title: Construction and characterization of a pseudo-immune human antibody library using yeast surface display.
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.
authors.affiliation: Department of Molecular Science and Technology, Ajou University, San 5, Woncheon-dong, Yeongtong-gu, Suwon 443-749, Republic of Korea.
authors.forename: Hae-Won
authors.initials: HW
authors.lastname: Lee
authors.forename: Seung-Hyun
authors.initials: SH
authors.lastname: Lee
authors.forename: Kyung-Jin
authors.initials: KJ
authors.lastname: Park
authors.forename: Jeong-Sun
authors.initials: JS
authors.lastname: Kim
authors.forename: Myung-Hee
authors.initials: MH
authors.lastname: Kwon
authors.forename: Yong-Sung
authors.initials: YS
authors.lastname: Kim
mesh_terms: D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D019151:Peptide Library; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand; D018124:Receptors, Tumor Necrosis Factor / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D017186:Titrimetry
chemical_list: D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand; D018124:Receptors, Tumor Necrosis Factor; C505639:TNFRSF10B protein, human