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. 2016 Feb 23;8(3):574–584. doi: 10.1080/19420862.2016.1148850

Figure 2.

Figure 2.

Bispecific binding capacity of DVD-1A1D-2A10. (A) Western blot analysis of DVD-1A1D-2A10. 1A1D and 2A10 bind DIII and DI/II, respectively; DVD-Ig binds both D I/II and D III. (B) A competition ELISA was used to confirm DVD-1A1D-2A10 binding D I/II and D III simultaneously. (C) Bispecific binding to virus. BHK21 cells were infected with DENV2-43. Three to 5 days after infection, cells were fixed and analyzed by indirect immunofluorescence analysis. First, cells were blocked by murine antibodies (m1A1D, m2A10 and m1A1D+m2A10, respectively). Then, these cells were incubated with corresponding chimeric antibodies (c1A1D, c2A10 and cDVD-1A1D-2A10) and tested by fluorescein isothiocyanate (FITC)–conjugated anti-human IgG. Green means that chimeric antibody could bind to E proteins on the virus surface. Red represent negative cells. (D and E). Binding activity of DVD-1A1D-2A10 compared to parental antibodies. Increasing concentrations of DVD-1A1D-2A10, 1A1D, and 2A10 were added to 96-well plates coated with DI/II (D) or DIII (E). DVD-1A1D-2A10 retained equal binding activity of 1A1D and 2A10, respectively.