Work

Cell Surface Display Yields Evolvable, Clickable Antibody Fragments

Year: 2014

Type: article

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.

Source: ChemBioChem

DOI: 10.1002/cbic.201402184

Authors: James A. Van Deventer, Kai P. Yuet, Tae Hyeon Yoo, David A. Tirrell

Institutions: California Institute of Technology, Koch Institute for Integrative Cancer Research At MIT, Massachusetts Institute of Technology, Ajou University

Language: English

Topic: Monoclonal and Polyclonal Antibodies Research

Subfield: Radiology, Nuclear Medicine and Imaging

Field: Medicine

Domain: Health Sciences

Open Access status: green

Funder: National Institute of General Medical Sciences

Award: New Amino Acids for Protein Engineering