Document Detail


Flow cytometric screening of yeast surface display libraries.
MedLine Citation:
PMID:  14976374     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Michael Feldhaus; Robert Siegel
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  263     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2004  
Date Detail:
Created Date:  2004-02-20     Completed Date:  2004-07-02     Revised Date:  2006-05-01    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  311-32     Citation Subset:  IM    
Affiliation:
Battelle/Pacific Northwest National Laboratory, Richland, WA, USA.
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MeSH Terms
Descriptor/Qualifier:
Agar / 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:
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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