zika vlp  (Expression Systems Inc)

Journal: Journal of Virology

Article Title: Virus-like particle vaccine with authentic quaternary epitopes protects against Zika virus-induced viremia and testicular damage

doi: 10.1128/jvi.02322-24

Figure Lengend Snippet: Vaccination of Ifnar1 −/− mice with Zika VLP or SVP and ZIKV challenge. ( A ) Timeline of vaccination of female Ifnar1 −/− mice with a single, non-adjuvanted dose of 1 µg Zika VLP or Zika SVP or CHIK VLP (negative control), followed by antibody measurements, and challenge with ZIKV Natal followed by viremia determinations, or challenge with ZIKV MR766 followed by viremia and survival determinations. ( B ) ZIKV MR766 end point IgG ELISA titers in serum from female Ifnar −/− mice after immunization with one dose of the indicated vaccine. Limit of detection was one in 10 serum dilution. ( C ) ZIKV Natal 50% neutralization titers in serum from female Ifnar1 −/− mice vaccinated as in B. Limit of detection is one in 10 serum dilution. Statistics by Kolmogorov-Smirnov exact tests. (D) Mean viremias post ZIKV challenge ( n = 5 per group). The limit of detection per mouse was 2 log 10 TCID 50 /mL. Statistics relative to CHIK VLP on day 4. ( E ) Survival of immunized mice after ZIKV MR766 challenge. Animals were euthanized when ethically defined end points had been reached. Statistics by log-rank tests relative to CHIK VLP. ( F ) Timeline of vaccination of male Ifnar1 −/− mice with three 1 µg doses of Zika VLP or SVP, or CHIK VLP (negative control) adjuvanted with AddaVax, or PBS (negative control), followed by serum antibody and viremia determinations after challenge with ZIKV PRVABC59 . ( G ) Serum ZIKV MR766 end point IgG ELISA titers after three vaccinations or after infection with ZIKV Natal (positive control). Limit of detection was one in 10 serum dilution. ( H ) Reciprocal anti-ZIKV PRVABC59 50% neutralization titers. Limit of detection was one in 10 serum dilution. ( I ) Mean ZIKV PRVABC59 viremias post challenge ( n = 4–5 per group). Statistics relative to CHIK VLP on day 6 (for Zika VLP) and day 7 (for Zika SVP). Statistics Smirnov exact tests was used for data in panels B, C, D, G, and I.

Article Snippet: Male Ifnar1 −/− mice (≈12 weeks old) were immunized at three different times with 1 µg Zika VLPs or SVPs mixed in a 1:1 vol ratio with AddaVax adjuvant (InvivoGen) ( ).

Techniques: Negative Control, Enzyme-linked Immunosorbent Assay, Neutralization, Infection, Positive Control

Journal: Journal of Virology

Article Title: Virus-like particle vaccine with authentic quaternary epitopes protects against Zika virus-induced viremia and testicular damage

doi: 10.1128/jvi.02322-24

Figure Lengend Snippet: Histopathology of testes after ZIKV PRVABC59 challenge. ( A ) H&E-stained sections of testes from mice that received PBS + adjuvant (negative control). Testes were harvested 31 days after challenge with ZIKV PRVABC59 . ( B ) As for A, but for mice that were vaccinated with SVP-A264C (pH 7) + AddaVax (same mice as described in ). ( C ) Enlargements of A and B showing overt loss of seminiferous tubules or disruption of seminiferous tubule architecture. The SVP-A264C (pH 7) + AddaVax vaccine group shows normal seminiferous tubule morphology. S, lumen of the seminiferous tubules.

Article Snippet: Male Ifnar1 −/− mice (≈12 weeks old) were immunized at three different times with 1 µg Zika VLPs or SVPs mixed in a 1:1 vol ratio with AddaVax adjuvant (InvivoGen) ( ).

Techniques: Histopathology, Staining, Adjuvant, Negative Control, Disruption

Journal:

Article Title:

doi:

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:

Article Title:

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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:

Article Title:

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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).

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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