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(478b) Nanobiosensing Chemicals That Target Nuclear Hormone Receptors: A Rapid, Versatile Cell-Free Protein Synthesis Approach

Authors: 
Bundy, B. C., Brigham Young University
Salehi, A., Brigham Young University
Shakalli Tang, M., The Ohio State University
Yang, S. O., Brigham Young University
Smith, M. T., Brigham Young University
Wood, D. W., The Ohio State University
Hunt, J., Stanford University
There is growing concern of the ubiquitous presence of endocrine disrupting compounds in our environment and their effects on human and animals (monitary impact estimated to be $340 billion USD in the US and $300 billion USD in the EU). This has catalyzed biosensor development to detect and study the interaction of these chemicals with nuclear hormone receptors (NHRs). Although progress has been made, available biosensors remain expensive and time consuming. Here we report a rapid, versatile cell-free protein synthesis platform for detecting chemicals that target NHRs. This biosensor is based on a multidomain-allosteric fusion protein with a core nuclear hormone ligand binding domain and a reporter domain. This biosensor has ability to produce a near real-time signal. We also report the robustness of cell free system in combination with a “just-add-sample” technique to detect endocrine disrupting compounds in environmental samples, including raw sewage. The cell free system has also been shown to be functional in human samples including blood and urine, which expands the possible portable applications of the biosensor.

Citation:

Salehi ASM, Shakali Tang MJ, Smith MT, Hunt JM, Law RA, Wood DW, Bundy BC. Cell-Free Protein Synthesis Approach to Biosensing hTRβ-Specific Endocrine Disruptors. Analytical Chemistry. 2017. 89(6):3395–3401