Fig. 3: Characterisation of CD83 scFv and rH9HA-CD83 scFv for binding to chicken CD83 ectodomain using ELISA.

a Indirect ELISA for testing the activity of CD83 mAb b Indirect ELISA for testing the activity of CD83 scFv and rH9HA-CD83 scFv. Purified 8 μg of chicken CD83 ectodomain was coated onto each well of the ELISA plate, a two-fold serial dilution was carried out and the plate was incubated overnight for 4 °C. For detection, the plates were incubated with an equimolar concentration of purified CD83 scFv and rH9HA-CD83 scFv or 1 μg/ml of CD83 mAb. This was followed by incubation with goat anti-mouse HRP secondary antibody for (a) and HRP-conjugated anti-V5 secondary antibody for (b). The colorimetric detection was carried out by adding TMB substrate and absorbance at 450 nm was recorded.