Figure 2
From: Multivalent pIX phage display selects for distinct and improved antibody properties
Polyclonal phage ELISA, single-clone screening for mAb 679-14-E06-reactivity and single-clone sequences.
(a) Normalized phage pools from R0 and R2 were analyzed for the presence of binders to the target mAb by polyclonal phage ELISA. The results are given as mean ± SD of duplicates. The experiment was repeated independently giving the same results. (b) Single colonies from R2 were randomly picked for small-scale phagemid rescue and analyzed for target reactivity by ELISA (n = 2). All clones were rescued to HV exploiting functional affinity effects to maximize detection sensitivity. Hence, the LV and HV annotations refer to the valence during selection. A signal/background (S/B) ratio ≥3 was scored as specific (dotted line). Supernatant from empty E. coli TOP10F was included as a control. (c) Random colonies were picked for sequencing from R0 and after R2, and the sequence outputs were compared. The clones from R0 originate from the E. coli transformation outputs, i.e., before separation into LV and HV. Percent sequences within each category and the total number of sequences are indicated in the pie charts.
