A switchable yeast display/secretion system.
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.
Koch Institute for Integrative Cancer Research Department of Chemical Engineering James.Van_Deventer@tufts.edu sachdev.sidhu@utoronto.ca.
James A
JA
Van Deventer
Koch Institute for Integrative Cancer Research Department of Biological Engineering, Massachusetts Institute of Technology, 500 Main Street, Building 76 Room 289, Cambridge, MA 02139, USA.
Ryan L
RL
Kelly
Department of Molecular Genetics, The Donnelly Centre, University of Toronto, 160 College Street, Toronto, ON, Canada M5S 3E1.
Saravanan
S
Rajan
Koch Institute for Integrative Cancer Research Department of Chemical Engineering Department of Biological Engineering, Massachusetts Institute of Technology, 500 Main Street, Building 76 Room 289, Cambridge, MA 02139, USA.
K Dane
KD
Wittrup
Department of Molecular Genetics, The Donnelly Centre, University of Toronto, 160 College Street, Toronto, ON, Canada M5S 3E1 James.Van_Deventer@tufts.edu sachdev.sidhu@utoronto.ca.
Sachdev S
SS
Sidhu
D002460:Cell Line; D003001:Cloning, Molecular; D006801:Humans; D008958:Models, Molecular; D019151:Peptide Library; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000235:genetics; D015003:Yeasts / Q000235:genetics*
D019151:Peptide Library; D057127:Single-Chain Antibodies
amber suppression; antibody screening; phage display; synthetic antibody library; yeast display