Introduction to in Silico and Experimental Validation
Once the sequences have been optimized for the target backbone, it is necessary to test whether these sequences can adopt the target structure. These sequences must be evaluated by computer through protein structure prediction methods before choosing to clear the computer-tested sequences for experimental verification. These sequences will follow the routine procedures for protein expression, purification and structural characterization using biophysical techniques (such as CD, fluorescence). More powerful tests include the structure determination using NMR, X-ray crystallography or even Cryo-EM.