| Flow cytometric screening of yeast surface display libraries. | |
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MedLine Citation:
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PMID: 14976374 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A method to screen and isolate antigen specific clones from a library of single-chain antibodies expressed on the surface of yeast cells is presented. Two rounds of magnetic bead enrichment before flow cytometric sorting enables one to screen libraries of far greater diversity than can be screened by just flow cytometry. The strength of flow cytometric sorting is the ability to follow the selection in real time and to isolate easily the highest affinity antigen-specific clones. A major strength of yeast display as a discovery platform is the ability to characterize the binding properties, the affinity of a clone without the need for subcloning, expression, and purification of the scFv. The methodology for directed evolution of single-chain antibodies to increase the affinity of a clone is also described. |
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Authors:
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Michael Feldhaus; Robert Siegel |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Methods in molecular biology (Clifton, N.J.) Volume: 263 ISSN: 1064-3745 ISO Abbreviation: Methods Mol. Biol. Publication Date: 2004 |
Date Detail:
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Created Date: 2004-02-20 Completed Date: 2004-07-02 Revised Date: 2006-05-01 |
Medline Journal Info:
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Nlm Unique ID: 9214969 Medline TA: Methods Mol Biol Country: United States |
Other Details:
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Languages: eng Pagination: 311-32 Citation Subset: IM |
Affiliation:
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Battelle/Pacific Northwest National Laboratory, Richland, WA, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Agar
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chemistry Antigens / chemistry Biotinylation Culture Media / metabolism Epitopes / chemistry Flow Cytometry / methods* Gene Library* Genes, Fungal* Immunoglobulin Fragments* Kinetics Plasmids / metabolism Saccharomyces cerevisiae / metabolism Time Factors |
| Chemical | |
Reg. No./Substance:
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0/Antigens; 0/Culture Media; 0/Epitopes; 0/Immunoglobulin Fragments; 9002-18-0/Agar |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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