) Glycan cap cleavage assay to evaluate the effect of Nanosota-EB1 on the protease sensitivity of the glycan cap, using SDS-PAGE under reducing conditions and Coomassie blue staining. Nanosota-EB1 presence slowed the thermolysin L cleavage of GP-ΔM. (D) Glycan cap cleavage assay to evaluate the effect of Nanosota-EB1 on the protease sensitivity of the glycan cap, using Western blot to detect the His tag on GP-ΔM under non-reducing conditions. Nanosota-EB1 presence again slowed thermolysin L cleavage of GP-ΔM. Each of the above experiments was performed three times, yielding consistent results.

    Journal: PLOS Pathogens

    Article Title: Discovery of Nanosota-EB1 and -EB2 as Novel Nanobody Inhibitors Against Ebola Virus Infection

    doi: 10.1371/journal.ppat.1012817

    Figure Lengend Snippet: (A) Differential scanning fluorimetry (DSF) assay for assessing the impact of nanobodies on the thermal stability of EBOV GP-ΔM. His-tagged Nanosota-EB1 and -EB2 slightly and significantly increased the thermostability of EBOV GP-ΔM, respectively. Comparisons of the Tm values for GP-ΔM in the absence or presence of the nanobodies were performed using an unpaired two-tailed Student’s t -test. Error bars represent SEM (n = 6). ** p <0.01, **** p <0.0001. (B) DSF assay for assessing the impact of His-tagged Nanosota-EB2 on the thermal stability of EBOV GPcl at lower pHs. Nanosota-EB2 significantly increased the thermostability of EBOV GPcl at low pH. Comparisons of the Tm values for GPcl in the absence or presence of Nanosota-EB2 were performed using an unpaired two-tailed Student’s t -test. Error bars represent SEM (n = 6). **** p <0.0001. n.s.: not significant. Note that this experiment could not be conducted for Nanosota-EB1 because Nanosota-EB1 does not bind to EBOV GPcl. (C) Glycan cap cleavage assay to evaluate the effect of Nanosota-EB1 on the protease sensitivity of the glycan cap, using SDS-PAGE under reducing conditions and Coomassie blue staining. Nanosota-EB1 presence slowed the thermolysin L cleavage of GP-ΔM. (D) Glycan cap cleavage assay to evaluate the effect of Nanosota-EB1 on the protease sensitivity of the glycan cap, using Western blot to detect the His tag on GP-ΔM under non-reducing conditions. Nanosota-EB1 presence again slowed thermolysin L cleavage of GP-ΔM. Each of the above experiments was performed three times, yielding consistent results.

    Article Snippet: Briefly, 60 μg of EBOV GP-ΔM complexed with either Nanosota-EB1-His or Nanosota-EB2-His (100 μg) was treated with 0.25 μg of thermolysin L (Sigma-Aldrich) at 37°C for varying durations (5, 15, or 30 minutes).

    Techniques: Two Tailed Test, Cleavage Assay, SDS Page, Staining, Western Blot

    (A) Footprint of Nanosota-EB1 on the glycan cap of EBOV GP (surface representation). (B) Footprint of Nanosota-EB2 on the quaternary epitopes of EBOV GP (surface representation). Nanobody residues are labeled in blue. Nanobody-contacting residues on EBOV GP are categorized as follows: residues conserved between EBOV and BDBV or SUDV are marked in orange, while those differing between EBOV and BDBV or SUDV are highlighted in pink. (C) ELISA results showing the binding interactions between Nanosota-EB1-Fc and GP-ΔM from EBOV, BDBV, and SUDV. (D) ELISA results showing the binding interactions between Nanosota-EB2-Fc and GP-ΔM from EBOV, BDBV, and SUDV. (E) Neutralization efficacy of Fc-tagged nanobodies against BDBV pseudoviruses. (F) Neutralization efficacy of Fc-tagged nanobodies against SUDV pseudoviruses.

    Journal: PLOS Pathogens

    Article Title: Discovery of Nanosota-EB1 and -EB2 as Novel Nanobody Inhibitors Against Ebola Virus Infection

    doi: 10.1371/journal.ppat.1012817

    Figure Lengend Snippet: (A) Footprint of Nanosota-EB1 on the glycan cap of EBOV GP (surface representation). (B) Footprint of Nanosota-EB2 on the quaternary epitopes of EBOV GP (surface representation). Nanobody residues are labeled in blue. Nanobody-contacting residues on EBOV GP are categorized as follows: residues conserved between EBOV and BDBV or SUDV are marked in orange, while those differing between EBOV and BDBV or SUDV are highlighted in pink. (C) ELISA results showing the binding interactions between Nanosota-EB1-Fc and GP-ΔM from EBOV, BDBV, and SUDV. (D) ELISA results showing the binding interactions between Nanosota-EB2-Fc and GP-ΔM from EBOV, BDBV, and SUDV. (E) Neutralization efficacy of Fc-tagged nanobodies against BDBV pseudoviruses. (F) Neutralization efficacy of Fc-tagged nanobodies against SUDV pseudoviruses.

    Article Snippet: Briefly, 60 μg of EBOV GP-ΔM complexed with either Nanosota-EB1-His or Nanosota-EB2-His (100 μg) was treated with 0.25 μg of thermolysin L (Sigma-Aldrich) at 37°C for varying durations (5, 15, or 30 minutes).

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