A switchable yeast display/secretion system.

TitleA switchable yeast display/secretion system.
Publication TypeJournal Article
Year of Publication2015
AuthorsVan Deventer, James A., Kelly Ryan L., Rajan Saravanan, K Wittrup Dane, and Sidhu Sachdev S.
JournalProtein Eng Des Sel
Volume28
Issue10
Pagination317-25
Date Published2015 Oct
ISSN1741-0134
KeywordsCell Line, Cloning, Molecular, Humans, Models, Molecular, Peptide Library, Protein Conformation, Protein Engineering, Single-Chain Antibodies, Yeasts
Abstract

<p>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.</p>

DOI10.1093/protein/gzv043
Alternate JournalProtein Eng. Des. Sel.
PubMed ID26333274
PubMed Central IDPMC4596280
Grant ListF32 CA168057 / CA / NCI NIH HHS / United States
T32 GM008334 / GM / NIGMS NIH HHS / United States
F32CA168057 / CA / NCI NIH HHS / United States
T32 GM008334-25 / GM / NIGMS NIH HHS / United States

University of Toronto  UCSF  The University of Chicago  QB3  Chicago Biomedical Consortium