Extended yeast surface display linkers enhance the enrichment of ligands in direct mammalian cell selections

Patrick S. Lown, Jessy J. Cai, Seth C. Ritter, Jacob J. Otolski, Ryan Wong, Benjamin J. Hackel

Research output: Contribution to journalArticlepeer-review

2 Scopus citations


Selections of yeast-displayed ligands on mammalian cell monolayers benefit from high target expression and nanomolar affinity, which are not always available. Prior work extending the yeast-protein linker from 40 to 80 amino acids improved yield and enrichment but is hypothesized to be below the optimal length, prompting evaluation of an extended amino acid linker. A 641-residue linker provided enhanced enrichment with a 2-nM affinity fibronectin ligand and 105 epidermal growth factor receptors (EGFR) per cell (14 ± 2 vs. 8 ± 1, P = 0.008) and a >600-nM affinity ligand, 106 EGFR per cell system (23 ± 7 vs. 0.8 ± 0.2, P = 0.004). Enhanced enrichment was also observed with a 310-nM affinity affibody ligand and 104 CD276 per cell, suggesting a generalizable benefit to other scaffolds and targets. Spatial modeling of the linker suggests that improved extracellular accessibility of ligand enables the observed enrichment under conditions not previously possible.

Original languageEnglish (US)
Article numbergzab004
JournalProtein Engineering, Design and Selection
StatePublished - Feb 15 2021

Bibliographical note

Publisher Copyright:
© 2021 The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.


  • ligand
  • protein engineering
  • surface display linker
  • yeast display


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