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PubMed · 9181578

Yeast surface display for screening combinatorial polypeptide libraries.

E T Boder·K D Wittrup
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Abstract

Display on the yeast cell wall is well suited for engineering mammalian cell-surface and secreted proteins (e.g., antibodies, receptors, cytokines) that require endoplasmic reticulum-specific post-translational processing for efficient folding and activity. C-terminal fusion to the Aga2p mating adhesion receptor of Saccharomyces cerevisiae has been used for the selection of scFv antibody fragments with threefold decreased antigen dissociation rate from a randomly mutated library. A eukaryotic host should alleviate expression biases present in bacterially propagated combinatorial libraries. Quantitative flow cytometric analysis enables fine discrimination of kinetic parameters for protein binding to soluble ligands.

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AnimalsCloning, MolecularEndoplasmic ReticulumEscherichia coliFlow CytometryGene ExpressionGenomic LibraryImmunoglobulin FragmentsKineticsMammalsMating FactorMembrane FusionMutagenesisPeptide BiosynthesisPeptide LibraryPeptidesPlasmidsPolymerase Chain ReactionRNA Processing, Post-TranscriptionalRecombinant Fusion ProteinsSaccharomyces cerevisiaeJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, Non-P.H.S.

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E T Boder, K D Wittrup. 1997. Yeast surface display for screening combinatorial polypeptide libraries.. https://doi.org/10.1038/nbt0697-553

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