Antigen Dependent Amplification of Phage Antibodies

Antigen specific selection of antibodies (or other proteins) expressed on the surface offilamentous phage has previously been performed by affinity enrichment on columns or by panning. Avaluable addition to this methodology would be to link receptor-ligand binding in a manner similar toclonal proliferation of lymphoid cells during an immune response. Recent studies indicate that it is indeedpossible to link phage display of binding proteins directly to phage replication by genetically fusing theprotein or antigen of interest to capsid protein gene3. Because this approach is unwieldy andinconvenient for most purposes, we will develop a general method to link phage displayed antibodies tophage replication. We will create a chimeric fusion protein linking streptavidin to the terminal 100 aminoacids of the m13 gene3 protein. Next, a test antigen turkey lysozyme will be biotinylated and finally wewill test the capacity of this biotinylated antigen to rescue phage displaying an anti-turkey lysozymescFv. These results will provide the basis for general methods to rapidly generate high affinity antibodiesand other ligands for a diverse range of applications.