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Construction and characterization of a pseudo-immune human antibody library using yeast surface display.

Authors
Lee, HW; Lee, SH; Park, KJ; Kim, JS; Kwon, MH; Kim, YS
Citation
Biochemical and biophysical research communications, 346(3):896-903, 2006
Journal Title
Biochemical and biophysical research communications
ISSN
0006-291X1090-2104
Abstract
Lymphocytes from eight individuals out of 60 healthy donors, whose plasmas showed relatively higher antibody titer for a target antigen of death receptor 5 (DR5), were selected for the source of antibody genes to construct so called an anti-DR5 pseudo-immune human single-chain fragment variable (scFv) library on the yeast cell surface (approximately 2x10(6) diversity). Compared with a large nonimmune human scFv library (approximately 1x10(9) diversity), the repertoire of the pseudo-immune scFv library was significantly biased toward the target antigen, which facilitated rapid enrichments of the target-specific high affinity scFvs during selections by fluorescence activated cell sortings. Isolated scFvs, HW5 and HW6, from the pseudo-immune library showed much higher specificity and affinity for the targeted antigen than those from the nonimmune library. Our results suggest that a pseudo-immune antibody library is very efficient to isolate target-specific high affinity antibody from a relatively small sized library.
MeSH terms
HumansImmunoglobulin Variable Region/genetics*Immunoglobulin Variable Region/immunologyImmunoglobulin Variable Region/isolation & purificationImmunoglobulin Variable Region/metabolism*Peptide Library*Receptors, TNF-Related Apoptosis-Inducing LigandReceptors, Tumor Necrosis Factor/immunologySaccharomyces cerevisiae/geneticsSaccharomyces cerevisiae/immunologySaccharomyces cerevisiae/metabolism*Titrimetry
DOI
10.1016/j.bbrc.2006.05.202
PMID
16777066
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Microbiology
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