Figure 6. Affimer detection of HVT UL49 in infected cells by in-cell Western and affinity-fluorescence.
(A) Phage ELISA for 24 monoclonal Affimers against HVT screened against HVT, RB1B, DEV and CEF lysates to confirm specificity for HVT. (B) Infection of cells was confirmed using a goat anti-GFP antibody and donkey anti-goat 680 (green) antibody to detect GFP which is constitutively expressed by the BAC derived viruses. Infected CEFs were screened with candidate HVT UL49 Affimers at 1.5 µg/ml with subsequent detection (red) by Streptavidin 800 conjugate (Licor). (n = 3) (D) HVT infected CEFs were screened with HVT UL49 Affimers and visualised with Streptavidin-568 (red). Nuclei were stained with DAPI (blue). Streptavidin only control (no Affimer) shows no observable labelling of infected CEFs. Bar = 10 µm.
Figure 6—figure supplement 1. Bilayer Interferometry plots for the HVT binding Affimers.
