Figure 6

Cross-reactivity of the anti-VEEV scFv clones and different anti-Alphavirus specific mAbs analyzed by ELISA. Antigens: VEEV strains TC83, 230 and H12/93 were captured by using anti-VEEV mAb VEE-WIS1 (3 μg/mL); Eastern equine encephalitis virus (EEE), Western equine encephalitis virus (WEE) and Chikungunya (CHIK) were captured by using an anti-Alphavirus mAb mix consisting of mAb 3/4, mAb 12/2 and mAb VEE-WIS1 (3 μg/mL); Culture supernatant of non-infected Vero cells was captured once by anti-VEEV mAb VEE-WIS1 (VERO VEEWIS1) or by a mAb mix consisting of mAb 3/4, mAb 12/2 and mAb VEE-WIS1 (VERO mAb mix). A. Staining with biotinylated anti-VEEV mAb 8/6 (1:10000) and streptavidin conjugated with HRP (1:4000). B. Staining with a biotinylated mixture of antibodies consisting of mAb 8/6 (1:10000), mAb VEE-WIS1 (1:10000), mAb 12/2 (1:5000) and mAb 42/2 (1:2000) followed by a streptavidin-HRP (1:4000) incubation. C. Staining with 1 × 10⁹ (cfu) scFv phage per well was followed by an incubation with mAb anti-M13 conjugated with HRP (1:5000). The IIB6 scFv phage was used as negative control. The mean values of two ELISAs from two independent scFv phage productions are shown.