The powerful combination of phage surface display of cDNA libraries and high throughput screening

Phage surface display of cDNA libraries facilitates cloning, expression and rapid selection of functional gene products physically linked to their gen etic information through gene product-ligand interactions. Efficient screen ing technologies based on selective enrichment of clones expressing desired gene products allows, within a short time, the isolation of all ligand-spe cific clones that are present in a library. Manual identification of clones by restriction analysis and random sequencing is unlike to be successful f or the isolation of gene products derived from rare mRNA species resulting from selection of the libraries using polyvalent ligands like serum from pa tients. Here we describe rapid handling of large numbers of individual clon es selected from molecular libraries displayed on phage surface using the p ower of robotics-based high throughput screening. The potential of the comb ination of cDNA-phage surface display, with selection for specific interact ions by functional screening and robotic technology is illustrated by the i solation of more sequences potentially encoding IgE-binding proteins than p ostulated from Western blot analyses using extracts derived from raw materi al of complex allergenic sources. The subsequent application of functional enrichment and robotics-based screening will facilitate the rapid generatio n of information about the repertoire of protein structures involved in all ergic diseases.