Lee Hae-Won, Lee Seung-Hyun, Park Kyung-Jin, Kim Jeong-Sun, Kwon Myung-Hee, Kim Yong-Sung
Department of Molecular Science and Technology, Ajou University, San 5, Woncheon-dong, Yeongtong-gu, Suwon 443-749, Republic of Korea.
Biochem Biophys Res Commun. 2006 Aug 4;346(3):896-903. doi: 10.1016/j.bbrc.2006.05.202. Epub 2006 Jun 9.
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.
在60名健康供体中，挑选出8名血浆对死亡受体5（DR5）的一种靶抗原显示出相对较高抗体滴度的个体的淋巴细胞，作为抗体基因来源，在酵母细胞表面构建所谓的抗DR5假免疫人单链可变片段（scFv）文库（多样性约为2×10⁶）。与一个大型非免疫人scFv文库（多样性约为1×10⁹）相比，假免疫scFv文库的库显著偏向于靶抗原，这有助于在通过荧光激活细胞分选进行筛选期间快速富集靶标特异性高亲和力scFv。从假免疫文库中分离出的scFv，HW5和HW6，对靶向抗原显示出比非免疫文库中的scFv更高的特异性和亲和力。我们的结果表明，假免疫抗体文库对于从相对较小规模的文库中分离靶标特异性高亲和力抗体非常有效。
