Efficient recovery of high-affinity antibodies from a single-chain Fab yeast display library.
Efficient recovery of high-affinity antibodies from a single-chain Fab yeast display library.
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DOI：
10.1016/j.jmb.2009.04.019
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发表时间：
2009-06-05
影响因子：
5.6
通讯作者：
Burton DR
中科院分区：
文献类型：
--
作者：
Walker LM;Bowley DR;Burton DR
酵母展示技术是一种分离针对目标抗原的单克隆抗体（mab）的有效技术。抗体文库在酵母表面显示为单链可变片段（scFv）和抗原结合片段（Fab）。在这里，我们将这两种格式结合起来，在酵母表面以单链fab （scFab）的形式展示了具有良好特征的单抗，并构建了第一个scFab酵母展示抗体库。当在酵母表面表达时，三种抗人类免疫缺陷病毒(HIV)-1单克隆抗体中有两种与scFvs结合的亲和力更高。此外，可溶的scFab制剂表现出与相应的Fab片段相当的结合和中和特征。在酵母表面展示免疫HIV-1 scFab文库，随后进行针对HIV-1 gp120的分选，允许选择13个抗原特异性克隆。当使用相同的cDNA构建scFv格式文库时，选择了数量相近但亲和力较低的一组克隆。基于这些结果，可以高效地构建和选择酵母显示的scFab文库，无需重新格式化步骤，并用于分离比scFv文库高亲和力的抗体。
Yeast display is a powerful technology for the isolation of monoclonal antibodies (mAbs) against a target antigen. Antibody libraries have been displayed on the surface of yeast as both single-chain variable fragment (scFv) and antigen binding fragment (Fab). Here, we combine these two formats to display well-characterized mAbs as single-chain Fabs (scFabs) on the surface of yeast and construct the first scFab yeast display antibody library. When expressed on the surface of yeast, two out of three anti-human immunodeficiency virus (HIV)-1 mAbs bound with higher affinity as scFabs than scFvs. Also, the soluble scFab preparations exhibited binding and neutralization profiles comparable to that of the corresponding Fab fragments. Display of an immune HIV-1 scFab library on the surface of yeast, followed by rounds of sorting against HIV-1 gp120, allowed for the selection of 13 antigen-specific clones. When the same cDNA was used to construct the library in an scFv format, a similar number but a lower affinity set of clones were selected. Based on these results, yeast-displayed scFab libraries can be constructed and selected with high efficiency, characterized without the need for a reformatting step, and used to isolate higher-affinity antibodies than scFv libraries.
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影响因子：
64.5
作者：
Choe, H;Li, WH;Farzan, M
通讯作者：
Farzan, M
DOI：
10.1016/j.bbrc.2006.05.202
发表时间：
2006-08-04
影响因子：
3.1
作者：
Lee, Hae-Won;Lee, Seung-Hyun;Kim, Yong-Sung
通讯作者：
Kim, Yong-Sung
在巨大芽孢杆菌中生产单链 Fab (scFab) 片段。
影响因子：
6.4
作者：
Jordan, Eva;Al-Halabi, Laila;Schirrmann, Thomas;Hust, Michael;Duebel, Stefan
通讯作者：
Duebel, Stefan
影响因子：
1.6
作者：
Kirschner, Marc;Monrose, Val;Moore, John P.
通讯作者：
Moore, John P.
影响因子：
1.5
作者：
BUCHACHER, A;PREDL, R;KATINGER, H
通讯作者：
KATINGER, H