Document Detail


Yeast mating for combinatorial Fab library generation and surface display.
MedLine Citation:
PMID:  15094038     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Yeast display of antibody fragments has proven to be an efficient and productive means for directed evolution of single chain Fv antibodies for increased affinity and thermal stability, and more recently for the display and screening of a non-immune library. In this paper, we describe an elegant and simple method for constructing large combinatorial Fab libraries for display on the surface of Saccharomyces cerevisiae, from modestly sized, and easily constructed, heavy and light chain libraries. To this end, we have constructed a set of yeast strains and a two vector system for heavy chain and light chain surface display of Fab fragments with free native amino termini. Through yeast mating of the haploid libraries, a very large heterodimeric immune Fab library was displayed on the diploids and high affinity antigen specific Fabs were isolated from the library.
Authors:
Jane M Weaver-Feldhaus; Jianlong Lou; James R Coleman; Robert W Siegel; James D Marks; Michael J Feldhaus
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  FEBS letters     Volume:  564     ISSN:  0014-5793     ISO Abbreviation:  FEBS Lett.     Publication Date:  2004 Apr 
Date Detail:
Created Date:  2004-04-19     Completed Date:  2004-06-03     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0155157     Medline TA:  FEBS Lett     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  24-34     Citation Subset:  IM    
Affiliation:
Pacific Northwest National Laboratory, MSIN:K4-12, 902 Battelle Boulevard, P.O. Box 999, Richland, WA 99352, USA.
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MeSH Terms
Descriptor/Qualifier:
Antibody Affinity
Antibody Specificity
Combinatorial Chemistry Techniques
Genetic Vectors
Immunoglobulin Fab Fragments / biosynthesis*,  immunology
Peptide Library*
Saccharomyces cerevisiae / genetics
Yeasts / genetics*
Grant Support
ID/Acronym/Agency:
R21 AI53389-01/AI/NIAID NIH HHS; U01 AI056493/AI/NIAID NIH HHS; U54 CA90788/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Immunoglobulin Fab Fragments; 0/Peptide Library

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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