De novo protein design

A) Protein design begins by laying out a backbone that has  the desired shape.
B) Rosetta design is used to identify amino acid sequences with low Rosetta score, good packing, and accurate secondary structure predictions.
C) Designs which have much lower energy than any other conformations sampled in a de-novo folding trajectory are selected.
D) From the best folding designs we order DNA, grow E. Coli transformed with the DNA, and collect our proteins.
E) After purification the quality of the protein design is assessed using CD melts(GuHCl and temperature), SEC-MALS, Mass-spec, SAXS and  Crystallography. Shown is a successful SAXS measurement