Figure 2.

High-throughput screening of soft mutagenesis phage libraries for affinity-improved anti-BDNF antibodies.

A: A high-throughput single-point periplasmic competition HTRF assay was used to identify clones with potential improvements in binding over the parental chimeric R3bH01 and humanized H01 in an scFv format. Each point (Red: Round 1 clones, Blue: Round 2 clones, Yellow: Round 3 clones) represents a selection output clone. The data is represented as % inhibition of R3bH01-BDNF interaction compared to the negative control (turquoise, unrelated scFv). Humanized H01 (green) and chimeric R3bH01 (black) as scFv are shown for comparison. B: Prioritized mutants were reformatted to IgG₁ and ranked for potency using a functional pERK signaling assay. R3bH01 (green) is highlighted and compared to B30 (blue). Data was plotted as relative response units versus log molar concentration of purified IgG₁.