Method from our lab that follow this steps
- Generation of 1000 candidates via Boltzgen
- Selection of first 30 best candidate binders
- ranking accordingly to the largest Buried Solvent Accessibility upon binding (DASA)
- Performing mutagenesis using BeAtMuSiC. Here we use the symmetric version
- Selecting the mutation that increase the binding affinity more than -0.6
- Insert all mutations independently on possible epistatic effects