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Flow cytometric evaluation of yeast-bacterial cell-cell interactions.


ABSTRACT: Synthetic cell-cell interaction systems can be useful for understanding multicellular communities or for screening binding molecules. We adapt a previously characterized set of synthetic cognate nanobody-antigen pairs to a yeast-bacteria coincubation format and use flow cytometry to evaluate cell-cell interactions mediated by binding between surface-displayed molecules. We further use fluorescence-activated cell sorting to enrich a specific yeast-displayed nanobody within a mixed yeast-display population. Finally, we demonstrate that this system supports the characterization of a therapeutically relevant nanobody-antigen interaction: a previously discovered nanobody that binds to the intimin protein expressed on the surface of enterohemorrhagic Escherichia coli. Overall, our findings indicate that the yeast-bacteria format supports efficient evaluation of ligand-target interactions. With further development, this format may facilitate systematic characterization and high-throughput discovery of bacterial surface-binding molecules.

SUBMITTER: Lei M 

PROVIDER: S-EPMC10072783 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature

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Flow cytometric evaluation of yeast-bacterial cell-cell interactions.

Lei Ming M   Trivedi Vikas D VD   Nair Nikhil U NU   Lee Kyongbum K   Van Deventer James A JA  

Biotechnology and bioengineering 20221103 2


Synthetic cell-cell interaction systems can be useful for understanding multicellular communities or for screening binding molecules. We adapt a previously characterized set of synthetic cognate nanobody-antigen pairs to a yeast-bacteria coincubation format and use flow cytometry to evaluate cell-cell interactions mediated by binding between surface-displayed molecules. We further use fluorescence-activated cell sorting to enrich a specific yeast-displayed nanobody within a mixed yeast-display p  ...[more]

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