New molecular information from blood is being used to improve patient outcomes. Assays for established biomarkers have recently advanced (so-called 'high sensitivity', 'variants'and 'ultra', for examples), allowing physicians to use this additional information to provide better care. The enhanced patient outcomes result from more detailed, accurate and precise diagnostic and risk stratification, resulting in more timely and useful care (such as pharmaceuticals, treatments options, and timing of treatments) and avoiding unnecessary and costly actions. The objective of this proposal is to enable a new capability to isolate and concentrate biomarkers from blood in small volumes (200 microliters), at high sensitivity, over short periods of time, and to monitor multiple markers. We initially will focus on biomarkers for myocardial infarction and stroke. Upon completion of the entire project-which this proposal enables-enhanced information will be provided to physicians, allowing for accurate and fast diagnostics and treatments providing better patient outcomes-saving lives and money. To accomplish the proposal objective, a pair of microfluidic techniques will allow processing of small samples of blood to remove unwanted materials and isolate and concentrate the target biomarkers. These two techniques enable the objective because they keep the sample volume minimal and remove unwanted materials that could degrade detection, while quickly isolating and concentrating target species. Physically, this is made possible by exploit a unique combination of dielectrophoretic, flow and electrophoretic forces combined into the two techniques (gradient dielectrophoresis and electrophoretic capture) pioneered in the PI's laboratory. Gradient dielectrophoresis will remove cells and debris (and perhaps concentrate the targets) and electrophoretic capture will isolate and concentrate individual biomarkers away from possible interfering species. The four target biomarkers identified for proof of principle to enable the larger project are: two cardiac markers- myoglobin and cardiac troponin I, (cTnI), a stroke marker-neuron specific enolase (NSE), and an inflammatory marker-tumor necrosis factor-alpha (TNF1). These targets will provide a reasonable test for the success of the strategy and techniques..
Developing an ability to isolate and concentrate biomarkers from blood using gradient dielectrophoresis and electrophoretic capture improving detection limits by at least two orders of magnitude while keeping the sample volume very modest (200 microliters).
|Woolley, Christine F; Hayes, Mark A (2015) Sensitive Detection of Cardiac Biomarkers Using a Magnetic Microbead Immunoassay. Anal Methods 7:8632-8639|
|Woolley, Christine F; Hayes, Mark A; Mahanti, Prasun et al. (2015) Theoretical limitations of quantification for noncompetitive sandwich immunoassays. Anal Bioanal Chem 407:8605-15|
|Kenyon, Stacy M; Keebaugh, Michael W; Hayes, Mark A (2014) Development of the resolution theory for electrophoretic exclusion. Electrophoresis 35:2551-9|
|Jones, Paul V; DeMichele, Alexa F; Kemp, LaKeta et al. (2014) Differentiation of Escherichia coli serotypes using DC gradient insulator dielectrophoresis. Anal Bioanal Chem 406:183-92|
|Staton, Sarah J R; Castillo, Josemar A; Taylor, Thomas J et al. (2013) Identifying indoor environmental patterns from bioaerosol material using HPLC. Anal Bioanal Chem 405:351-7|
|Woolley, Christine F; Hayes, Mark A (2013) Recent developments in emerging microimmunoassays. Bioanalysis 5:245-64|
|Yanashima, Ryan; García, Antonio A; Aldridge, James et al. (2012) Cutting a drop of water pinned by wire loops using a superhydrophobic surface and knife. PLoS One 7:e45893|
|Staton, Sarah J R; Jones, Paul V; Ku, Ginger et al. (2012) Manipulation and capture of Aýý amyloid fibrils and monomers by DC insulator gradient dielectrophoresis (DC-iGDEP). Analyst 137:3227-9|
|Castillo, Josemar A; Staton, Sarah J R; Taylor, Thomas J et al. (2012) Exploring the feasibility of bioaerosol analysis as a novel fingerprinting technique. Anal Bioanal Chem 403:15-26|
|Kenyon, Stacy M; Weiss, Noah G; Hayes, Mark A (2012) Using electrophoretic exclusion to manipulate small molecules and particles on a microdevice. Electrophoresis 33:1227-35|
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