Isolation of SFV's That Regulate Transcription

The development of non-animal based methods of antibody production has become increasingly favoredyears. In addition, the development of methods of producing human-derived or humanized versions of mantibodies has paralleled the attempts to develop the non-animal systems. Concurrently, other groupsstudying primarily signal transduction pathways and/or transcriptional regulation have developed sevassessing the protein/protein interactions inherent in these systems. In this application, we will cadvances and existing needs of both of the aforementioned groups to produce a genetic selection systbased on a transcriptional readout assay of reporter gene function with the goal of isolating singleimmunoglobulin variable domains (sFv's) with high affinity for desired antigens. The system is basedsystem described originally by Fields and Song (Nature 340:245, 1989). A library of human-derived sFinto vectors that will encode fusion proteins linking these sFv's with constitutive transactivationlocalization signals. The goal of these initial studies will be to isolate a fusion molecule from thtranscriptional activation domain to a specific DNA-bound factor.