Protein engineering with artificial chemical nucleases
| Autor: | Ruth Larragy, Andreea Prisecaru, Andrew Kellett, Jenny Fitzgerald, Vickie McKee, Paul Leonard |
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| Rok vydání: | 2015 |
| Předmět: |
Models
Molecular Iron Molecular Sequence Data Protein Engineering Catalysis law.invention chemistry.chemical_compound Antigen Peptide Library law Materials Chemistry Deoxyribonuclease I Humans Single-chain variable fragment Amino Acid Sequence Surface plasmon resonance Mutagenesis Metals and Alloys DNA Hydrogen Peroxide General Chemistry Protein engineering Prostate-Specific Antigen Molecular biology Recombinant Proteins Surfaces Coatings and Films Electronic Optical and Magnetic Materials DNA shuffling chemistry Biochemistry Ceramics and Composites Recombinant DNA Copper Phenanthrolines Single-Chain Antibodies |
| Zdroj: | Chemical Communications. 51:12908-12911 |
| ISSN: | 1364-548X 1359-7345 |
| DOI: | 10.1039/c5cc04615g |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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