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Proof-of-concept study for the development of a cell-free GPCR-based biosensor for rapid non-specific detection of chemical and biological toxic agents

Award No.: 
2007-SY-061-000003
Status: 
Complete
Principal Investigator: 
Evangelyn Alocilja
PI Organization: 
Michigan State University
Abstract: 
This project is a high-risk/high-reward exploratory proof-of-concept work to evaluate the potential of G-protein coupled receptors (GPCRs) as sensing elements in biosensor devices for rapid non-specific but highly sensitive detection (detect-to-warn) of toxicants in selected complex food matrices. G-protein coupled receptors (GPCRs) constitute a large and diverse family of proteins whose primary function is to initiate a variety of cellular responses to a diverse array of extracellular stimuli. The diversity of GPCRs is dictated both by the multiplicity of stimuli to which they respond, as well as by the variety of intracellular signaling pathways they activate. Recently, there is a high interest in the study and understanding of GPCRs for drug development and evaluation. At the moment, there is hardly any biosensor based on GPCRs. In this exploratory work, we will use acetylcholine receptor (AChR) as the biological sensing element in an electrochemical biosensor. AChR is normally present in animals and acts as a mediator of neural transmission. Such a biosensor will be useful for the determination of organophosphorus compounds, drugs, naturally-occurring toxins, and a variety of environmental chemicals and pollutants. We will then evaluate its electrical (conductivity/resistivity) and electrochemical (reductrion/oxidation) responses to the presence of various types of toxicants. We will also evaluate the contributory effect of the food matrix on the detection signal. For this exploratory work, we will evaluate milk and eggs because they are frequently consumed by the very young, a highly vulnerable group of the population, as well as they are a component of many downstream nutritious products. Any malicious contamination in these food products can have catastrophic consequences on public health, food supply, and economic infrastructure. This exploratory work will pave the way for the future development of a cell-free detection scheme that will satisfy the following performance criteria: detect-to-warn in less than 15 min, inexpensive (