The effect of PSSM-based design restrictions on deimmunization of human erythropoietin (PDB ID 1EER). Designs (n=1000) were generated with (red) and without (blue) an mhc_epitope weight of 1.0 turned on after restricting designable residue identities to those with a base 2 log-odds score of 1, 2, or 3, or with no constraints (NC). The green dashed line indicates the wild-type value (relaxed, but with no design). The effect on six metrics is evaluated: (A) REF2015 score, (B) unweighted mhc_epitope score, (C) PackStat score, and the number of (D) buried unsatisfied hydrogen bonds, (E) positively charged residues, and (F) negatively charged residues. See also Figure S5 and Figure S6 for corresponding plots for other protein targets.