Dente L, Cesareni G, Micheli G, Felici F, Folgori A, Luzzago A, Monaci P, Nicosia A, Delmastro P
Dipartimento di Biologia, Università di Roma Tor Vergata, Italy.
Gene. 1994 Oct 11;148(1):7-13. doi: 10.1016/0378-1119(94)90227-5.
We generated six hybridoma cell lines that secrete monoclonal antibodies (mAb) which specifically bind filamentous phage coat proteins. Two of these mAb recognise epitopes that include the N terminus of the coat protein III (pIII), while two others are specific for the N terminus of the major coat protein VIII (pVIII). These mAb are valuable tools to study phage assembly and structure. Furthermore, we describe two examples of how these mAb can be exploited in the construction and screening of peptide libraries displayed by the filamentous phase major coat protein. We have used one of these mAb to develop a sensitive ELISA with crude phage supernatants. This assay allows rapid screening of large numbers of clones from random peptide phage libraries. Some of the anti-phage mAb described here can interfere with wild-type phage propagation, while phage carrying modifications in their coat proteins are insensitive to growth inhibition. We have exploited this observation as a tool to favour the growth of phage displaying peptides fused to pVIII, with respect to vector phage.
我们生成了六种杂交瘤细胞系,它们分泌能特异性结合丝状噬菌体外壳蛋白的单克隆抗体(mAb)。其中两种mAb识别的表位包括外壳蛋白III(pIII)的N端,另外两种则对主要外壳蛋白VIII(pVIII)的N端具有特异性。这些mAb是研究噬菌体组装和结构的宝贵工具。此外,我们描述了两个实例,说明如何利用这些mAb构建和筛选由丝状噬菌体主要外壳蛋白展示的肽库。我们已使用其中一种mAb开发了一种用于粗噬菌体上清液的灵敏ELISA。该检测方法可快速筛选来自随机肽噬菌体文库的大量克隆。本文所述的一些抗噬菌体mAb可干扰野生型噬菌体的繁殖,而外壳蛋白带有修饰的噬菌体对生长抑制不敏感。我们利用这一观察结果作为一种工具,相对于载体噬菌体而言,促进展示与pVIII融合肽的噬菌体的生长。