Abstract: Provided herein are methods and systems allowing for real-time measurement of cellular drug-target engagement. Use of fluorescence-based cell target engagement technology with real-time gene expression machinery provides several advantages over prior systems. Integration with real-time gene expression machinery without bias to any particular design or brand is provided. Programmability of cell target engagement methodology, such that any software in real-time gene expression machine can seamlessly be used for programming is provided. Single or multiple machine integration for the use of cell target engagement technology is provided. Compatibility with multiple multi-well plate setups is provided. Development of unique modifications of real-time (quantitative) gene expression software to detect real-time cellular drug-target engagement to work efficiently with existing gene expression machinery is provided.

Abstract: Provided herein are methods amenable to high-throughput multiplexing, in part, using a modified enzyme complementation assay, that can be used to screen a library of test compounds and to identify compounds that inhibit denaturation of a target polypeptide of interest.