In silico design of deimmunized variants. Deimmunization strategy based on algorithm described in supplemental Figure 1. Deimmunization was applied to the neo-epitope identified in VA generated by introducing neo-sequences during protein engineering. (A) Positions in green indicate a predicted decrease in promiscuity scores for the given amino acid substitution. (B) Amino acid substitutions with a decreased promiscuity score were evaluated for conservation score; substitutions depicted in orange indicate substitutions with a conservation score ≥0.6. (C) The amino acid substitutions with a decrease in promiscuity score at nonconserved regions were evaluated using EVmutation; substitutions with a score ≥−0.25 are shown in blue. (D) The Log2 fold change of percent rank of predicted HLA-DRB1 binding compared with VA for each HLA allele. (E) Surface representation of FVII mutant (PDB ID: 3ELA). The potential mutation sites for deimmunization identified using RID are represented as sticks.