Recombinant Mouse Antibody (E16) is capable of binding to WNV Envelope protein DIII, expressed in Chinese Hamster Ovary cells (CHO).
Figure 1 Mechanism of E16-mediated neutralization of West Nile virus.
a, Two DI/DII-specific neutralizing monoclonal antibodies (E53 and E60) block cellular attachment significantly more than the DIII-specific neutralizing antibodies (E16 and E24) or controls (no antibody, nonneutralizing monoclonal antibody E22 or anti-SARS ORF7a). Foldreductions are reported, with standard deviations, as the average of four to seven independent experiments performed in triplicate. b, Dose-dependent blockade of West Nile virus infection by E16 and E53 in pre- and postadsorption assays. The data are one of three representative experiments performed in duplicate. c, The DIII-specific monoclonal antibodies effectively inhibit West Nile virus infection of macrophages, whereas DI/DII-specific E5 and E60 monoclonal antibodies enhance infection. The data are one of three representative experiments performed in duplicate, with the dotted line representing the limit of sensitivity of the assay. Error bars represent the standard deviation. d, Pre-incubation with unlabelled monoclonal antibodies followed by addition of APC conjugates reveals that both E16 and E60 are self-competitive but not cross-competitive for envelope glycoprotein binding.
Nybakken, G. E., Oliphant, T., Johnson, S., Burke, S., Diamond, M. S., & Fremont, D. H. (2005). Structural basis of West Nile virus neutralization by a therapeutic antibody. Nature, 437(7059), 764.
Figure 2 Surface display of WNV E protein on yeast.
A fusion protein is composed of the ectodomain or DIII of WNV E protein and the yeast Aga2 protein, which becomes attached to the Aga1 protein on the yeast cell wall. Yeast were transformed with the vector alone (pYD1; top row), the entire WNV E ectodomain (amino acids 1–415; middle row), or DIII alone (amino acids 296–415; bottom row). 24 h after induction, yeast were stained with the indicated monoclonal antibodies (negative control, anti-SARS ORF7a) and processed by flow cytometry. Data for a representative neutralizing (E16) and non-neutralizing (E18) antibody are shown.
Oliphant, T., Engle, M., Nybakken, G. E., Doane, C., Johnson, S., Huang, L., ... & Ebel, G. D. (2005). Development of a humanized monoclonal antibody with therapeutic potential against West Nile virus. Nature medicine, 11(5), 522.
Figure 3 Efficacy of mouse E16 in complement and Fcgr1- and Fcgr3-deficient mice.
Dose-response analysis at day 2 after WNV infection. (a) Fcgr1- and Fcgr3-deficient, (b) C1qa-deficient, or (c) C4-deficient 8-week-old C57BL/6J mice were inoculated with 102 PFU of WNV. At 2 d after infection, mice were passively transferred a single dose (4, 20, 100 or 500 µg) of mouse E16. As controls, mice were independently administered saline (PBS). The number of animals for each antibody dose ranged from 10 to 15. The difference in survival curves was statistically significant for all WNV-specific monoclonal antibody doses shown in the C1qa- and C4-deficient mice (P ≤ 0.001). The difference in survival curves for the Fcgr1- and Fcgr3-deficient mice was significant for 500 µg (P = 0.01), but not for 100 µg (P = 0.14) or 20 µg (P = 0.4) of mouse E16.
Oliphant, T., Engle, M., Nybakken, G. E., Doane, C., Johnson, S., Huang, L., ... & Ebel, G. D. (2005). Development of a humanized monoclonal antibody with therapeutic potential against West Nile virus. Nature medicine, 11(5), 522.
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CAT | Product Name | Application | Type |
---|---|---|---|
MOB-1051 | Recombinant Anti-WNV-DIII Antibody | ELISA, WB, FuncS | IgG |
MHH-1051 | Recombinant Human Anti-WNV-DIII Antibody | Neut, WB, FuncS | IgG |
CAT | Product Name | Application | Type |
---|---|---|---|
MOB-1051-F(E) | Recombinant Anti-WNV-DIII Antibody Fab Fragment | FC, IP, FuncS | Fab |
MHH-1051-F(E) | Recombinant Human Anti-WNV-DIII Antibody Fab Fragment | RIA, IF, FuncS | Fab |
CAT | Product Name | Application | Type |
---|---|---|---|
MOB-1051-S(P) | Recombinant Anti-WNV-DIII Antibody scFv Fragment | IF, FC, Biosensors, FuncS | scFv |
MHH-1051-S(P) | Recombinant Human Anti-WNV-DIII Antibody scFv Fragment | WB, IF, FuncS | scFv |
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