Fig. 5
Competitive ELISA for characterization of the scFv binding to EBOV VP35. The binding of the scFv-expressing phages (5 × 109 tu/ml) was measured, both in the absence and in the presence of the competitor soluble non-phage-fused scFvs at the maximum concentration of 500 μg/ml. An anti -M13 PVIII coat protein mAb conjugated to HRP was used for phage detection. On the x-axis the concentrations of scFvs used for the competition assay are reported, on the y-axis, the OD450 phage ELISA values are reported. a Competitive ELISA using the anti-GO scFv-expressing phages. The binding of anti-GO scFv-expressing phages to plate coated with GO protein is shown: in the absence of competitor soluble non-phage-fused scFvs; in the presence of their own anti-GO soluble non-phage-fused scFvs at the concentration of 500 μg/ml or 25 μg/ml; in the presence of the anti-VP35 F9, and E1 soluble non-phage-fused scFvs, at the concentration of 500 μg/ml. The higher concentration of anti-GO soluble non-phage-fused scFvs shows to compete with itself. The mean values and the SD of an experiment performed in triplicate are reported. Binding in the presence of soluble non-phage-fused scFvs was compared to binding in presence of the control soluble non-phage-fused scFvs or in their absence using Student’s t-test. *p < 0.05; b Competitive ELISA using anti-EBOV VP35 scFv E1 expressing-phages. The binding of E1 anti-VP35 scFv-expressing phages to plate coated with VP35 protein is shown: in the absence of competitor soluble non-phage-fused scFvs; in the presence of their own E1 soluble non-phage-fused scFvs at the concentration of 500 μg /ml; in the presence of F9 soluble non-phage-fused scFvs at the concentration of 500 μg/ml; in the presence of the control anti-GO soluble non-phage-fused scFvs at the concentration of 500 μg/ml. The mean values and the SD of an experiment performed in triplicate are reported. Binding in the presence of soluble non-phage-fused scFvs was compared to binding in the presence of the control soluble non-phage-fused scFvs or in their absence using Student’s t-test. *p < 0.05