Multiplex ELISA in a single microfluidic channel |
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Authors: | Yanagisawa Naoki Mecham James O Corcoran Robert C Dutta Debashis |
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Institution: | (1) Department of Chemistry (Dept. # 3838), University of Wyoming, 1000 East University Avenue, Laramie, WY 82071, USA;(2) USDA-ARS, Arthropod Borne Animal Disease Research Laboratory, Laramie, WY 82071, USA; |
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Abstract: | In this article, we demonstrate a novel approach to implementing multiplex enzyme-linked immunosorbent assay (ELISA) in a
single microfluidic channel by exploiting the slow diffusion of the soluble enzyme reaction product across the different assay
segments. The functionality of the reported device is realized by creating an array of ELISA regions within a straight conduit
that are selectively patterned with chosen antibodies/antigens via a flow-based method. The different analytes are then captured
in their respective assay segments by incubating a 5-μL aliquot of sample in the analysis channel for an hour under flow conditions.
Once the ELISA surfaces have been prepared and the enzyme substrate introduced into the analysis channel, it is observed that
the concentration of the soluble enzyme reaction product (resorufin) at the center of each assay region grows linearly with
time. Further, the rate of resorufin generation at these locations is found to be proportional to the concentration of the
analyte being assayed in that segment provided that the ELISA reaction time in the system (τ
R
) is kept much shorter than that required by the resorufin molecules to diffuse across an assay segment (τ
D
). Under the operating condition τ
R
<< τ
D
, the reported device has been shown to have a 35% lower limit of detection for the target analyte concentration compared
with that on a commercial microtiter plate using only a twentieth of the sample volume. |
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Keywords: | |
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