Peer-Reviewed Journal Details
Mandatory Fields
Larragy, R;Fitzgerald, J;Prisecaru, A;Mckee, V;Leonard, P;Kellett, A
2015
January
Chemical Communications
Protein engineering with artificial chemical nucleases
Published
()
Optional Fields
SINGLE-CHAIN FV 1,10-PHENANTHROLINE COPPER BINDING
51
12908
12911
Herein we report the application of oxidative artificial chemical nucleases as novel agents for protein engineering. The complex ion [Cu(Phen)(2)(H2O)](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.
CAMBRIDGE
1359-7345
10.1039/c5cc04615g
Grant Details