When performing directed evolution to optimize a protein’s characteristics, such as thermal stability and biodistribution, many of the produced variants will have lost their original function. To avoid screening these useless variants, researchers use Combinatorial Variant Libraries (CVLs) with soft mutagenesis profiles. CVLs are highly diverse variant libraries engineered to have a complete set of desired mutations across defined amino acid positions. CVL engineering avoids the complications that arise with traditional library fabrication methods, such as premature stop codons, skewed amino acid distributions, and incomplete variant representation, to ensure that each variant retains its original function.
Download this application note from Twist Bioscience to learn how researchers used a CVL to optimize the thermal stability, refolding capacity, and biodistribution of a heterodimeric pair of affibodies.
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