期刊
CHEMBIOCHEM
卷 15, 期 12, 页码 1777-1781出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201402184
关键词
antibodies; click chemistry; directed evolution; high-throughput screening; non-canonical amino acids
资金
- NIH [R01 GM62523]
- National Defense Science and Engineering Graduate (NDSEG) fellowship
- National Science Foundation (NSF) Graduate Fellowship
Non-canonical amino acids (ncAAs) provide powerful tools for engineering the chemical and physical properties of proteins. However, introducing ncAAs into proteins can affect protein properties in unpredictable ways, thus necessitating screening efforts to identify mutants with desirable properties. In this work, we describe an Escherichia coli cell surface display platform for the directed evolution of clickable antibody fragments. This platform enabled isolation of antibody fragments with improved digoxigenin binding and modest affinity maturation in several different ncAA contexts. Azide-functionalized fragments exhibited improved binding kinetics relative to their methionine counterparts, facile chemical modification through azide-alkyne cycloaddition, and retention of binding properties after modification. The results described here suggest new possibilities for protein engineering, including modulation of molecular recognition events by ncAAs and direct screening of libraries of chemically modified proteins.
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