Mazor Yariv, Van Blarcom Thomas, Mabry Robert, Iverson Brent L, Georgiou George
Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, Texas 78712, USA.
Nat Biotechnol. 2007 May;25(5):563-5. doi: 10.1038/nbt1296. Epub 2007 Apr 15.
We describe facile isolation of full-length IgG antibodies from combinatorial libraries expressed in E. coli. Full-length heavy and light chains are secreted into the periplasm, where they assemble into aglycosylated IgGs that are captured by an Fc-binding protein that is tethered to the inner membrane. After permeabilizing the outer membrane, spheroplast clones expressing so-called E-clonal antibodies, which specifically recognize fluorescently labeled antigen, are selected using flow cytometry. Screening of a library constructed from an immunized animal yielded several antibodies with nanomolar affinities toward the protective antigen of Bacillus anthracis.
我们描述了从大肠杆菌中表达的组合文库中轻松分离全长IgG抗体的方法。全长重链和轻链分泌到周质中，在那里它们组装成无糖基化的IgG，被连接到内膜的Fc结合蛋白捕获。在使外膜通透后，使用流式细胞术选择表达所谓E克隆抗体（特异性识别荧光标记抗原）的原生质球克隆。对来自免疫动物构建的文库进行筛选，得到了几种对炭疽芽孢杆菌保护性抗原有纳摩尔亲和力的抗体。
