An improved yeast surface display platform for the screening of nanobody immune libraries

Tomasz Uchański, Thomas Zögg, Jie Yin, Daopeng Yuan, Alexandre Wohlkönig, Baptiste Fischer, Daniel M. Rosenbaum, Brian K. Kobilka, Els Pardon, Jan Steyaert*

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

60 Citations (Scopus)

Abstract

Fusions to the C-terminal end of the Aga2p mating adhesion of Saccharomyces cerevisiae have been used in many studies for the selection of affinity reagents by yeast display followed by flow cytometric analysis. Here we present an improved yeast display system for the screening of Nanobody immune libraries where we fused the Nanobody to the N-terminal end of Aga2p to avoid steric hindrance between the fused Nanobody and the antigen. Moreover, the display level of a cloned Nanobody on the surface of an individual yeast cell can be monitored through a covalent fluorophore that is attached in a single enzymatic step to an orthogonal acyl carrier protein (ACP). Additionally, the displayed Nanobody can be easily released from the yeast surface and immobilised on solid surfaces for rapid analysis. To prove the generic nature of this novel Nanobody discovery platform, we conveniently selected Nanobodies against three different antigens, including two membrane proteins.

Original languageEnglish
Article number382
JournalScientific Reports
Volume9
Issue number1
DOIs
Publication statusPublished - 1 Dec 2019
Externally publishedYes

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