Figure 3.

Affinity maturation of phages displaying Fab HCDR3 mutant of anti-MICA antibodies. After random mutagenesis of CDR3 of 3 anti-MICA phage display clones, they were selected against MICA by biopanning of ten rounds. Phage pools (10⁸ pfu) of each round were tested for MICA binding by ELISA compared to the corresponding phages before mutagenesis (10⁸ pfu). VCSM13 (10⁸ pfu) was used as a negative control (NC). Comparative OD values of each round were shown in (a) WW2G8, (b) WW6B7, and (c) WW9B8. Apparently, the OD's of selections after 7th round were not much increased. The final OD values were increased by 3–5-folds compared to the original clones. Thus, the pool phages of the 7th round were used to isolate individual phages with high binding activities indicated by OD values.