Rapid Affinity Maturation of Novel Anti-PD-L1 Antibodies by a Fast Drop of the Antigen Concentration and FACS Selection of Yeast Libraries

Figure 1

High-throughput sequencing analysis of the CDR3 fragments obtained by random mutagenesis. (a) The chart shows the relative representation of wild type and mutated CDR3 sequences in the indicated cycles of mutagenesis. To estimate the efficiency of mutagenesis, the fraction of fragments showing the nucleotide wild type sequence was evaluated (dotted black line). The amino acid sequences of all the CDR3 fragments were analyzed too, to distinguish between CDR3s showing the wild type sequence (green line-rhomboids) and those containing missense substitutions (blue line-circles) or stop codon triplets (red line-triangles). (b) The histogram represents the distribution of single and multiple mutations in the pools of CDR3s throughout the mutagenesis. The translated sequences were grouped according to the number of amino acid substitutions (indicated in the legend) and are shown as percentage of the mutated sequences.

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