Fig. 4: Comparing in silico metrics with experimental outcomes for solenoid designs from the in silico evolution platform.

A bar chart showing the outcomes of the experimental characterisation of in silico platform solenoid designs. This is shown by how many designs were evaluated, cloned into a backbone vector, had a purification product at the expected molecular weight after Ni-NTA purification and if there is an expected SEC and CD spectra trace. B–E Metrics are comparing experimental success (S) versus failure (F) in designs, defined by the presence of an expected SEC peak and a matching CD trace for secondary structure. The five successful designs include A_1_1, A_2_1, A_2_2, A_3_1 and A_3_2; the remaining 19 designs are classified as failures. B AF2 quality metrics: Box plots for average pLDDT (left, not significant), average PAE (middle, not significant), and RMSD to the original backbone design post-sequence redesign (right, significant with P-value 0.0093). C ESMFold quality metrics: Box plots for pLDDT (left, not significant), pTM (middle, not significant), and RMSD to the original backbone design post-sequence redesign (right, not significant). D Similarity to natural proteins: Sequence identity with the top hit in the nr database (left, not significant) and TM alignment score to the closest PDB match (right, significant with P-value 0.0044). E Box plots of the sigmoidal inflection point values (representing ‘melting temperatures’) of the ‘melting curves’ (significant with P-value 0.019). Statistical significance was determined using the Mann-Whitney U test.