Yeast surface display for screening combinatorial polypeptide libraries.
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.
Boder
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Department of Chemical Engineering, University of Illinois, Urbana 61801, USA.
Wittrup
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D000818:Animals; D003001:Cloning, Molecular; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004926:Escherichia coli; D005434:Flow Cytometry; D015870:Gene Expression; D015698:Genomic Library; D007128:Immunoglobulin Fragments / Q000096:biosynthesis* / Q000737:chemistry; D007700:Kinetics; D008322:Mammals; D000072235:Mating Factor; D008561:Membrane Fusion; D016296:Mutagenesis; D010452:Peptide Biosynthesis; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012323:RNA Processing, Post-Transcriptional; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D007128:Immunoglobulin Fragments; D019151:Peptide Library; D010455:Peptides; D011993:Recombinant Fusion Proteins; D000072235:Mating Factor