Abstract: Disclosed is a method that combines high throughput and flexible nature of a cell-free protein microarray with the quantitative capability of surface plasmon resonance to detect >400 different protein interactions in <1 hour. A method of detecting interactions between a targeting agent and one or more proteins of interest is disclosed. The method includes producing a set of proteins of interest using a cell-free protein expression system; providing the set of proteins of interest on a protein microarray wherein each spot in the array comprises a protein of interest; contacting the protein microarray with a targeting agent that binds to one or more of the set of proteins of interest; and detecting the binding of the targeting agent to the set of proteins of interest using surface plasmon resonance imaging (SPRi), thereby detecting the targeting agent and one or more proteins of interest in the micro array.

Abstract: Cell lines harboring a range of polymorphisms using recombinant cytochrome P450 or other chemical-metabolizing enzymes in a parent cell line that is minimally expressing or devoid of its own cytochrome P450 protein or other chemical-metabolizing enzymes of interest can be placed into an array format to enable high throughput screening of one or more chemicals for CYP450 or other enzyme-dependent metabolism (FIG. 9). Processing of the cells can be automated, done en-masse through use of an array having a substrate upon which a plurality of cell lines with exogenous chemical-metabolizing enzymes are coupled, and both relative and quantitative metabolism rates determined using mass spectrometry over time. Thus, methods are disclosed to measure, for example, the effects of cytochrome P450 polymorphisms on clinical drug metabolism in a high throughput manner for drug development and genetically personalized diagnostics and treatment regimens.