Abstract

Chromobodies and the Fluorescent 2-Hybrid Assay: Real Time Visualization of Proteins and Protein-protein Interactions for High Content Analysis

Ulrich Rothbauer, Professor, Natural and Medical Sciences Institute at the University of Tuebingen

One of the major challenges in early drug discovery is to understand cellular processes in response to external stimuli. The main goal is to localize cellular components and understand their interactions. To visualize dynamic changes in real time, we have developed a new format of intracellular functional antibodies (chromobodies). In combination with high-throughput microscopy and automated image analysis we have demonstrated that chromobodies can target and trace endogenous cellular components for high-content analysis (HCA) in living cells. Recent applications of new chromobody based cellular models are used to monitor cell cycle, dynamic rearrangement of the cytoskeleton upon compound treatment and signal transduction within living cells.To follow reversible protein-protein interactions in living cells we have developed the Fluorescent 2-Hybrid (F2H) technology. By tethering a fluorescent bait at a defined cellular structure, we generate a positional protein-protein interaction platform and assay for co-localization of fluorescent prey proteins in real time. The results of case studies which demonstrate the potential of the F2H assay for compound screening will be reported.