The post-genomics era will demand high throughput technologies to generate and evolve proteins and protein-binding molecules. The overall goal of this work is to develop a novel ribo-display technology for in vitro molecular evolution. Preliminary work provides proof-of-concept. The next step is to demonstrate the utility and robustness of this novel display system. To attain this goal we will prepare large, highly diverse human scFv libraries and utilize these libraries to identify high affinity antibodies. In phase I we will amplify human antibody sequences to generate a large scFv library and construct ribodisplay vectors. In vitro translation followed by biopanning will be used to identify specific antigen binding scFvs. These will be expressed, purified and characterized by binding and affinity measurements. This simple yet high-yield system should contribute significantly to proteomics research in the post-genome era.
