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: Plant Biotechnology Journal

Article Title: Plant‐made dengue virus‐like particles produced by co‐expression of structural and non‐structural proteins induce a humoral immune response in mice

doi: 10.1111/pbi.13501

Figure Lengend Snippet: Analysed candidate vaccines and Coomassie‐stained SDS‐PAGE control: The respective vaccine candidates were expressed in plants, purified and analysed by SDS‐PAGE. DENV1‐VLPs were produced by co‐expressing DENV1‐SP and NSP. SDS‐PAGE shows signals for prM (18 kDa) and E (~55 kDa). Empty bluetongue core‐like particles (BTV‐CLPs) as negative control (c‐) were produced by co‐expression of BTV VP3 (103 kDa) and VP7 (39 kDa). For antigen display inside BTV‐CLPs, the E domain III (EIII) of DENV1, DENV4 and ZIKA was fused to the N terminus of BTV‐VP3. Co‐expression of BTV‐VP3 and V7 leads to assembly of CLPs and integration of the N ‐terminal fused antigens. The Coomassie‐stained SDS‐PAGE shows the size difference between c‐ VP3 and VP3 fused to EIII. The EIII soluble subunit (14 kDa) showed the expected size of 14 kDa and high purity.

Article Snippet: To detect specific antibodies, 96‐well plates (NUNC Maxisorp) were coated with 5 µg/mL of each of the following antigens diluted in carbonate–bicarbonate buffer (Thermo, Product #28382): recombinant Zika virus envelope protein (Native Antigen Company, ZIKVSU‐ENV‐100), Zika VLP (Native Antigen Company, ZIKV‐VLP‐250), DENV1 VLP (Native Antigen Company, DENV1‐VLP‐250) and DENV4 VLP (Native Antigen Company, DENV4‐VLP‐250).

Techniques: Vaccines, Staining, SDS Page, Purification, Produced, Expressing, Negative Control

Journal: Plant Biotechnology Journal

Article Title: Plant‐made dengue virus‐like particles produced by co‐expression of structural and non‐structural proteins induce a humoral immune response in mice

doi: 10.1111/pbi.13501

Figure Lengend Snippet: Humoral response of DENV1 VLPs, EIII domains displayed in bluetongue CLPs and the DENV1‐EIII subunit. Six mice per group were immunized in a single or prime‐boost manner. Collected sera of immunized mice were tested in a dilution series against commercial DENV1, DENV4 and ZIKA VLPs, as well as recombinant ZIKA envelope protein (ZIKV‐Env). Lines show mean values with error bars denoted standard error. Only DENV1‐VLPs stimulated the production of reacting antibodies, indicating that in mice, VLPs are more efficient antigens in terms of B‐cell response than comparable amounts of the DENV1 subunit alone or the display of EIII on bluetongue CLPs.

Article Snippet: To detect specific antibodies, 96‐well plates (NUNC Maxisorp) were coated with 5 µg/mL of each of the following antigens diluted in carbonate–bicarbonate buffer (Thermo, Product #28382): recombinant Zika virus envelope protein (Native Antigen Company, ZIKVSU‐ENV‐100), Zika VLP (Native Antigen Company, ZIKV‐VLP‐250), DENV1 VLP (Native Antigen Company, DENV1‐VLP‐250) and DENV4 VLP (Native Antigen Company, DENV4‐VLP‐250).

Techniques: Recombinant