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A switchable yeast display/secretion system.

ABSTRACT: 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.

SUBMITTER: Van Deventer JA

PROVIDER: S-EPMC4596280 | biostudies-literature | 2015 Oct

REPOSITORIES: biostudies-literature

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A switchable yeast display/secretion system.

Van Deventer James A JA Kelly Ryan L RL Rajan Saravanan S Wittrup K Dane KD Sidhu Sachdev S SS

Protein engineering, design & selection : PEDS 20150901 10

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, a ...[more]

PMID: 26333274

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