The protein distribution in the blood can indicate the overall state or condition of the human body. Therefore, determination of the proteins in blood samples can serve as a powerful diagnostic tool for early detection of disease states, such as cancer. Unfortunately, there is no analytical method available today to provide rapid high resolution profiles of proteins present in complex mixtures such as blood. The overall objective of this proposal is to develop a new method called electromobility focusing (EMF) for separating, concentrating, and identifying proteins in complex mixtures with unsurpassed resolution. The physical description of the device for EMF is a channel containing a buffer solution along which a high voltage is applied, similar to capillary electrophoresis, except that the electric field intensity along the column is not linear, but is a continuous gradient. Proteins can be separated and focused with exceptionally high resolution in the channel by applying a pressure-induced liquid flow counter to the electrophoretic migration direction of the proteins. The resulting forces acting on the proteins concentrate them into narrow bands at specific positions along the channel in order of their electrophoretic mobilities. The proteins can be mobilized in the separation channel by changing the magnitude of the pressure-induced counter flow or by reducing the applied voltage. The specific objectives of this work include the development of two different device formats for performing EMF: (a) semi-preparative and (b) microchip. EMF will be applied to the rapid quantitative detection of five known protein tumor markers in blood serum, including prostate specific antigen, carcinoembryonic antigen, carcinoma-associated antigen 1 25, alpha-fetoprotein, and humanchorionic gonadotropin. Proposed new tumor marker, thymidine kinase I, will also be studied.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM064547-02
Application #
6698842
Study Section
Special Emphasis Panel (ZRG1-SSS-6 (10))
Program Officer
Edmonds, Charles G
Project Start
2003-02-01
Project End
2007-01-31
Budget Start
2004-02-01
Budget End
2005-01-31
Support Year
2
Fiscal Year
2004
Total Cost
$368,939
Indirect Cost
Name
Brigham Young University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
009094012
City
Provo
State
UT
Country
United States
Zip Code
84602
Heywood, Matthew S; Farnsworth, Paul B (2010) Optimization of native fluorescence detection of proteins using a pulsed nanolaser excitation source. Appl Spectrosc 64:1283-8
Sun, Xuefei; Li, Dan; Woolley, Adam T et al. (2009) Bilinear electric field gradient focusing. J Chromatogr A 1216:6532-8
Sun, Xuefei; Farnsworth, Paul B; Tolley, H Dennis et al. (2009) Performance optimization in electric field gradient focusing. J Chromatogr A 1216:159-64
Sun, Xuefei; Li, Dan; Lee, Milton L (2009) Poly(ethylene glycol)-functionalized polymeric microchips for capillary electrophoresis. Anal Chem 81:6278-84
Sun, Xuefei; Liu, Jikun; Lee, Milton L (2008) Surface modification of polymer microfluidic devices using in-channel atom transfer radical polymerization. Electrophoresis 29:2760-7
Sun, Xuefei; Farnsworth, Paul B; Woolley, Adam T et al. (2008) Poly(ethylene glycol)-functionalized devices for electric field gradient focusing. Anal Chem 80:451-60
Sun, Xuefei; Liu, Jikun; Lee, Milton L (2008) Surface modification of glycidyl-containing poly(methyl methacrylate) microchips using surface-initiated atom-transfer radical polymerization. Anal Chem 80:856-63
Lin, Shu-Ling; Li, Yuanyuan; Woolley, Adam T et al. (2008) Programed elution and peak profiles in electric field gradient focusing. Electrophoresis 29:1058-66
Gu, Binghe; Li, Yun; Lee, Milton L (2007) Polymer monoliths with low hydrophobicity for strong cation-exchange capillary liquid chromatography of peptides and proteins. Anal Chem 79:5848-55
Pan, Tao; Fiorini, Gina S; Chiu, Daniel T et al. (2007) In-channel atom-transfer radical polymerization of thermoset polyester microfluidic devices for bioanalytical applications. Electrophoresis 28:2904-11

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