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

Autor:	Lown PS; Department of Chemical Engineering and Materials Science, University of Minnesota-Twin Cities, Minneapolis, MN 55455, USA., Cai JJ; Department of Chemical Engineering and Materials Science, University of Minnesota-Twin Cities, Minneapolis, MN 55455, USA., Ritter SC; Department of Chemical Engineering and Materials Science, University of Minnesota-Twin Cities, Minneapolis, MN 55455, USA., Otolski JJ; Department of Chemical Engineering and Materials Science, University of Minnesota-Twin Cities, Minneapolis, MN 55455, USA., Wong R; Department of Chemical Engineering and Materials Science, University of Minnesota-Twin Cities, Minneapolis, MN 55455, USA., Hackel BJ; Department of Chemical Engineering and Materials Science, University of Minnesota-Twin Cities, Minneapolis, MN 55455, USA.
Jazyk:	angličtina
Zdroj:	Protein engineering, design & selection : PEDS [Protein Eng Des Sel] 2021 Feb 15; Vol. 34.
DOI:	10.1093/protein/gzab004
Abstrakt:	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. (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
Databáze:	MEDLINE
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