Løset G Å, Lunde E, Bogen B, Brekke O H, Sandlie I
Department of Molecular Biosciences, University of Oslo, N-0316 Oslo, Norway.
Protein Eng Des Sel. 2007 Sep;20(9):461-72. doi: 10.1093/protein/gzm044. Epub 2007 Oct 9.
Phage display has been instrumental for the success of antibody (Ab) technology. The aim of the present study was to explore phage display of soluble T-cell receptors (TCRs). A library platform that supports engineering and selection of improved TCRs to be used as detection reagents for specific antigen presentation will be very useful. In such applications, high, equal and clone independent display levels are a prerequisite for 'fair' selection. Therefore, we explored how different pIII fusion formats and modes affected the display levels of two murine alpha/beta TCRs. Both are derived from T-cell clones associated with the MOPC315 myeloma model. The results show that the design of the pIII fusion particle significantly affects the subsequent display levels. Furthermore, successful display may be obtained both in phagemid and phage versions. Importantly, improvement of poor display can be achieved by over-expressing the periplasmic chaperone FkpA.
噬菌体展示技术对抗体（Ab）技术的成功起到了重要作用。本研究的目的是探索可溶性T细胞受体（TCR）的噬菌体展示。一个支持工程改造和筛选改良型TCR以用作特定抗原呈递检测试剂的文库平台将非常有用。在这类应用中，高、均等且不依赖克隆的展示水平是“公平”筛选的前提条件。因此，我们探究了不同的pIII融合形式和模式如何影响两种小鼠α/β TCR的展示水平。这两种TCR均来源于与MOPC315骨髓瘤模型相关的T细胞克隆。结果表明，pIII融合颗粒的设计显著影响后续的展示水平。此外，在噬菌粒和噬菌体形式中均可成功实现展示。重要的是，通过过表达周质伴侣蛋白FkpA可以改善较差的展示效果。
