Construction and characterization of a pseudo-immune human antibody library using yeast surface display
Construction and characterization of a pseudo-immune human antibody library using yeast surface display
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DOI：
10.1016/j.bbrc.2006.05.202
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发表时间：
2006-08-04
影响因子：
3.1
通讯作者：
Kim, Yong-Sung
中科院分区：
文献类型：
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作者：
Lee, Hae-Won;Lee, Seung-Hyun;Kim, Yong-Sung
在60个健康供体中，有8个个体的淋巴细胞选择了对死亡受体5（DR5）靶抗原的抗体滴度相对较高 - 酵母细胞表面上的链片段变量（SCFV）库（类似于2 x 10（6）多样性）。与大型非免疫人类SCFV文库（类似于1 x 10（9）多样性）相比，伪免疫SCFV库的曲目显着偏向于靶抗原，这有助于促进目标特异性高亲和力SCFV的快速富集通过荧光激活的细胞分数选择。来自伪免疫文库的孤立的SCFV，HW5和HW6比非免疫文库的拟抗原具有更高的特异性和亲和力。我们的结果表明，伪免疫抗体文库非常有效地分离了靶标特异性高亲和抗体，从相对较小的库中分离出来。 （c）2006 Elsevier Inc.保留所有权利。
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 (similar to 2 x 10(6) diversity). Compared with a large nonimmune human scFv library (similar to 1 x 10(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. (c) 2006 Elsevier Inc. All rights reserved.