The continued development of new tools and techniques that improve sensitivity, selectivity, speed and throughput in biological analysis, while reducing the cost per assay, will be critical in clinical diagnosis, medical research, and other disciplines in the life sciences. Micromachining methods have the potential to enable the construction of low-cost, yet sophisticated microfluidic systems for the analysis of complex protein mixtures. Herein, we propose to develop a phase-changing sacrificial layer (PCSL) microfabrication approach and polymer surface modification methods to create inexpensive microfluidic devices that can improve the detection and quantification of alpha-fetoprotein (AFP), a cancer and fetal wellness marker, in blood and urine. The objectives of this proposal are four-fold. First, we will generalize the PCSL solvent bonding fabrication technique and atom-transfer radical polymerization surface modification methods to a range of polymer microfluidic systems for protein analysis. Second, we plan to evaluate the PCSL solvent bonding approach for making complex, multilayer microfluidic arrays that facilitate the integration of sample processing steps. Third, we will develop these fabrication techniques to enable the incorporation of immunoaffinity purification systems and protein sample preconcentrators for use in enhancing detection in microchip capillary electrophoresis. Fourth, we will utilize the tools from the first three objectives to make microdevices that extract and preconcentrate lower-abundance proteins such as AFP from biological mixtures;we will then evaluate these microsystems for the determination of AFP levels in blood and urine. Importantly, while the studies in this proposal are directed toward the development of miniaturized assays for AFP, these same fabrication and analysis procedures can be generalized in a straightforward manner to allow the rapid quantification of other lower-abundance cancer and disease biomarkers. Thus, PCSL techniques and polymer surface derivatization methods should provide a broadly applicable microchip platform for the electrophoretic analysis and quantification of proteins or other biomolecules in humans.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
5R01EB006124-07
Application #
8212420
Study Section
Enabling Bioanalytical and Biophysical Technologies Study Section (EBT)
Program Officer
Korte, Brenda
Project Start
2006-04-01
Project End
2013-07-31
Budget Start
2012-02-01
Budget End
2013-07-31
Support Year
7
Fiscal Year
2012
Total Cost
$233,139
Indirect Cost
$77,713
Name
Brigham Young University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
009094012
City
Provo
State
UT
Country
United States
Zip Code
84602
Chatterjee, Debolina; Mansfield, Danielle S; Woolley, Adam T (2014) MICROFLUIDIC DEVICES FOR LABEL-FREE AND NON-INSTRUMENTED QUANTITATION OF UNAMPLIFIED NUCLEIC ACIDS BY FLOW DISTANCE MEASUREMENT. Anal Methods 6:8173-8179
Rogers, Chad I; Oxborrow, Joseph B; Anderson, Ryan R et al. (2014) Microfluidic Valves Made From Polymerized Polyethylene Glycol Diacrylate. Sens Actuators B Chem 191:
Kumar, Suresh; Xuan, Jie; Lee, Milton L et al. (2013) Thin-film microfabricated nanofluidic arrays for size-selective protein fractionation. Lab Chip 13:4591-8
Nge, Pamela N; Rogers, Chad I; Woolley, Adam T (2013) Advances in microfluidic materials, functions, integration, and applications. Chem Rev 113:2550-83
Nge, Pamela N; Pagaduan, Jayson V; Yang, Weichun et al. (2013) Integrated affinity and electrophoresis systems for multiplexed biomarker analysis. Methods Mol Biol 919:189-201
Gao, Changlu; Sun, Xiuhua; Woolley, Adam T (2013) Fluorescent measurement of affinity binding between thrombin and its aptamers using on-chip affinity monoliths. J Chromatogr A 1291:92-6
Nge, Pamela N; Yang, Weichun; Pagaduan, Jayson V et al. (2011) Ion-permeable membrane for on-chip preconcentration and separation of cancer marker proteins. Electrophoresis 32:1133-40
Rogers, Chad I; Pagaduan, Jayson V; Nordin, Gregory P et al. (2011) Single-monomer formulation of polymerized polyethylene glycol diacrylate as a nonadsorptive material for microfluidics. Anal Chem 83:6418-25
Yu, Ming; Wang, Qingsong; Patterson, James E et al. (2011) Multilayer polymer microchip capillary array electrophoresis devices with integrated on-chip labeling for high-throughput protein analysis. Anal Chem 83:3541-7
Yang, Weichun; Woolley, Adam T (2010) Integrated Multi-process Microfluidic Systems for Automating Analysis. JALA Charlottesv Va 15:198-209

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