MONOCLONAL-ANTIBODIES THAT RECOGNIZE FILAMENTOUS PHAGE - TOOLS FOR PHAGE DISPLAY TECHNOLOGY

We generated six hybridoma cell lines that secrete monoclonal antibodi es (mAb) which specifically bind filamentous phage coat proteins. Two of these mAb recognise epitopes that include the N terminus of the coa t protein III (pIII), white 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 ex amples of how these mAb can be exploited in the construction and scree ning of peptide libraries displayed by the filamentous phage major coa t protein. We have used one of these mAb to develop a sensitive ELISA with crude phage supernatants. This assay allows rapid screening of la rge numbers of clones from random peptide phage libraries. Some of the anti-phage mAb described here can interfere with wild-type phage prop agation, 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 p VIII, with respect to vector phage.