Protein engineering with artificial chemical nucleasesElectronic supplementary information (ESI) available: Experimental procedures and biological evaluation studies. CCDC 1403896. For ESI and crystallographic data in CIF or other electronic format see DOI: 10.1039/c5cc04615g

Herein we report the application of oxidative artificial chemical nucleases as novel agents for protein engineering. The complex ion [Cu(Phen) 2 (H 2 O)] 2+ (CuPhen; Phen = 1,10-phenanthroline) was applied under Fenton-type conditions against a recombinant antibody fragment specific for prostate-specific antigen (PSA) and compared against traditional DNA shuffling using DNase I for the generation of recombinant mutagenesis libraries. We show that digestion and re-annealment of single chain variable fragment (scFv) coding DNA is possible using CuPhen. Results indicate recombinant library generation in this manner may generate novel clones-not accessible through the use of DNase I-with CuPhen producing highly PSA-specific binding antibodies identified by surface plasmon resonance. The process of protein engineering using artificial chemical nucleases is reported using the Cu( ii )-bis-1,10-phenanthroline complex.