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Cell Surface Display Yields Evolvable, Clickable Antibody Fragments
Author(s) -
Van Deventer James A.,
Yuet Kai P.,
Yoo Tae Hyeon,
Tirrell David A.
Publication year - 2014
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201402184
Subject(s) - protein engineering , directed evolution , click chemistry , phage display , chemistry , azide , directed molecular evolution , computational biology , combinatorial chemistry , biochemistry , mutant , biology , enzyme , peptide , organic chemistry , gene
Abstract 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.