期刊
PROTEIN ENGINEERING DESIGN & SELECTION
卷 28, 期 10, 页码 317-325出版社
OXFORD UNIV PRESS
DOI: 10.1093/protein/gzv043
关键词
amber suppression; antibody screening; phage display; synthetic antibody library; yeast display
资金
- Koch Institute
- Ruth L. Kirschstein National Research Service Award [F32CA168057]
- National Institute of General Medical Sciences Interdepartmental Biotechnology Training Program at the National Institutes of Health [T32 GM008334-25]
Display technologies such as yeast and phage display offer powerful alternatives to traditional immunization-based antibody discovery, but require conversion of displayed proteins into soluble form prior to downstream characterization. Here we utilize amber suppression to implement a yeast-based switchable display/secretion system that enables the immediate production of soluble, antibody-like reagents at the end of screening efforts. Model selections in the switchable format remain efficient, and library screening in the switchable format yields renewable sources of affinity reagents exhibiting nanomolar binding affinities. These results confirm that this system provides a seamless link between display-based screening and the production and evaluation of soluble forms of candidate binding proteins. Switchable display/secretion libraries provide a cloning-free, accessible approach to affinity reagent generation.
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