TY - JOUR
T1 - Dual-column miniseparator to separate alpha-fetoprotein from fibrinogen and immunoglobulin
AU - Yang, Yongmo
AU - Chae, Junseok
N1 - Funding Information:
Manuscript received November 6, 2009; revised May 10, 2010; accepted May 23, 2010. Date of publication July 19, 2010; date of current version October 1, 2010. This work was supported in part by the National Science Foundation under Grant ECCS-0901440. Subject Editor A. J. Ricco. The authors are with Arizona State University, Tempe, AZ 85287 USA (e-mail: yongmo.yang@asu.edu). Color versions of one or more of the figures in this paper are available online at http://ieeexplore.ieee.org. Digital Object Identifier 10.1109/JMEMS.2010.2055542
PY - 2010/10
Y1 - 2010/10
N2 - This paper reports a dual-column miniseparator that depletes and separates target analytes from a mixed sample. The prototype was designed to address the need for a sample preparation stage in portable point-of-care testing devices, and its efficacy was tested with Immunoglobulin G (IgG), α-fetoProtein (AFP), and fibrinogen. The miniseparator connects two minicolumns in series: The first column is an affinity minicolumn containing Protein A/G agarose beads, and the second column is a size exclusion chromatography (SEC) minicolumn that separates mixed analytes by size. When a mixture of IgG, fibrinogen, and AFP flows through the dual-column miniseparator, IgG is depleted in the first column, and AFP is separated from fibrinogen in the second column. Each protein was conjugated with fluorescent dyes, and IgG depletion and SEC separation results were confirmed by evaluating the eluents' fluorescent intensity. Red (IgG), green (AFP), and blue (fibrinogen) intensities showed that, in 10 min, the first column (affinity minicolumn) depleted over 99.5% of IgG, and the second column (SEC minicolumn) separated AFP (1 μg/mL} ) from a fibrinogen-rich solution (10 μg/mL}) with 1.76 resolution and 0.093-mm plate height.
AB - This paper reports a dual-column miniseparator that depletes and separates target analytes from a mixed sample. The prototype was designed to address the need for a sample preparation stage in portable point-of-care testing devices, and its efficacy was tested with Immunoglobulin G (IgG), α-fetoProtein (AFP), and fibrinogen. The miniseparator connects two minicolumns in series: The first column is an affinity minicolumn containing Protein A/G agarose beads, and the second column is a size exclusion chromatography (SEC) minicolumn that separates mixed analytes by size. When a mixture of IgG, fibrinogen, and AFP flows through the dual-column miniseparator, IgG is depleted in the first column, and AFP is separated from fibrinogen in the second column. Each protein was conjugated with fluorescent dyes, and IgG depletion and SEC separation results were confirmed by evaluating the eluents' fluorescent intensity. Red (IgG), green (AFP), and blue (fibrinogen) intensities showed that, in 10 min, the first column (affinity minicolumn) depleted over 99.5% of IgG, and the second column (SEC minicolumn) separated AFP (1 μg/mL} ) from a fibrinogen-rich solution (10 μg/mL}) with 1.76 resolution and 0.093-mm plate height.
KW - Cancer biomarker
KW - point of care (POC)
KW - size exclusion chromatography (SEC)
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U2 - 10.1109/JMEMS.2010.2055542
DO - 10.1109/JMEMS.2010.2055542
M3 - Article
AN - SCOPUS:77957578100
SN - 1057-7157
VL - 19
SP - 1041
EP - 1049
JO - Journal of Microelectromechanical Systems
JF - Journal of Microelectromechanical Systems
IS - 5
M1 - 5512546
ER -