zikv vlps  (Native Antigen Inc)

Journal: Journal of Virology

Article Title: Somatic Hypermutation and Framework Mutations of Variable Region Contribute to Anti-Zika Virus-Specific Monoclonal Antibody Binding and Function

doi: 10.1128/jvi.00071-22

Figure Lengend Snippet: Reactivity of anti-ZIKV MAbs. (A) Reactivity of anti-ZIKV MAbs with ZIKV and DENV virus-like particles (VLP) using Luminex assay. ZIKV- and DENV-VLP were conjugated to MagPlex beads (Luminex) and 10,000 beads/mL of these beads mixed with 10 μg/mL to 0.0002 ng/mL anti-ZIKV MAb 102-1, 242-3, 270-12, 289-3, 306-2, 11-3, 278-11, 78-2, and 4G2 for 90 min at room temperature. (B) Western blot analysis of anti-ZIKV MAbs. ZIKV-VLP were heat denatured under nonreduced conditions at 70°C for 5 min. Samples (left, 27 ng; right, 240 ng) were electrophoresed by Wes capillary and detected by 10 μg/mL anti-ZIKV MAb clones. Left, MAb 102-1, 242-3, 270-12, 289-3, 306-2, 78-2 and 11-3. Right, 278-11. M, molecular weight marker. The estimated molecular mass of ZIKV E protein, 55 kDa; prM protein, 23 kDa.

Article Snippet: DENV-1-VLP (Nauru/Western Pacific/1974), DENV-2-VLP (Thailand/16681/84), DENV-3-VLP (Sri Lanka D3/H/IMTSSA-SRI/2000/1266), DENV-4-VLP (Dominica/814669/1981), ZIKV-VLP (Suriname Z1106033), and ZIKV E protein (Suriname Z1106033) were purchased from The Native Antigen Company (Oxford, UK).

Techniques: Luminex, Western Blot, Clone Assay, Molecular Weight, Marker

Journal: Journal of Virology

Article Title: Somatic Hypermutation and Framework Mutations of Variable Region Contribute to Anti-Zika Virus-Specific Monoclonal Antibody Binding and Function

doi: 10.1128/jvi.00071-22

Figure Lengend Snippet: Summary of characterization of anti-ZIKV MAbs

Article Snippet: DENV-1-VLP (Nauru/Western Pacific/1974), DENV-2-VLP (Thailand/16681/84), DENV-3-VLP (Sri Lanka D3/H/IMTSSA-SRI/2000/1266), DENV-4-VLP (Dominica/814669/1981), ZIKV-VLP (Suriname Z1106033), and ZIKV E protein (Suriname Z1106033) were purchased from The Native Antigen Company (Oxford, UK).

Techniques: Luminex, Western Blot

Journal: Journal of Virology

Article Title: Somatic Hypermutation and Framework Mutations of Variable Region Contribute to Anti-Zika Virus-Specific Monoclonal Antibody Binding and Function

doi: 10.1128/jvi.00071-22

Figure Lengend Snippet: Kinetic analysis of anti-ZIKV MAbs. Kinetic analysis was conducted by Octet HTX (Sartorius). Anti-ZIKV MAbs were conjugated to an amine-reactive 2nd generation (AR2G) biosensor at 0.1 to 0.3 μg/mL, and the association constant ( k a ) was measured over 0 to 900 s and dissociation constant ( k dis ) for 1,200 s for ZIKV-VLP or E proteins at various concentrations. (A) MAb 289-3 and 102-1 to ZIKV-VLP, 3.3 to 16.7 nM. (B) MAb 289-3 and 102-1 to ZIKV E proteins, 6.67 to 33.3 nM, red line, fitting pattern. (C) k a / k dis plot for anti-ZIKV MAbs to ZIKV-VLP and ZIKV E proteins. Blue, MAb 78-2, 289-3, 306-2 and 278-11; red, MAb 102-1, 242-3. 270-12, and 11-3. k a and k dis values were calculated by two runs, and the average values are shown. Values (nM) for equilibrium dissociation constants ( K D ) are shown for each dotted line. K D = K dis / K a .

Article Snippet: DENV-1-VLP (Nauru/Western Pacific/1974), DENV-2-VLP (Thailand/16681/84), DENV-3-VLP (Sri Lanka D3/H/IMTSSA-SRI/2000/1266), DENV-4-VLP (Dominica/814669/1981), ZIKV-VLP (Suriname Z1106033), and ZIKV E protein (Suriname Z1106033) were purchased from The Native Antigen Company (Oxford, UK).

Techniques:

Journal: Journal of Virology

Article Title: Somatic Hypermutation and Framework Mutations of Variable Region Contribute to Anti-Zika Virus-Specific Monoclonal Antibody Binding and Function

doi: 10.1128/jvi.00071-22

Figure Lengend Snippet: Binding activity of framework region (FWR) amino acid reverted anti-ZIKV MAb. (A to D) Association and dissociation analysis of FWR amino acid reverted anti-ZIKV MAbs by Octet HTX (Sartorius); 2 μg/mL MAbs were captured to protein G biosensor (Sartorius), and 3 μg/mL ZIKV E protein was associated for 600 s and dissociated for 900 s. (E to H) Reactivity of anti-ZIKV MAbs of ZIKV-VLP using Luminex assay. Blue, anti-ZIKV MAbs with matured amino acid; red, anti-ZIKV MAbs with amino acid reverted to allele. (A and E) MAb 102-1. (B and F) MAb 270-12. (C and G) MAb 289-3. (D and H) MAb 306-2.

Article Snippet: DENV-1-VLP (Nauru/Western Pacific/1974), DENV-2-VLP (Thailand/16681/84), DENV-3-VLP (Sri Lanka D3/H/IMTSSA-SRI/2000/1266), DENV-4-VLP (Dominica/814669/1981), ZIKV-VLP (Suriname Z1106033), and ZIKV E protein (Suriname Z1106033) were purchased from The Native Antigen Company (Oxford, UK).

Techniques: Binding Assay, Activity Assay, Luminex

Journal: Journal of Virology

Article Title: Somatic Hypermutation and Framework Mutations of Variable Region Contribute to Anti-Zika Virus-Specific Monoclonal Antibody Binding and Function

doi: 10.1128/jvi.00071-22

Figure Lengend Snippet: Reactivity of anti-ZIKV MAbs. (A) Reactivity of anti-ZIKV MAbs with ZIKV and DENV virus-like particles (VLP) using Luminex assay. ZIKV- and DENV-VLP were conjugated to MagPlex beads (Luminex) and 10,000 beads/mL of these beads mixed with 10 μg/mL to 0.0002 ng/mL anti-ZIKV MAb 102-1, 242-3, 270-12, 289-3, 306-2, 11-3, 278-11, 78-2, and 4G2 for 90 min at room temperature. (B) Western blot analysis of anti-ZIKV MAbs. ZIKV-VLP were heat denatured under nonreduced conditions at 70°C for 5 min. Samples (left, 27 ng; right, 240 ng) were electrophoresed by Wes capillary and detected by 10 μg/mL anti-ZIKV MAb clones. Left, MAb 102-1, 242-3, 270-12, 289-3, 306-2, 78-2 and 11-3. Right, 278-11. M, molecular weight marker. The estimated molecular mass of ZIKV E protein, 55 kDa; prM protein, 23 kDa.

Article Snippet: A volume of 0.1 to 1.0 μg/mL ZIKV-VLP or 0.2 to 2 μg/mL ZIKV E protein in 1× kinetic buffer (Sartorius, Fremont, CA, USA) was associated with anti-ZIKV MAb for 900 s and dissociated for 1,200 s. Kinetic parameters, association constant ( k a ), and dissociation constant ( k dis ) were analyzed by Octet Data Analysis Software HT (ver.

Techniques: Virus, Luminex, Western Blot, Clone Assay, Molecular Weight, Marker

Journal: Journal of Virology

Article Title: Somatic Hypermutation and Framework Mutations of Variable Region Contribute to Anti-Zika Virus-Specific Monoclonal Antibody Binding and Function

doi: 10.1128/jvi.00071-22

Figure Lengend Snippet: Summary of characterization of anti-ZIKV MAbs

Article Snippet: A volume of 0.1 to 1.0 μg/mL ZIKV-VLP or 0.2 to 2 μg/mL ZIKV E protein in 1× kinetic buffer (Sartorius, Fremont, CA, USA) was associated with anti-ZIKV MAb for 900 s and dissociated for 1,200 s. Kinetic parameters, association constant ( k a ), and dissociation constant ( k dis ) were analyzed by Octet Data Analysis Software HT (ver.

Techniques: Luminex, Western Blot

Journal: Journal of Virology

Article Title: Somatic Hypermutation and Framework Mutations of Variable Region Contribute to Anti-Zika Virus-Specific Monoclonal Antibody Binding and Function

doi: 10.1128/jvi.00071-22

Figure Lengend Snippet: Kinetic analysis of anti-ZIKV MAbs. Kinetic analysis was conducted by Octet HTX (Sartorius). Anti-ZIKV MAbs were conjugated to an amine-reactive 2nd generation (AR2G) biosensor at 0.1 to 0.3 μg/mL, and the association constant ( k a ) was measured over 0 to 900 s and dissociation constant ( k dis ) for 1,200 s for ZIKV-VLP or E proteins at various concentrations. (A) MAb 289-3 and 102-1 to ZIKV-VLP, 3.3 to 16.7 nM. (B) MAb 289-3 and 102-1 to ZIKV E proteins, 6.67 to 33.3 nM, red line, fitting pattern. (C) k a / k dis plot for anti-ZIKV MAbs to ZIKV-VLP and ZIKV E proteins. Blue, MAb 78-2, 289-3, 306-2 and 278-11; red, MAb 102-1, 242-3. 270-12, and 11-3. k a and k dis values were calculated by two runs, and the average values are shown. Values (nM) for equilibrium dissociation constants ( K D ) are shown for each dotted line. K D = K dis / K a .

Article Snippet: A volume of 0.1 to 1.0 μg/mL ZIKV-VLP or 0.2 to 2 μg/mL ZIKV E protein in 1× kinetic buffer (Sartorius, Fremont, CA, USA) was associated with anti-ZIKV MAb for 900 s and dissociated for 1,200 s. Kinetic parameters, association constant ( k a ), and dissociation constant ( k dis ) were analyzed by Octet Data Analysis Software HT (ver.

Techniques:

Journal: Journal of Virology

Article Title: Somatic Hypermutation and Framework Mutations of Variable Region Contribute to Anti-Zika Virus-Specific Monoclonal Antibody Binding and Function

doi: 10.1128/jvi.00071-22

Figure Lengend Snippet: Binding activity of framework region (FWR) amino acid reverted anti-ZIKV MAb. (A to D) Association and dissociation analysis of FWR amino acid reverted anti-ZIKV MAbs by Octet HTX (Sartorius); 2 μg/mL MAbs were captured to protein G biosensor (Sartorius), and 3 μg/mL ZIKV E protein was associated for 600 s and dissociated for 900 s. (E to H) Reactivity of anti-ZIKV MAbs of ZIKV-VLP using Luminex assay. Blue, anti-ZIKV MAbs with matured amino acid; red, anti-ZIKV MAbs with amino acid reverted to allele. (A and E) MAb 102-1. (B and F) MAb 270-12. (C and G) MAb 289-3. (D and H) MAb 306-2.

Article Snippet: A volume of 0.1 to 1.0 μg/mL ZIKV-VLP or 0.2 to 2 μg/mL ZIKV E protein in 1× kinetic buffer (Sartorius, Fremont, CA, USA) was associated with anti-ZIKV MAb for 900 s and dissociated for 1,200 s. Kinetic parameters, association constant ( k a ), and dissociation constant ( k dis ) were analyzed by Octet Data Analysis Software HT (ver.

Techniques: Binding Assay, Activity Assay, Luminex