TY - JOUR
T1 - Development and characterization of an ELISA assay in PDMS microfluidic channels
AU - Eteshola, E.
AU - Leckband, D.
N1 - Funding Information:
The work described in this paper was supported by the Defense Advanced Research Projects (DARPA) F33615-98-1-2853. The fluorescence measurements were carried out at the Fluorescence Dynamics Laboratory (FDL — a national resource laboratory supported by the NIH), Department of Physics, and University of Illinois at Urbana-Champaign. We thank Kathy Motsegood for her excellent technical assistance with the mask fabrication.
PY - 2001/1/25
Y1 - 2001/1/25
N2 - This report describes the first preliminary evaluation of a heterogeneous sandwich enzyme-linked immunoassay in a PDMS microfluidic device. The PDMS devices were fabricated using replica molding against a patterned photoresist generated by photolithographic techniques. With this experimental setup, the microfluidic sensor chip was successfully used to quantify a model analyte (sheep IgM) with sensitivity down to 17 nM. The conventional blocking cocktail used in nearly all polystyrene microtiter plate-based ELISA assays failed to block the nonspecific adsorption of the analyte and secondary antibody. This report describes the successful use of surface modification chemistries and a modified blocking cocktail to reduce background due to nonspecific adsorption and to thereby increase the sensitivity of the assay. These protocols can be extended to the detection of a variety of analytes by immunoassay in PDMS microchannels.
AB - This report describes the first preliminary evaluation of a heterogeneous sandwich enzyme-linked immunoassay in a PDMS microfluidic device. The PDMS devices were fabricated using replica molding against a patterned photoresist generated by photolithographic techniques. With this experimental setup, the microfluidic sensor chip was successfully used to quantify a model analyte (sheep IgM) with sensitivity down to 17 nM. The conventional blocking cocktail used in nearly all polystyrene microtiter plate-based ELISA assays failed to block the nonspecific adsorption of the analyte and secondary antibody. This report describes the successful use of surface modification chemistries and a modified blocking cocktail to reduce background due to nonspecific adsorption and to thereby increase the sensitivity of the assay. These protocols can be extended to the detection of a variety of analytes by immunoassay in PDMS microchannels.
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U2 - 10.1016/S0925-4005(00)00640-7
DO - 10.1016/S0925-4005(00)00640-7
M3 - Article
AN - SCOPUS:0035152676
VL - 72
SP - 129
EP - 133
JO - Sensors and Actuators B: Chemical
JF - Sensors and Actuators B: Chemical
SN - 0925-4005
IS - 2
ER -