Construction and characterization of a pseudo-immune human antibody library using yeast surface display |
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Authors: | Lee Hae-Won Lee Seung-Hyun Park Kyung-Jin Kim Jeong-Sun Kwon Myung-Hee Kim Yong-Sung |
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Affiliation: | Department of Molecular Science and Technology, Ajou University, San 5, Woncheon-dong, Yeongtong-gu, Suwon 443-749, Republic of Korea. |
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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. |
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Keywords: | Human antibody Pseudo-immune scFv library Nonimmune scFv library Yeast surface display Death receptor 5 |
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